BASIC AND TRANSLATIONAL MEDICINE
BASIC RESEARCH OF CARDIOVASCULAR DISEASE
GW31-e0016
Cong Fu1,2, Qiancheng Xu3, Shengxing Tang1, Yuhan Cao2,4
1Department of Cardiology, Yi Ji Shan Hospital Affiliated to Wan Nan Medical College
2Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institution, Wan Nan Medical College
3Department of Critical Care Medicine, Yi Ji Shan Hospital Affiliated to Wan Nan Medical College
4Department of Nephrology, Yi Ji Shan Hospital Affiliated to Wan Nan Medical College
OBJECTIVES Triggering receptor expressed on myeloid cells-2 (TREM-2) is a cell surface receptor primarily expressed on monocyte-derived cells. Recent years, the organ protecting effect of TREM-2 attracted researchers’ attention. However, the role of TREM-2 in other system especially in circulating was still not well demonstrated. If TREM-2 have cardiac protecting effect needed to be further studied. Accordingly, we designed this research to determine the role of TREM-2 in myocardial infarction (MI) and preliminary demonstrated the molecular mechanism.
METHODS Recombinant adenovirus containing the gene coding full-length mouse TREM-2 and EGFP and control vector containing EGFP gene without any transgene were produced using the ViraPower Adenoviral Expression System. The male C57BL6 mice (weight 20–22 g) were anesthetized with intraperitoneal pentobarbital (35 mg/kg) and intubated. The left anterior descending (LAD) coronary artery was ligated proximally with 7-0 silk suture via a left thoracotomy incision. After LAD ligated, animals were randomly divided into three groups: (1) Mice were subjected to LAD coronary ligation; (2) Mice were received 50 μL of intramyocardial injections of 5*105 Ad.Null; (3) Mice were received 50 μL of intramyocardial injections of 5*105 Ad. TREM-2. The injection point was above ligation site. Non-operated control and sham surgery were performed. After 7 days, 2-dimensional echocardiography was performed on the mice using a transthoracic echocardiogram to measure the cardiac function. All the animals were executed by over-dose of intraperitoneal pentobarbital (130 mg/kg) after echocardiography. The expression of GFP and Trem-2 were detected. HE staining, Masson staining and TUNEL staining were performed to detect the pathological changes, infarcted size and cardiomyocytes apoptosis. Western blot was performed to measure the expression of signal protein in myocardial tissue.
RESULTS GFP detection showed that both Ad. TREM-2 and Ad.Null successfully transfected in cardiomyocytes. TREM-2 expressed in cardiomyocytes after Ad. TREM-2 transfection was higher compared to Ad. Null transfection. Cardiac function was evaluated at 10 days after MI. MI group had lower LVEF and LVFS than control and Sham. The LVEF and LVFS were significantly higher in Ad. TREM-2 group than MI and Ad. Null group. Further, the LVIDd and LVIDs were higher in MI group than control and sham. Ad. TREM-2 group had lower LVIDd and LVIDs than MI and Ad.Null. HE staining showed inflammatory cell infiltration, myocardial dissolution and cardiomyocyte apoptosis in MI group compared to control and sham. In the Ad. TREM-2 group, the myocardial injury was significantly alleviated compared to MI and Ad. Nul. Masson staining showed remarkably reduced infarct size in the Ad. TREM-2 group compared to MI and Ad. Null. TUNEL staining showed that apoptotic cardiomyocytes increased markedly in the myocardium tissue in the MI group compared to control and sham. Ad. TREM-2 transfection decreased the number of apoptotic cells compared to MI and Ad. Null. TREM-2 adenovirus transfection significantly activated the phosphorylation of AKT and mTOR compared to MI and Ad.Null transfection.
CONCLUSIONS Over-expression of TREM-2 by recombinant adenovirus transfection after MI alleviated the myocardial injury, decreased the infarcted size and inhibited the cardiomyocytes apoptosis. TREM-2 is a potential therapeutic target in improving heart function after MI.
GW31-e0018
Zhiteng Chen, Haifeng Zhang, Yangxin Chen, Jingfeng Wang
Sun Yat-sen Memorial Hospital
OBJECTIVES Cardiac fibrosis is a common pathophysiological process during different heart diseases, as well as an independent risk factor of heart failure prognosis. Fibroblast is the most important contributor to cardiac fibrosis. In various fibrotic diseases, glycolysis is proved to contribute to fibroblast activation. However, few researches focus on the glycolysis in cardiac fibroblast. As myocardial infarction is the main etiology of heart failure, it is of great significance to study the relationship among glycolysis, cardiac fibrosis and cardiac fibroblast activation.
METHODS In vivo, we delivered a glycolysis inhibitor, 2-deoxy-D-glucose (2-DG) or PBS by intraperitoneal injection after mice myocardial infarction and detected the cardiac function by ultrasonic cardiography, fibrosis area by Masson staining, cardiac fibroblast activation by tissue immunofluorescence, and glycolytic and cardiac fibroblast activation markers by western blot at the 28th day after myocardial infarction. In vitro, we activated human cardiac fibroblast (HCF) with transforming growth factor-β1 and detected the glycolytic change with Seahorse energy metabolism detector, lactate detection assay, glucose detection assay, and triphosadenine detection assay. We also detected the HCF activation markers with western blot and ELISA to confirm the change of glycolysis and the influence of 2-DG on HCF activation. At last, we also verified the relationship among cardiac fibrosis, cardiac fibroblast activation, and glycolysis in human fibrotic and non-fibrotic heart tissues.
RESULTS In vivo, we observe obvious cardiac fibrosis and fibroblast activation accompanied with enhanced glycolysis at the 28th day after myocardial infarction. We found an increase mortality in the 2-DG treated mice compared with the PBS treated mice. Then we delayed our initial administration time to the fourth day after myocardial infarction and lower the dosage of 2-DG to a half. Interestingly, the high mortality decreased, and the fibrosis and fibroblast activation in the heart were inhibited at the 28th day after myocardial infarction but with few benefits for cardiac function. In vitro, we uncovered that glycolysis was enhanced when cardiac fibroblast was activated. Glycolysis inhibition can alleviate the activation of cardiac fibroblast. Compared with non-fibrotic heart tissues, we uncovered that glycolysis increased together with fibrosis and fibroblast activation in human fibrotic heart tissues.
CONCLUSIONS Cardiac fibrosis is along with enhanced glycolysis. Glycolysis inhibition at the proper time and dosage can alleviate cardiac fibrosis progress and cardiac fibroblast activation after myocardial infarction.
GW31-e0019
Qiaozi Wang, Zheyong Huang, Junbo Ge
Zhongshan Hospital, Fudan
OBJECTIVES Heart diseases are the leading cause of death worldwide every year. Adult mammalian cardiomyocytes hold limited proliferative capacity so that regenerative therapies are needed. The direct reprogramming from fibroblasts into induced cardiomyocytes (iCMs) has shown much potential in the regenerative medicine, whereas its efficiency remains pretty low and latent safety issues may exist. Therefore, noninvasive intravenous delivery of key reprogramming-related factors to the injured heart might be the more ideal route. Fibroblasts within 3 days after acute myocardial infarction are the target cells of direct cardiac reprogramming. In this specific time window, neutrophils hold the strongest chemotaxis ability in the infarct area, as activated fibroblast surface glycoprotein TN-C secretion peaked. Recently nanoparticles have shown great benefit in gene delivery and in vivo targeting strategy. Our study aims to design a novel biomimetic nanoparticles-based layer-by-layer targeted system to deliver miRCombo to fibroblasts in vivo in order to realize reprogramming into iCMs.
METHODS Mesoporous silica nanoparticles (MSNs) were synthesized by chemical solution reaction. Neutrophil membrane protein (Neu) were obtained by ultracentrifugation. We assemble the FH-Neu-LiMSNs/miR by classical thin film hydration. Nanoparticles were injected by tain vein and the mice were killed after 2 weeks to evaluate the therapeutic effect.
RESULTS We successfully finished the package of nanoparticles, which were verified by transmission electron microscope. The core layer is mesoporous silica loaded with microRNAs, while biomimetic nanoliposomes of neutrophils with FH peptide coated outer layer. The nanoparticles were stable after freeze thawing and held relatively long half-life period in vivo. In vitro, the nanoparticles had the capacity of chemotaxis to chemotactic factor and could bind the injured fibroblasts by FH peptide. What’s more, these components did not influence the reprogramming effect of miRCombo in vivo. The nanoparticles could target the injured area after tail vein injection of MI model mice, revealed by in-vivo imaging system. After 2 weeks, histologic section fluorescence staining showed that regenerative iCMs could be observed in the infarcted area and had the similar structure with normal cardiomyocytes. Masson staining and echocardiography showed improved cardiac function and decreased fibrotic area.
CONCLUSIONS Our study showed that we successfully designed the novel nanoparticles to target the fibroblasts in the injured heart and could promoted cardiac regeneration and improve cardiac function after MI. Our system provides a new strategy for the regenerative medicine and holds great potential in clinical translation.
GW31-e0025
Zhen Wang, Di Ye, Jing Ye, Menglong Wang, Yao Xu, Jianfang Liu, Jishou Zhang, Mengmeng Zhao, Jun Wan
Renmin Hospital of Wuhan University
OBJECTIVES Sepsis-induced cardiac dysfunction is a common complication of sepsis and is associated with decreased survival of septic patients. Previous studies have demonstrated that interleukin (IL)-12p35 knockout regulates the progression of various cardiovascular diseases, such as acute myocardial infarction and hypertension. However, the effects of IL-12p35 on sepsis development remain unclear. Hence, this study aimed to determine the role of IL-12p35 in sepsis-induced cardiac dysfunction and explore its underlying mechanisms.
METHODS Lipopolysaccharide (LPS) was used to induce sepsis and myocardial injury, and the effect of LPS treatment on cardiac IL-12p35 expression was assessed. In addition, IL-12p35 knockout mice were used to determine the role of IL-12p35 in sepsis-induced cardiac dysfunction.
RESULTS First, we observed that LPS treatment significantly increased the cardiac expression level of IL-12p35. In addition, our findings demonstrated that IL-12p35 knockout mice exhibited higher serum and cardiac lactate dehydrogenase (LDH) levels, higher serum and cardiac creatine kinase-myocardial band (CK-MB) levels, and lower survival rates than LPS-treated mice. Moreover, IL-12p35 deletion further increased M1 macrophage differentiation and decreased M2 macrophage differentiation in LPS-treated mice. IL-12p35 deletion also downregulated the activity of AMP-activated protein kinase (AMPK) but increased the levels of phosphorylated p65 (p-p65) and phosphorylated NF-κB inhibitor alpha (p-IκBα).
CONCLUSIONS Knockout of IL-12p35 in mice aggravated LPS-induced cardiac injury and dysfunction by exacerbating the imbalance of M1 and M2 macrophages. These results suggest that IL-12p35 is an attractive target for treating sepsis-induced cardiac dysfunction.
GW31-e0028
Zijie Cheng, Lingmei Qian
Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University
OBJECTIVES Recent studies have revealed that proper exercise can reduce the risk of chronic disease and is beneficial to the body. Peptides have been shown to play an important role in various pathological processes, including cardiovascular diseases. However, little is known about the role of exercise-induced peptides in cardiovascular disease. We aimed to explore the function and mechanism of TAGLN peptide in ischemic injury and oxidative stress.
METHODS Cell viability, ROS, LDH, JC-1, TUNEL, Apoptosis were performed to evaluate the function of peptide TAGLN in vitro. Cardiac function, cardiac remodeling, HE staining, infarction size, Masson staining, cardiac markers were investigated to assess the function of peptide TAGLN in vivo. Pull-down, silver staining, Co-IP, Ubiquitination, Half-life and degradation were performed to analyze the mechanism of peptide TAGLN involved.
RESULTS Treatment with TAGLN peptide significantly improved cell viability, the mitochondrial membrane potential, and ROS levels and reduced LDH release, the apoptosis rate and caspase 3 activation in vitro. In vivo, TAGLN ameliorated MI and heart failure induced by I/R or DOX treatment. Pull-down assays showed that TAGLN can bind to PKG. The TAGLN-PKG complex inhibited PKG degradation through the UPS. We also identified cCbl as the E3 ligase of PKG and found that the interaction between these proteins was impaired by TAGLN treatment. In addition, we provided evidence that TAGLN mediated Lys48-linked polyubiquitination and subsequent proteasomal degradation.
CONCLUSIONS Our results reveal that a novel exercise-induced peptide, TAGLN, can inhibit PKG degradation by serving as a competitive binding peptide to attenuate the formation of the PKG-cCbl complex. Treatment with TAGLN may be a new therapeutic approach for MI.
GW31-e0054
Zheng Lian, Chen Hong
Peking University People’s Hospital
OBJECTIVES To explore the potential targets underlying the effect of rosuvastatin on heart failure (HF) by utilizing a network pharmacology approach and animal experiments to identify the results.
METHODS PharmMapper and other databases were mined for information relevant to the prediction of rosuvastatin targets and HF-related targets. Then, the rosuvastatin-HF target gene networks were created in Cytoscape software. Eventually, the targets and enriched pathways were examined by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Furthermore, we constructed an HF animal model and used rosuvastatin to treat them, identifying the changes in heart function and related protein expression.
RESULTS Thirty-five intersection targets indicated the therapeutic targets linked to HF. GO analysis showed that 481 biological processes, 4 cellular components and 23 molecular functions were identified. KEGG analysis showed 13 significant treatment pathways. In animal experiments, rosuvastatin significantly improved the cardiac function of post-myocardial infarction mice and prevented the development of HF after myocardial infarction by inhibiting IL-1β expression.
CONCLUSIONS The therapeutic mechanism of rosuvastatin against HF may be closely related to the inhibition of the expression of apoptosis-related proteins, inflammatory factors, and fibrosis-related genes. However, IL-1β is one of the most important target genes.
GW31-e0055
Dongjiu Li, Chengyu Mao, Changqian Wang
Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine
OBJECTIVES Post myocardial infarction (MI) fibrosis has been identified as an important factor in the progression of remodeling and heart failure. Previous studies have revealed that microRNA-21 (miR-21) played an important role in the pathogenesis of fibrosis. However, the exact role of miR-21 in post-MI fibrosis remains to be elucidated.
METHODS Wild type (WT) and miR-21 knockout (KO) mice were used and subjected to permanent left anterior descending coronary artery ligature.
RESULTS Wild type (WT) and miR-21 knockout (KO) mice were used and subjected to permanent left anterior descending coronary artery ligature. Compared with WT, miR-21 KO mice displayed smaller fibrotic area revealed by Masson’s trichrome staining as well as decreased expression of TGF-β, collagen Iα, α-SMA and Fibroblast activation protein (FAP), key markers of fibrotic process. In parallel, angiotensin II (Ang II) induced expression of α-SMA and FAP could be partially downregulated by miR-21 KO in mice primary cardiac fibroblasts (CFs). Mechanistically, we found that the expression of Sprouty1 (Spry1), a previously reported target of miR-21, was markedly elevated in miR-21 KO mice after MI, which could further inhibit the phosphorylation of extracellular-signal-regulated kinase (ERK1/2). In vitro study showed that Ang II promoted the phosphorylation of ERK1/2, activating TGF-β/Smad2/3 pathway. Phosphorylated Smad2/3 (p-Smad2/3) could further enhance the expression of α-SMA and FAP and may promote the maturation of miR-21, thus downregulating Spry1. In vitro knockdown of Spry1 by siRNA could rescue the effects of miR-21 inhibition on ERK/TGF-β/Smad2/3 signaling. Furthermore, in vivo inhibition of Spry1 could rescue the effects of miR-21 KO on cardiac fibrosis.
CONCLUSIONS These findings suggested that miR-21 promoted post MI fibrosis through targeting Spry1. Furthermore, miR-21 mediated a positive feedback on Ang II induced ERK/TGF-β/Smad pathway. Thus, targeting miR-21-Spry1 axis may be a promising therapeutic option for ameliorating cardiac fibrosis post MI.
GW31-e0069
Fang Wang1, Yimin Tu1, Yanxiang Gao1, Houzao Chen2, Jingang Zheng1
1Department of Cardiology, China-Japan Friendship Hospital, Beijing, China
2State Key Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China
OBJECTIVES Advancing age is the major risk factor of thoracic aortic aneurysm/dissection (TAAD). However, the causative link between age-related molecules and TAAD remains elusive. Here we first report a role of SIRT1, known as Class III histone deacetylase and the best studied member of Sirtuin family relevant to aging and longevity, in the prevention of TAAD in vivo.
METHODS Male SIRT1 smooth muscle specific transgenic (ST-Tg) mice, SIRT1 smooth muscle specific knockout (ST-KO) mice and their wild type (WT) littermates with C57BL/6J background were used to establish a TAAD model by oral administration of 3-aminopropionitrile fumarate (BAPN). The incidence and fatality rate of TAAD were analyzed between different groups. Western blotting and real-time PCR were utilized to examine matrix metallopeptidase 2 (MMP2) expression in aortas or cultured A7r5 cells. To clarify the epigenetic mechanism of MMP2 expression regulated by SIRT1 in vascular smooth muscle cells (VSMCs), chromatin immunoprecipitation (ChIP) assay was performed.
RESULTS BAPN treatment remarkably increased the incidence of TAAD in WT mice, which was attenuated in ST-Tg mice. Moreover, ST-KO promoted the fatality rate of TAAD induced by BAPN in mice. Mechanistically, SIRT1 overexpression reduced MMP2 level after BAPN treatment in both mouse aortas and cultured A7r5 cells. We further found that downregulation of BAPN-induced MMP2 expression by SIRT1 was mediated by deacetylation of histone H3 lysine 9 (H3K9) on Mmp2 promoter in vitro.
CONCLUSIONS We are the first to demonstrate that SIRT1 in VSMCs could be a novel therapeutic target for TAAD management.
GW31-e0100
Lishui Shen1, Xiaofeng Hu2, Yan Yao1
1Fuwai Hospital, Chinese Academy of Medical science, Peking Union Medical College
2Shanghai Chest Hospital, Shanghai Jiaotong University
OBJECTIVES Atrial fibrillation (AF) is one of the most common heart arrhythmic disorders all over the world. However, it is worth noting that the mechanism underlying AF is still dimness.
METHODS In this study, we implemented a series of bioinformatics methods to explore the mechanisms of lncRNAs underlying AF pathogenesis. The present study analyzed the public datasets (GSE2240 and GSE115574) to identify differentially expressed long non-coding RNAs (lncRNAs) and mRNAs in the progression of AF.
RESULTS Totally, 71 differentially expressed lncRNAs and 390 DEGs were identified in AF.Next, we performed bioinformatics analyses to explore the functions of lncRNAs in AF. Gene Ontology (GO) analysis indicated that differentially expressed lncRNAs were involved in regulating multiple key biological processes, such as cell cycle and signal transduction. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis demonstrated these lncRNAs were associated with the regulation of MAPK and Wnt signaling pathways. Eight lncRNAs (RP5-1154L15.2, RP11-339B21.15, RP11-448A19.1, RP11-676J12.4, LOC101930415, MALAT1, NEAT1, and PWAR6) were identified to be key lncRNAs and widely co-expressed with a series of differentially expressed genes (DEGs).
CONCLUSIONS Although further validation was still needed, our study may be helpful to elucidate the mechanisms of lncRNAs underlying AF pathogenesis and providing further insight into identifying novel biomarkers for AF.
GW31-e0101
Weijie Wang, Yao Zhang
The Second Affiliated Hospital of Harbin Medical University
OBJECTIVES With the development of research and application of anti-tumor drugs, the survival rate of cancer patients has increased year by year, but the cardiovascular disease problems caused by the use of anti-tumor drugs have become increasingly prominent. In recent years, the emerging discipline of oncology and cardiology has received more and more attention. Since the FDA approval in 2006, tyrosine kinase inhibitors (TKIs) have been clinically used to treat many types of cancer. Compared with previous drugs, they have a stronger anti-tumor effect. TKIs act as competitive inhibitors of tyrosine kinase binding to ATP, blocking the signal transduction of cell proliferation. Unlike trastuzumab and other drugs, some of the drugs in TKIs are multi-kinase target inhibitors, and other kinases that are not targeted to specific kinases may cause side effects, such as cardiovascular reactions. The study found that the direct effect of TKIs on cardiomyocytes may lead to heart failure, cardiomyopathy, conduction changes and prolonged QT intervals, leading to malignant arrhythmias until cardiac arrest. Based on the problem of anti-tumor drugs leading to cardiotoxicity, we explored the mechanism of targeting anti-tumor drugs tyrosine kinase inhibitors (TKIs)-lapatinib (cardatin toxicity), especially heart failure.
METHODS Through bioinformatics analysis, we integrated pharmacogenomics and GWAS catalog database to analyze genes related to lapatinib and heart failure. After that, we searched for related downstream targets on the string protein interaction website and predicted related signaling pathways.
RESULTS We integrated analysis of lapatinib and heart failure related genes through pharmacogenomics and GWAS catalog database, and obtained two SNP sites with higher scores, rs14213603 (score 2b, located in EGFR) and rs14145204 (score 3A, located ESR1). Through the String website to search for EGFR-related downstream pathway targets, we found that the proteins that interact most closely with EGFR, such as HRAS and EREG, are related to the ErBb signaling pathway. The specific kinase inhibition target of lapatinib is ErBb2. It is now found that ErBb2 is a molecule related to cardiac regeneration and has a protective effect on cardiomyocytes. Therefore, we believe that lapatinib inhibits EGFR and ErBb signaling pathways, causing off-target effects, inducing cardiomyocyte damage and leading to heart failure.
CONCLUSIONS (1) Lapatinib and human heart failure-related genes share a common SNP site. (2) Lapatinib inhibits EGFR and ErBb signaling pathways, causing off-target effects, inducing cardiomyocyte damage and leading to heart failure.
GW31-e0104
Meng Wang, Haifeng Zhang, Ganglan Fu, Minnan Gao, Lu Zhang, Huiqi Jiang, Jingfeng Wang, Yanqi Yang
Sun Yat-sen Memorial Hospital, Sun Yat-sen University
OBJECTIVES As one of the most common valvulopathies, calcific aortic valve stenosis (CAVS) has profound influence upon the aging population worldwide. Its progression involves complex immunological processes, molecular pathways and genetic factors. This research was conducted to comprehensively analyze the related immune cells and epigenic etiology in this biochemical process.
METHODS The data about CAVS were collected from GEO database and analyzed with R. The fastq files of RNA-sequencing data was processed with kallisto (0.44.0) in Linux and sleuth package (0.30.0) in R. The expression profiles from different platforms were combined to detect the immunological cellular components by using the online tools NetworkAnalyst and Cibersort. The weighted gene correlation analysis (WGCNA) was performed for each individual dataset. Subsequently a consensus network was constructed to identify the significantly altered gene modules. Then the results retrieved from NetworkAnalyst and WGCNA were compared to confirm the overlapped hub genes and enriched pathways. The immune cells infiltration pattern was related to the gene modules.
RESULTS Totally 66 samples in datasets GSE55492, GSE12644, GSE51472, GSE83453 were included and annotated as CAV group (n=33) and NAV group (n=33). In NetworkAnalyst 16,867 genes were matched for all data sets and 1494 different genes were identified. Cibersort showed macrophage M2 constituted 42.8% of all the immune cells in normal valve tissue and drop to 35.4% in calcified valve (P=0.0001). The hub genes FN1, VCAM1, IL7R, VAV1, BTK, PLAU, ARPC1B, CCL5, CDKN2A obtained from both WGCNA and NetworkAnalyst were related to CAVS positively. The hub genes CCL5, CDKN2A were positively correlated with regulatory T cells while negatively with M2 macrophage in calcified valves.
CONCLUSIONS The pathogenesis of CAVS involved extensive participation of immunological responses at cellular and molecular levels. The hub genes FN1, VCAM1, IL7R, VAV1, BTK, PLAU, ARPC1B, CCL5, CDKN2A and Toll-like receptor signaling, NF-kappa B signaling, ECM-receptor interaction, Focal adhesion, hematopoietic cell lineage pathways were closely linked to CAVS.
GW31-e0106
Wenjun Yan1, Chen Lin1, Yunlong Xia1, Xinliang Ma2, Ling Tao1
1Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, China
2Department of Medicine and Department of Emergency Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USA
OBJECTIVES Mesenchymal stromal cells (MSC)-based therapy is promising against ischemic heart failure (IHF). However, its efficacy is limited due to low cell retention and poor paracrine function. A transmembrane protein capable of enhancing cell–cell adhesion, N-cadherin garnered attention in the field of stem cell biology only recently. The current study investigates whether and how N-cadherin may regulate MSC retention and cardioprotective capability against IHF.
METHODS Adult mice-derived adipose tissue-derived MSC (ADSC) were transfected with adenovirus harboring N-cadherin (ADSC-Ncad), T-cadherin (ADSC-Tcad), or control adenovirus (ADSC-con). CM-DiI-labeled ADSC were intramyocardially injected into the infarct border zone at 3 sites immediately after myocardial infarction (MI) or myocardial ischemia/reperfusion (MI/R). ADSC retention/survival, cardiomyocyte apoptosis/proliferation, capillary density, cardiac fibrosis, and cardiac function were determined. Discovery-driven/cause-effect analysis was employed to determine the molecular mechanisms.
RESULTS Compared to ADSC-con, N-cadherin overexpression (but not T-cadherin) markedly increased engrafted ADSC survival/retention up to 7 days post-MI. Histological analysis revealed that ADSC-Ncad significantly preserved capillary density and increased cardiomyocyte proliferation, and moderately reduced cardiomyocyte apoptosis 3 days post-MI. More importantly, ADSC-Ncad (but not ADSC-Tcad) significantly increased LVEF and reduced fibrosis in both MI and MI/R mice. In vitro experiments demonstrated that N-cadherin overexpression promoted ADSC-cardiomyocyte adhesion and ADSC migration, enhancing their capability to increase angiogenesis and cardiomyocyte proliferation. MMP-10/13 and/or HGF upregulation is responsible for N-cadherin’s effect upon ADSC migration and paracrine angiogenesis. N-cadherin overexpression promotes cardiomyocyte proliferation by HGF release. Mechanistically, N-cadherin overexpression significantly increased N-cadherin/β-catenin complex formation and active β-catenin levels in the nucleus. β-Catenin knockdown abolished N-cadherin overexpression induced MMP-10, MMP-13, and HGF expression, and blocked the cellular actions and cardioprotective effects of ADSC overexpressing N-cadherin.
CONCLUSIONS We demonstrate for the first time that N-cadherin overexpression enhances MSC protective effects against IHF via β-catenin mediated MMP-10/MMP-13/HGF expression and production, promoting ADSC/cardiomyocyte adhesion and ADSC retention (Circulation Research. 2020;126:857–874, Cover paper, F1000Prime recommendation).
GW31-e0109
Xinpei Wang, Jiaxin Zhang, Changyang Xing, Xing Zhang, Yunchu Li, Feng Gao, Jia Li
Fourth Military Medical University
OBJECTIVES Dietary restriction has been well-described to improve health metrics, but whether it could benefit pathophysiological adaptation to extreme environment, e.g., microgravity, remains unknown. Here we investigated the effects of a daily rhythm of fasting and feeding without reducing caloric intake on cardiac function and metabolism against simulated microgravity.
METHODS Male rats under ad libitum feeding or time-restricted feeding (TRF; food access limited to 8 hours every day) were subjected to hindlimb unloading (HU) to simulate microgravity for 6 weeks. Left ventricular (LV) synchronicity and cardiac function were assessed by two-dimensional speckle tracking echocardiography. Glucose metabolites in the heart and blood were measured by liquid chromatography-tandem mass spectrometry.
RESULTS LV of HU rats displayed increased standard deviations of time to peak strain and maximum opposing wall delay, indicating a disturbed synchronicity compared with control rats. As a result, both cardiac systolic and diastolic function were declined in HU rats. In addition, HU inhibited glucose uptake and glucose oxidation by decreasing GLUT1 expression and inhibiting pyruvate dehydrogenase (PDH) activity in the heart, indicating an impaired cardiac metabolic flexibility. All these were largely rescued by TRF. Furthermore, TRF significantly improved contractile function of cardiomyocytes isolated from HU rats, although it showed no effects on HU-induced loss of cardiac mass. Interestingly, TRF raised liver-derived fibroblast growth factor 21 (FGF21) level and enhanced cardiac FGF21 signaling as manifested by upregulation of FGF receptor-1 expression and its downstream markers including phosphorylation of extracellular signal regulated kinase-1/2 and mRNA expression of PPARγ coactivator-1α in HU rats. In isolated cardiomyocytes, FGF21 treatment increased PDH activity, improved glucose utilization, and consequently enhanced cell contractile function. Finally, liver-specific knockdown of FGF21 expression by AAV8 carrying FGF21 shRNA abrogated the cardioprotective effects of TRF in HU rats.
CONCLUSIONS These data demonstrate that TRF ameliorates cardiac metabolic inflexibility and dysfunction induced by simulated microgravity through, at least partially, enhancing cardiac FGF21 signaling. Our data shed new lights on the cardiometabolic regulation of TRF and suggest TRF as a potential protective measurement for cardiovascular adaption to microgravity.
GW31-e0122
Bing Xiao, Xiuchun Yang
Department of Cardiology, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, China
OBJECTIVES To explore the effect of STIM1 inhibitor CM-4620 on endoplasmic reticulum (ER) calcium influx in macrophages, and its effect on oxidative stress induced by myocardial ischemia-reperfusion injury.
METHODS An in vivo model of myocardial ischemia-reperfusion (I/R) was established by using 30 C57BL/6J mice, which were divided into sham group, I/R group and CM-4620 group. Before surgery of 24 h, mice in CM-4620 group were treated 2 mM CM-4620 by tail vein injection and rats in sham and I/R groups were given same volume of saline. After surgery of 24 h, heart tissues were obtained. Masson staining was carried out to observe infiltration degree of myocardial tissues. DHE staining was used to detect ROS levels, and Tunel staining was used to detect cell apoptosis. An in vitro model was also established by using bone marrow-derived macrophages (BMDM), which were divided into control group, LPS group and LPS+CM-4620 group. Intercellular Ca2+ concentration was determined with Fura-2/AM fluorescent staining. The expression of STIM1, Orail, caspease-3, caspease-9, Bcl-2, Bax, IL-1beta, IL-6, TNF-alpha and IFN-gamma in both myocardial tissues and cells were detected by western blotting.
RESULTS In vivo model, the inflammatory infiltration degree, the ROS levels of myocardial tissues and the cell apoptosis in CM-4620 group were significantly reduced than those of I/R group (P<0.01). The expression of STIM1, orail, caspease-3, caspease-9, Bax, IL-1beta, IL-6, TNF-alpha and IFN-gamma were significantly lower in CM-4620 group than in I/R group (P<0.01), but the expression of Bcl-2 was significantly increased in CM-4620 group than in I/R group (P<0.01). In vitro model, CM-4620 significantly reduced the intercellular Ca2+ concentration and the ROS levels in LPS+CM-4620 group than in LPS group (P<0.01). The results of western blotting showed the decreased expression of STIM1, orail, caspease-3, caspease-9, Bax, IL-1beta, IL-6, TNF-alpha and IFN-gamma and the increased expression of Bal-2 in LPS+CM4620 group than in LPS group (P<0.01).
CONCLUSIONS CM-4620 can inhibit oxidative stress injury of myocardial ischemia-reperfusion by inhibiting calcium influx of macrophages endoplastic reticulum.
GW31-e0135
Yanbo Wang
The Second Hospital of Hebei Medical University
OBJECTIVES To study the influence of dl-3-N-butylphthalide (NBP) on infarction size in rats with acute myocardial infarction (AMI).
METHODS AMI model was established via ligation of left anterior descending artery. A total of 36 healthy male Sprague-Dawley rats (weight, 180±20 g), were randomly assigned to the sham-operation group (SO group, n=12), the model group (n=12) and the NBP group (n=12). The rats in the NBP group were treated with intraperitoneal injection administration of 60 mg/kg/body weight NBP once a day. The rats in the other groups were given distilled water of the same volume. The MI area in each group was detected via TTC staining. The concentrations of CK-MB and LDH were detected. The concentrations of TNF-α, IL-6, MDA and SOD were measured by ELISA.
RESULTS Compared with the SO group, the myocardial infarct sizes in the model group and the NBP group were significantly increased (P<0.001), and the infarct size in the NBP group was lower than that in the model group (280.6±5.82% vs. 37.74±10.18%, P<0.05). The levels of TNF-α in NBP group and model group were significantly increased compared with that in the SO group (<0.001), while the level of TNF-α in the NBP group was significantly lower than that in the model group (29.01±0.81 pg/mgprot vs. 37.72±0.96 pg/mgprot, P<0.05). The level of IL-6 in the model group and NBP group was significantly higher than that in the SO group, and it was lower than that in the model group (24.13±0.74 pg/mgprot vs. 27.53±1.03 pg/mgprot, P<0.05). The levels of MDA in the model group and the NBP group were significantly higher (<0.001), and the level of NBP group was lower than that in the model group (4.10±0.18 nmol/mgprot vs. 4.77±0.25 nmol/mgprot, P<0.05). The level of SOD in the model group and NBP group were lower (<0.001), and the SOD level in the NBP group was higher than that of the model group (53.49±3.89 U/mgprot vs. 38.06±5.28 U/mgprot, P<0.05).
CONCLUSIONS NBP could reduce the infarction size in SD rats with AMI.
GW31-e0139
Ningkun Zhang, Yu Chen, Li Zhao, Jiangchun He, Lihua Wang, Tianchang Li
Department of Cardiology, The Sixth Medical Center, Chinese PLA General Hospital
OBJECTIVES Investigate the effect of CTRP6 changes before and after coronary interventional surgery on patients with acute myocardial infarction (ami) and related factors of cardiovascular and metabolic disorders.
METHODS Selection of the sixth medical center of PLA general hospital during 2018–2019 in patients with acute myocardial infarction underwent emergency interventional therapy in 80 cases of normal control group selected volunteers check-up blood specimens of 20 cases, preoperative and postoperative patients with acute myocardial infarction (ami) venous blood was drawn, and the normal control group take a physical examination results of normal volunteers a venous blood and 4 c centrifugal 1200 g × 20 min, collect serum, using enzyme-linked immunosorbent assay (ELISA) method to detect CTRP6 level, collect basic data, detection of TC, TG, HDL-C, LDL-C, FBG and biochemical indexes such as, To analyze the correlation between changes before and after emergency intervention of CTRP6 and relevant clinical data.
RESULTS The age of patients with acute myocardial infarction (ami) was (52.7±12.6) years old, 57.8% of male patients and 42.2% of female patients, the smoking rate was 33.4%, the drinking rate was 35.7%, and the normal control group was (29.7±6.5) years old. In patients with acute myocardial infarction, preoperative CTRP6 (135.13±22.52) ng/mL was significantly lower than postoperative CTRP6 (227.44±37.28) ng/mL (P<0.01), and preoperative CTRP6 (135.13±22.52) ng/mL was significantly lower than the normal control group (219.38±31.68) ng/mL (P<0.01). After CTRP6 surgery (227.44±37.28) ng/mL showed no significant change compared with the normal control group (219.38±31.68) ng/mL (P>0.05). Correlation analysis showed that preoperative CTRP6 level was positively correlated with hdl-c; preoperative CTRP6 level was negatively correlated with BMI, TG, and FBG; preoperative CTRP6 level was not correlated with age, gender, smoking, drinking, TC, and ldl-c.
CONCLUSIONS CTRP6 in serum in patients with acute myocardial infarction was significantly reduced, terminate the risk factors in patients with acute myocardial infarction after CTRP6 rise and close to the control group, CTRP6 likely to be involved in the acute myocardial infarction (ami) and its related risk factors of metabolic development process. Studies show that members of the family of CTRP protein similar to adiponectin, members of the family of function CTRP in regulating the protection of cardiovascular disease, inflammation, and sugar play an important role in the regulation of lipid metabolism, the role of different members of the family of CTRP is different also, studies have shown that CTRP6 can significantly reduce the cardiac fibrosis after myocardial infarction, heart after infarction and TGF-β1 processing of fibroblasts, CTRP6 can inhibit myocardial fibroblasts differentiation and the production of extracellular matrix. In vitro knockout fibroblasts, CTRP6 enhanced the differentiation of TGF-TGF-β1 induced cardiac fibroblasts and the expression of various extracellular matrix proteins. In patients with cardiometabolic disorders, this study provides an experimental basis for whether CTRP6 can be used as an independent detection index to assist in the prediction of the occurrence of acute myocardial infarction.
GW31-e0146
Jieyun You1, Jian Wu2, Shijun Wang2, Fangjie Dai2, Yungzeng Zou2
1Department of Cardiology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China
2Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China
OBJECTIVES NLRP3 inflammation has emerged as an inflammatory cascade trigger contributing to cardiac remodeling induced by pressure overload. NLRP3 inflammasome related pyroptosis is a newly reported programmed cell death. As the pyroptosis executor, gasdermin D (GSDMD) forms pores on the lipid membrane, promotes the release of inflammatory cytokines and triggers inflammatory cascade. But its role remains to be elucidated in cardiac remodeling after pressure unloading.
METHODS By genetic interference with NLRP3 and GSDMD, we investigated the role of NLRP3/GSDMD-mediated pyroptosis on cardiac remodeling in mice subjected to transverse aortic constriction (TAC) for 4 weeks. Left ventricular structure and function were evaluated by echocardiographic, hemodynamic, and histological analyses. Signaling pathways related to pyroptosis, hypertrophy, fibrosis, angiogenesis, and apoptosis were analyzed by histological analysis, western blotting, ELISA, and real-time quantitative PCR.
RESULTS Our found that pressure overload upregulated the expression of NLRP3/GSDMD pathway, accompanied by the activation of inflammatory cytokines, and therefore deteriorated cardiac remodeling at least partially by activation of JNK1/2 and p38 signaling pathways. NLPR3 deficiency reversed the cleavage of GSDMD, leading to attenuation of cardiac remodeling and dysfunction induced by pressure overload. Further studies showed this process was independent of the Akt signaling pathway. On the other hand, overexpression of GSDMD abolished the cardioprotective effect of NLRP3 deficiency.
CONCLUSIONS Our data indicated that pressure overload upregulates NLRP3 to recruit inflammasome, therefore triggers pyroptosis and inflammation in a GSDMD-dependent manner, further activates JNK1/2 and p38 signaling pathways, and plays a critical role in cardiac remodeling induced by pressure overload. This study identified NLRP3/GSDMD as a promising therapeutic target for cardiac remodeling and heart failure.
GW31-e0147
Li Zhang, Lingmei Qian
The First Affiliated Hospital of Nanjing Medical University
OBJECTIVES The aim of this study is to elucidate the protective effect and mechanism of exercise-induced peptide EIP-22 in myocardial I/R injury, and to provide a new idea for the clinical treatment of ischemic heart disease.
METHODS We first determined the effect of EIP-22 on hypoxia/reoxygenation (H/R)- or H2O2-induced injury in the neonatal rat ventricular myocytes (NRVMs) via assessing cell viability and lactate dehydrogenase (LDH) level. In addition, TUNEL staining and western blot analysis of apoptosis related proteins were used to detect apoptosis level. Meanwhile, mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) accumulation was assessed by fluorescence microscope. Then we conducted rat I/R model and verified the effect of EIP-22 by measuring cardiac function, evaluating heart pathology and detecting serum LDH, CK-MB and cTnI concentrations. Finally, the main signaling pathway was analyzed by RNA-seq, and the activities of JAK2 and STAT3 were detected via western blot to illuminate molecular mechanism.
RESULTS In vitro, EIP-22 treatment significantly improved cells viability and MMP and attenuated the LDH release level, ROS accumulation, apotosis rate and the activation of apoptosis related proteins in NRVMs. In vivo, EIP-22 distinctly improved cardiac function, ameliorated myocardial infarction (MI) and decreased serum CK-MB and cTnI concentrations in rat I/R model. Mechanistically, JAK/STAT signaling pathway was focused by RNA-seq and we confirmed that treatment with EIP-22 elevated the expression of p-JAK2 and p-STAT3. Moreover, AG490, a selective inhibitor of JAK2/STAT3, eliminated the protective roles of EIP-22.
CONCLUSIONS The results uncovered that a novel exercise-induced peptide EIP-22 protected cardiomyocytes from myocardial I/R injury through regulating JAK2/STAT3 signalling pathway. This study suggested EIP-22 might be a new candidate molecule for the treatment of ischemic heart disease.
GW31-e0158
Yue Guo1,2, Xingfeng Xu1,2, Xinxue Liao1,2
1Department of Cardiology, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, PR China
2NHC Key Laboratory of Assisted Circulation (Sun Yat-sen University), Guangzhou, Guangdong 510080, PR China
OBJECTIVES The activation of NF-κB signaling pathway is regarded as the dominant process that correlates with the pathogenesis of diabetic cardiomyopathy (DCM). Recently, accumulating evidence shows that long noncoding RNAs (lncRNAs) play crucial roles in sustaining the NF-κB signaling pathway. However, the underlying mechanisms remain to be understood. In this study, we identified the upregulated expressed lncRNA NBR2 in adipocyte-derived exosomes (AdEXO) and investigated its regulatory role in diabetic myocardial fibrosis.
METHODS We examined the effect of exosomes from diabetic (db/db) mice-derived adipocytes on ANG-II-induced cardiac fibrosis and function in non-diabetic (C57BL/6J mice). In the in vitro study, HG (33 mmol/L) stimulated AdEXO were cultured with adult human cardiac fibroblasts (aHCFs). Differentially expressed lncRNAs in AdEXO were screened using lncRNA sequencing. The functional role of NBR2 was evaluated by both in vitro and in vivo experiments. Bioinformatics prediction, RNA fluorescence in situ hybridization (RNA-FISH) and immunoprecipitation assays were performed to identify the direct interactions between NBR2 and other associated targets, such as hnRNPK and SETDB1. Chromatin isolation by RNA purification assays were utilized to examine the interaction of NBB2 with IκBα promoter.
RESULTS Intramyocardial injection of diabetic adipocyte exosomes in the non-diabetic heart significantly exacerbated myocardial fibrosis, as evidenced by poorer cardiac function and enhancer collagen deposition. Whereas administration of a exosomes biogenesis inhibitor significantly mitigated cardiac fibrosis in diabetic mice. We identified that lncRNA-NBR2 is a common molecule significantly increased in diabetic Ad-EXO and HG-stimulated non-diabetic Ad-EXO. After four weeks of ANG II infusion, EXO-db/dbWT-injected mice displayed significant fibrosis in the heart; however, interestingly, mice receiving NBR2-deficient db/db-EXO showed a decrease in cardiac fibrosis. Similarly, AdEXO-NBR2 promoted aHCFs proliferation and transformation capabilities in vitro. Mechanistically, NBR2 was loaded to adipocyte-secreted exosomes by directly interacting with heterogeneous nuclear ribonucleoprotein K (hnRNPK). Subsequently, exosomal NBR2 was internalized by aHCFs and epigenetically downregulated IκBα expression by recruitment of hnRNPK/SETDB1 and increasing the H3K9 trimethylation level in the IκBα promoter, ultimately activating the NF-κB pathway.
CONCLUSIONS Our findings suggest that AdEXO-NBR2 provides a novel epigenetic mechanism involved in activation of NF-κB signaling pathway and may represent a new therapeutic target of DCM.
GW31-e0174
Sai Ye, Fangjun Guo, Renqiang Yang
Department of Cardiology, the Second Affiliated Hospital, Nanchang University, Nanchang 330006, China
OBJECTIVES To observe the influence of Ginkgolide B on myocardial fibrosis in rats with type 2 diabetes mellitus (T2DM) through the transforming growth factor beta-1 (TGF-β1)/Smad signaling pathway.
METHODS The rat model of type 2 diabetes mellitus was established by intraperitoneal injection of low dose streptozotocin (35 mg/kg) and high fat diet. The Sprague Dawley rats were randomly divided into control group (NC group), T2DM (DM group), and Ginkgolide B treatment group (GB group). GB group was established by intraperitoneal injection of a certain dose Ginkgolide B (10 mg/kg) for 12 weeks after successful model-making. The animals were euthanized, blood and myocardial tissues were collected from rats. Next, blood glucose level and body weight were detected. In addition, histopathological changes of the heart were observed with HE staining. The myocardial collagen volume fraction (CVF) was calculated by Masson staining. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Western blotting, which were applied to measure the gene and protein expression levels of important molecules in the proliferation and differentiation of myocardial fibroblasts including alpha-smooth muscle actin (α-SMA), and the relevant pathway TGF-β1, p-Smad2/3. Immunohistochemical method was performed to detect the depositions of collagen extracellular matrix proteins Col-I and Col-III.
RESULTS Compared with NC group, the body weight of the rat in DM group was decreased and the blood glucose was increased significantly (P<0.05). However, no statistically difference in blood glucose and body weight between the treatment groups and GB group was observed (P>0.05). As compared with NC group, CVF and the protein or mRNA levels of Col-III, TGF-β1, p-Smad2/3 and α-SMA were increased markedly in T2DM group (P<0.05). immunohistochemistry showed the expression levels of Col-I, Col-III collagen were increased (P<0.01). Moreover, CVF and the protein or mRNA levels of TGF-β1, Col-III, p-Smad2/3 and α-SMA in DM group were higher than treatment groups (P<0.05). immunohistochemistry showed the expression levels of Col-I, Col-III collagen in DM group were higher than treatment groups (P<0.05).
CONCLUSIONS Ginkgolide B has a protective effect on the myocardium of type 2 diabetes mellitus by inhibiting the TGF-β1/Smad2/3 signaling pathway, Ginkgolide B may be a novel medicine for treating diabetic cardiomyopathy.
GW31-e0208
Guangyin Shen1, Yisun Song2, Kyungsoo Kim2
1Department of Cardiology, Jilin Central Hospital, Jilin, China
2Division of Cardiology, Department of Internal Medicine, Hanyang University College of Medicine, Seoul, Republic of Korea
OBJECTIVES Previously, we reported that granulocyte-colony stimulating factor (G-CSF) was shown to reduce cardiomyocyte apoptosis in diabetic cardiomyopathy. However, the underlying mechanisms are not yet fully understood. Therefore, we investigated whether the mechanism of the anti-apoptotic effect of G-CSF was associated with autophagy in a rat model of diabetic cardiomyopathy.
METHODS Diabetic cardiomyopathy was induced in rats by a high-fat diet combined with low-dose streptozotocin and the rats were then treated with G-CSF for 5 days. Rat H9c2 cardiac cells were cultured under high glucose conditions as an in vitro model of diabetic cardiomyopathy. The extent of apoptosis and expression of proteins related to autophagy (Beclin-1, LC3-II/LC3-I ratio, and P62) were determined for both models.
RESULTS G-CSF significantly reduced cardiomyocyte apoptosis (25.12±4.24% vs. 34.51±3.93%, P<0.05) in the diabetic myocardium in vivo and led to up regulated expression of Beclin-1 (134.55±25.46% vs. 70.08±21.84%, P<0.05), and increased the LC3-II/LC3-I ratio (134.57±26.21 vs. 64.48±11.59, P<0.05), and down regulated expression of P62 (110.97±13.85% vs. 169.56±18.14%, P<0.05) compared with normal rats. Similarly, G-CSF suppressed apoptosis (17.70±5.41% vs. 29.50±3.93%, P<0.05) and up regulated expression of Beclin-1 (75.00±5.37% vs. 50.41±7.86%, P<0.05), increased the LC3-II/LC3-I ratio (119.36±14.37% vs. 75.07±5.41%, P<0.05), and down regulated expression of P62 (68.57±5.31% vs. 134.46±19.55%, P<0.05) in high glucose-induced H9c2 cardiac cells in vitro. These effects of G-CSF were abrogated by 3-methyladenine, an autophagy inhibitor.
CONCLUSIONS Our results suggest that G-CSF might reduce apoptosis through up regulation of autophagy.
GW31-e0221
ShiRan Yu1, Dong Xuefei2, Yang Min2, Cao Xinran2, Xiong Jie2, Dong Bo2
1Department of Cardiology, Center for Cardiovascular Translational Research, Beijing Key Laboratory of Early Prediction and Intervention of Acute Myocardial Infarction, Peking University People’s Hospital, Beijing 100044, China
2Department of Cardiology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China
OBJECTIVES In this study, we firstly aim to investigate whether excessive PRR in DCM could active YAP, further mediate downstream factors like CTGF and Smad, involving DCM pathological progression. Secondly, whether activated PRR-YAP pathway could be able to trigger cardiac redox reaction and cardiac fibrosis. Thirdly, whether an intervention targeted PRR-YAP pathway could alleviate these pathological injuries and delay disease’s progression.
METHODS Wistar rats and neonatal rat cardiac fibroblasts were respectively used in vivo and in vitro studies. In order to observe the effects of PRR mediated YAP pathway on the pathogenesis of DCM, animal experiments were divided into 3 parts, including the evaluation of PRR overexpression effects part, the assessment of the PRR RNAi silencing effects part and the evaluation of YAP RNAi silencing effects part. Recombinant-adenoviruses-carried-PRR-gene (Ad-PRR), recombinant-adenoviruses-carried-PRR-shRNA (Ad-PRR-shRNA) and lentivirus-carried-YAP-shRNA (LV-YAP-shRNA) were constructed and respectively injected these viruses in each group rats. Evaluating PRR and YAP expression in myocardium by immunohistochemical staining and western blotting to assess the efficacy of virus transfection, as well as the effects of PRR expression on YAP expression. Measured NADPH oxidase activity, MDA and SOD levels to explore the role of PRR-YAP pathway in redox reaction of DCM. Measured type I and III collagen expression by immunohistochemical staining to explore the role of PRR-YAP pathway in cardiac fibrosis of DCM. Measured CTGF and Smad3 expression by western blotting to explore underlying mechanism. In addition, in vitro experiments, using Ad-PRR and Ad-PRR-shRNA, meanwhile, introduce YAP specific inhibitor Verteporfin in cardiac fibroblasts along with PRR overexpression to further demonstrate the impact of PRR-YAP pathway on oxidative stress and myocardial fibrosis at cellular aspect.
RESULTS The results displayed that PRR activated YAP expressions and further exacerbated fibrosis and oxidative stress both in myocardium tissues and cardiac fibroblasts under high glucose condition. However, PRR RNAi silencing has opposite effects. Moreover, the administration of YAP inhibitor Verteporfin in vitro and YAP RNAi silencing in vivo could reverse PRR induced fibrosis and oxidative stress. And these effects may associate with the PRR-YAP pathway activate downstream factors expressions, including Smad3 and CTGF.
CONCLUSIONS We concluded that PRR-YAP pathway plays a key role in the oxidative stress and myocardial fibrosis in DCM and the treatment targeting PRR-YAP pathway may be a novel therapeutic way for DCM.
GW31-e0227
Shenjie Sun, Ping Zhang
Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University, Beijing
OBJECTIVES Owing to the complexity of the clinical phenotype and mechanism, the specific pathogenesis of atrial fibrillation (AF) is still not clear, especially at the genetic level. In search of etiologically guided solutions to improve diagnosis and treatment, in this study, we focused on the differentially expressed genes (DEGs) in AF using left atrial specimens and auricle tissue biopsy from patients to distinguish AF from sinus rhythm (SR) using bioinformatics approaches.
METHODS Microarray data from patient specimens were downloaded from two Gene Expression Omnibus (GEO) datasets (GSE79768 and GSE31821) to identify DEGs according to a log fold-change of >1 (upregulated genes) or >–1 (downregulated genes) and a P-value <0.05. We identified that the DEGs, the hub and key genes respectively from left atrial and auricle tissue. The DEGs were then used to construct a protein-protein interaction (PPI) network to further identify hub and key genes. Functional annotation of the differentially expressed genes was performed using Gene Ontology biological processes, cellular component, and pathway enrichment tools.
RESULTS Overall, we screened out 488 DEGs in GSE79768 from left atrial specimens, including 427 down-regulated genes and 61 up-regulated genes, and 414 DEGs in GSE31821 from the auricle tissue, including 392 down-regulated genes and 22 up-regulated genes. Function enrichment analysis using DAVID tools indicated that top five GO term plasma membrane, extracellular exosome, integral component of plasma membrane, extracellular region, and extracellular space in GSE79768 and top five GO term protein binding, cytosol, extracellular exosome, extracellular space, and cell surface in GSE31821. Further analysis indicated that the common DEGs were enriched in regulation of angiogenesis, cell surface, extracellular space, extracellular exosome, response to mechanical stimulus, and vasculature development. PPI network analysis mapped 758 nodes and 1516 edges in GSE79768, and 237 nodes and 934 edges in GSE31821. CXCR4, TLR4, and KIT were the top three hub genes in GSE79768 from left atrial specimens, whereas BMP4, CDH2, and BMP2 were the top three hub genes in GSE31821 from the auricle tissue. Finally, there is 16 hub genes is coexisting in GSE79768 and GSE31821. Among these, CXCR4 was identified as a key gene in the network (degree>10), with down-regulated expression in both the GSE79768 and GSE31821. PPI network analysis suggested that CXCR4 is the key gene associated with AF.
CONCLUSIONS The findings of our study suggest that the key gene CXCR4 may be associated with AF recurrence and maintenance and can be further explored as novel biomarkers in AF. Consequently, the top five miRNAs for CXCR4 may act as potential biomarkers or therapeutic targets for AF.
GW31-e0240
Junyi Yu1,2,3, Gengze Wu1,2, Xue Gong1,2, Hao Luo1,2, Zaicheng Xu1,2, Qiao Liao1,2, Zhi Chen1,2, Miao Tian1,2, Jining Yang4, Chen Gao3, Bing Zhang5, Christoph Rau3, Ye Zhang1,2, Yibin Wang3, Chunyu Zeng1,2
1Department of Cardiology, Daping Hospital, The Third Military Medical University, Chongqing, P.R. China
2Chongqing Institute of Cardiology, Chongqing, P.R. China
3Division of Molecular Medicine, Departments of Anesthesiology, Physiology and Medicine, David Geffen School of Medicine, University of California at Los Angeles (UCLA), Los Angeles, California, USA
4Research Center for Nutrition and Food Safety, Chongqing Key Laboratory of Nutrition and Food Safety, Institute of Military Preventive Medicine, Third Military Medical University, Chongqing, P.R. China
5Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai, China
OBJECTIVES Vascular remodeling is a major pathological feature under chronic hypertension. A subset of mammalian long non-coding RNAs (lncRNAs) are reported to encode functional peptides. Yet, in most cases, the coding products and the non-coding transcripts appear to function independently under unrelated mechanisms. Here we aim to illustrate the function and underlying mechanism of a novel peptide encoded by a lncRNA in vascular smooth muscle (VSMC) phenotype switching and vascular remodeling.
METHODS lncRNA-mRNA microarray was used to identify differentially expressed lncRNAs in aorta tissue from hypertensive vascular remodeling rats model. qRT-PCR and Western Blot were used to test RNA and protein expression respectively. CRISPR-Cas9 was used to construct protein ablation and gene knock-out rats. Carotid artery balloon injury was performed to induce vascular remodeling in vivo. siRNAs and lentivirus were used for gene knock-down, while plasmids and adenovirus were used for gene overexpression in vitro. CCK8 and Ki-67 staining were used to test the proliferation, while trans-well and wound scratching assays were used to test the migration of VSMC. RNA-seq was used to test the transcriptome alteration in VSMC. LC-MS was used to identify and co-IP was used to validate the protein interactions. ChIP-seq was used to test the protein-DNA interaction. ChIRP-seq was used to test the DNA-RNA interaction. RIP-PCR was used to validate the protein-RNA interaction. Dual luciferase assay was used to transcription regulation.
RESULTS We identified a conserved, VSMC enriched gene Phenotype-Switching-Regulator (PSR), which was previously annotated as a lncRNA. The PSR gene actually produced both a peptide (Arteridin) and a regulatory lncRNA (lncPSR). Arteridin and lncPSR were both necessary and sufficient to induce the transition of VSMCs from contractile to proliferative phenotype. Arteridin and lncPSR mediated VSMC gene regulation involved direct binding to a common transcription factor, YBX1. Arteridin modulates nucleic translocation of YBX1, and lncPSR directs chromatin targeting of YBX1 serving as a molecular scaffold. Intriguingly, the PSR gene transcription was also robustly induced by Arteridin, forming an auto-regulated feed-forward loop. Both Arteridin and lncPSR were significantly up-regulated in the remodeled arterial tissue from hypertension patients. Finally, both Arteridin protein ablation and PSR gene knockout attenuated neointima formation induced by carotid artery balloon injury in rats.
CONCLUSIONS This study has uncovered a previously uncharacterized genetic feed-forward regulatory circuit in vascular smooth muscle cell gene regulation that involves coordinated function of a regulatory lncRNA and its encoded peptide through a common transcription factor. Targeting Arteridin protein and lncPSR will provide ultimate therapeutic solution for vascular remodeling as well as other VSMC phenotype switching related diseases.
GW31-e0261
Jiahong Xue, Yuan Li, Fan Jiali, Wu Wenhuan
Department of Cardiovascular Medicine, the Second Affiliated Hospital, Xi’an Jiaotong University
OBJECTIVES Oxidative stress is an important factor that is related to endothelial dysfunction. ATP-binding cassette transporter G1 (ABCG1), a regulator of intracellular cholesterol efflux, has been found to prevent from endothelial activation by decreasing reactive oxygen species (ROS) production in vessel walls. However, the underlying mechanisms of ABCG1 inhibiting generation of ROS are still elusive.
METHODS Human Umbilical Artery endothelial cells (HUAECs) were transfected with specific ABCG1 siRNA or ABCG1 overexpression plasmid. Twenty four hours after transfection, cells were exposed to H2O2 to induce oxidative stress. Intracellular ROS production was measured using dihydroethidium, (DHE) fluorescent and malondialdehyde (MDA) was also tested. Prooxidant nicotinamide adenine dinucleotide phosphate (NADPH oxidase) activity was assessed by both lucigenin chemiluminescence assay and translocation of cytosolic p47phox to the membrane. Antioxidant signaling nuclear factor-erythroid 2 (Nrf2)/heme oxygenase 1 (HO-1) were observed by the migration of Nrf2 from cytoplasm to the nucleus via western blotting. Total intracellular cholesterol were measured with a microenzymatic fluorescence.
RESULTS The results showed that overexpression of ABCG1 by ABCG1 plasmid or T0901317 treatment inhibited ROS production and MDA content induced by H2O2 in HUAECs. Furthermore, ABCG1 upregulation blunted the activity of prooxidant NADPH oxidase and the expression of Nox4, one of NADPH oxidase subunit. Moreover, the increased migration of Nrf2 from cytoplasm to the nucleus and antioxidant HO-1 were all detected in HUAECs with upregulation of ABCG1. Conversely, ABCG1 downregulation by ABCG1 siRNA increased NADPH oxidase activity and Nox4 expression as well as abrogated the increase at Nrf2 nuclear protein levels. In addition, intracellular cholesterol load interfered with the balance between NADPH oxidase activity and HO-1 expression.
CONCLUSIONS It was suggested that ABCG1 attenuates oxidative stress induced by H2O2 in endothelial cells, which might involve in the balance between decreased NADPH oxidase activity and increased Nrf2/OH-1 antioxidant defense signaling via its regulation for intracellular cholesterol accumulation.
GW31-e0263
Yuanting Cui1,2, Hao Wu1, Qiang Li1, Zhiming Zhu1
1Department of Hypertension and Endocrinology, Center for Hypertension and Metabolic Diseases, Daping Hospital, Army Medical University, Chongqing Institute of Hypertension, Chongqing 400042, China
2Department of Cardiology, Southwest Hospital, Army Medical University, Chongqing 400038, China
OBJECTIVES Excessive salt intake is a major risk factor for hypertension and cardiovascular diseases. TRPM5 is a taste transient receptor potential cation channel selectively expressed in tongue epithelium. The aim of this study is to clarify the role of TRPM5 in modulation of salty-taste preference and salt intake.
METHODS The licking behavior of TRPM5 gene knockout (TRPM5KO) and wild-type (WT) mice were tested with a gustometer. The food intake and urinary sodium excretion were measured to estimate salt intake. The neuronal activity of the taste cortex was recorded through fiber fluorometry technique. The ambulatory blood pressure was measured by the telemetry system. The expression of TRPM5 was examined through immunoblotting analysis.
RESULTS TRPM5-induced high-salt taste perception was encoded by neurons in central bitter cortical field of the insula cortex. The expression of TRPM5 was lowered after a long-term high-salt dietary intervention. The mice fed on high-salt diet (HSD) exhibited an increased preference for high salt solutions than the mice on normal diet (ND). The mice on HSD displayed an increased preference for salt diet with 1% and 4% NaCl and a decreased preference for normal diet with 0.25% NaCl compared with the mice on ND. Accordingly, the salt intake and sodium excretion of HSD mice were higher than ND mice. After a long-term HSD administration, the blood pressure of TRPM5KO mice was higher than WT mice, and the secondary cardiac hypertrophy was more severe as well.
CONCLUSIONS These results indicate that the reduced expression of TRPM5 in tongue epithelium is a critical step of enhanced salty preference and hypertension in long-term high salt intake. Administration on TRPM5 may be a novel strategy for reducing high salt intake and blood pressure.
GW31-e0266
Xueqing Gan1,2, Shuang Li1, Dachun Yang1
1Department of Cardiology, The General Hospital of Western Theater Command
2Department of Cardiology, Xuyong People’s Hospital
OBJECTIVES Previous studies suggest that Rev-erbα has important roles in myocardial infarction, ischemia reperfusion, and heart failure, but the mechanism of Rev-erbα in neointimal hyperplasia after vascular endothelial injury remains unclear. We used Rev-erbα agonist SR9011 and antagonist SR8278 to investigate the role of Rev-erbα in neointimal hyperplasia after vascular endothelial injury.
METHODS We used Rev-erbα agonist SR9011 and antagonist SR8278 to investigate the role of Rev-erbα in neointimal hyperplasia after vascular endothelial injury. We also examined the mechanisms of action via in vitro experiments. We used angiotensin (Ang) II to induce cellular proliferation in vascular smooth muscle cells (VSMCs). We investigated whether Rev-erbα SR9011 or SR8278 affected cell proliferation.
RESULTS Activation of Rev-erbα with agonist SR9011 suppressed neointimal hyperplasia, whereas antagonist SR8278 treatment promoted neointimal hyperplasia after vascular endothelial injury. In vitro experiments, SR9011 induced significant inhibition of cell growth and reduced the expression of Nlrp3 induced by Ang II. SR8278 treatment promoted VSMC proliferation and the expression of Nlrp3. Nlrp3 knockdown reversed Ang II-induced VSMC proliferation, and this effect was not reduced by SR8278. After Nlrp3 knockdown SR8278 did not promote proliferation.
CONCLUSIONS The agonist SR9011 suppressed neointimal hyperplasia and inhibited VSMC proliferation, whereas the antagonist SR8278 promoted neointimal hyperplasia and VSMC proliferation. Rev-erbα has negative regulatory effects on neointimal hyperplasia after vascular endothelial injury and proliferation of VSMCs.
GW31-e0274
Qingyuan Gao1,2, Yangxin Chen1,2, Haifeng Zhang1,2, Zhiteng Chen1,2, Shaohua Wang1,2, Jingfeng Wang1,2
1Department of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, PRC
2Laboratory of Cardiac Electrophysiology and Arrhythmia in Guangdong Province, Guangdong 5101120, PRC
OBJECTIVES Cardiac fibroblast (CF) activation is pivotal in cardiac fibrosis. We predicted and constructed a CF-specific competing endogenous RNA (ceRNA) network. Potential functions related to fibrosis of hub genes in this ceRNA network were explored.
METHODS The Gene Expression Omnibus database was used to search for available datasets. Differentially expressed mRNAs (DE-mRNAs) and lncRNAs (DE-lncRNAs) were identified. microRNAs were predicted and validated. The ceRNA network was constructed and visualized by Cytoscape and ceRNA regulatory mechanisms were validated. Single Gene Set Enrichment Analysis (SGSEA) and comparative toxicogenomics database (CTD) were conducted to analyze the most associated pathways and diseases of DE-mRNAs in the ceRNA network. The functions of hub genes in the ceRNA network were validated by siRNA depletion.
RESULTS Two datasets, which respectively described differentially expressed genes in human CFs and failing ventricles, were used for analysis. Four hundred and twenty DE-mRNAs and 39 DE-lncRNAs, and 369 DE-mRNAs and 93 DE-lncRNAs were identified respectively in the two datasets. Thirteen DE-mRNAs with the same expression tendency were overlapped in two datasets. Expressions of 11 predicted microRNAs were validated differently. Only 2 of the DE-lncRNAs were paired to any one of the 11 microRNAs. Finally, 2 mRNAs (ADAM19 and TGFBI), 3 microRNAs (miR-9-5p, miR-124-3p, and miR-153-3p) and 2 lncRNAs (LINC00511 and SNHG15) constituted a ceRNA network. As shown by SGSEA and CTD, both ADAM19 and TGFBI were closely relative to the TGF-β1 pathway and cardiac fibrosis, respectively. Furthermore, siRNA depletion for 2 mRNAs or 2 lncRNAs can alleviate CF activation.
CONCLUSIONS ADAM19 and TGFBI were crucial genes in CF during CF activation and cardiac fibrosis and 2 lncRNAs and 3 microRNAs were involved in their regulations in a ceRNA crosstalk.
GW31-e0282
Chengwen Hang1, Yuanxiu Song1, Yanan Li2, Siyao Zhang2, Yun Chang2, Ming Cui1
1Peking University Third Hospital
2Anzhen Hospital
OBJECTIVES Myomesin-1 (encoded by MYOM1 gene) is a main structural linker of M-band in striated muscle fibrils and participates in sarcomere assembly. Studies have reported that MYOM1 can also be expressed in nucleus and probably plays a role in transcriptional regulation. Its alternative spliceosome EH-myomesin, a biomarker for dilated cardiomyopathy, has been found to be associated with myotonic dystrophy type 1 (DM1). Mutations of MYOM1 in familial hereditary hypertrophic cardiomyopathy suggested its potential role in striated muscle disease. Although the structural features of MYOM1 have been well explained in the past few years, its biological function are little known. Besides, the causal relationship and mechanisms underlying the MYOM1-related myopathies (especially in myocardium) still remain unknown. Therefore, it is of great significance to establish a reliable MYMO1 knockout model to study its biological function.
METHODS The CRISPR/Cas9 gene editing technology was used to establish a MYOM1 knockout human embryonic stem cell line (MYOM1-/- hESC), which was then induced into cardiomyocytes (MYOM1-/- hESC-CMs) in vitro. Immunofluorescence staining and flow cytometry were used to identify the morphological characteristics of hESC-CMs. Calcium imaging and video analysis were utilized to evaluate intracellular calcium and contractile function. RNAseq, RT-PCR and Western Blot were performed to examine the transcriptome differences between MYOM1-/- CMs and WT CMs and elucidate the potential molecular mechanisms of the knockout phenotype. Several drugs were used for early intervention to evaluate its effectiveness on improving the atrophic phenotype caused by myomesin-1 deficiency.
RESULTS MYOM1 was confirmed to be expressed both in nucleus and cytoplasm of cardiomyocytes at day 30 postinduction. Higher proportion of sarcomere disassembly accompanied with nonuniform sarcomere length was more obvious in KO CMs. The expression of genes anchored to MYOM1, including MYOM2, MYOM3, TTN, OBSCN and OBSL1 were significantly up-regulated in KO CMs. Moreover, the cell size of KO CMs showed significant reduction compared to WT CMs. Deficiency of myomesin-1 led to increased apoptosis but had no effect on proliferation of cardiomyocytes. Video analysis indicated decreased contraction duration and amplitude, with prolonged time to peak in KO CMs. Calcium imaging showed decreased amplitude, increased duration and higher abundance of Ca2+ in sarcoplasmic reticulum, which contributed to contractile dysfunction. Besides, total CaMKII protein was not different between two groups, whereas CaMKIId was markedly reduced in KO hESC-CMs. Transcriptome analysis of day 30 cardiomyocytes showed a great difference in calcium signaling pathway between two groups and a molecular signature of anti-muscle atrophy in KO CMs. Interestingly, KO CMs had higher expression of vascular and endothelial-related genes, while WT CMs had predominance of cardiac development-associated genes. More importantly, this knockout phenotype can be partially attenuated by verapamil.
CONCLUSIONS MYOM1 plays an important role in sarcomere assembly, contractility regulation and cardiomyocytes development. MYOM1-/- hESC-CMs can recapitulate cardiac atrophy phenotype in vitro due to calcium disorders. Based on this model, the etiology, pathogenesis, and potential treatments of cardiac atrophy caused by myomesin-1 deficiency can be studied.
GW31-e0284
Yuping Fu1, Tiannan Jiang2, Qiangsun Zheng1
1The Second Affiliate Hospital of Xi’an Jiaotong University
2Beijing Friendship Hospital, Capital Medical University
OBJECTIVES Atrial fibrillation (AF) is a highly prevalent arrhythmia in clinical practice, and it is well accepted that aerobic exercise improves AF symptoms and -related quality of life. Necroptosis, a novel programmed cell death, plays a critical role in the development of fibrosis, yet it is largely unknown whether necroptosis contributes to AF and its role in exercise-conferred benefits on AF.
METHODS Mice were administrated with calcium chloride and acetylcholine (CaCl2-Ach) for 3 weeks to establish a model of AF. CaCl2-Ach-treated mice were either sedentary or subjected to 3-week swim training to investigate the effect of aerobic exercise on AF. AF susceptibility, heart structure and function and atrial fibrosis were assessed by electrophysiological examinations, echocardiography, Masson’s trichrome staining.
RESULTS Three-week CaCl2-Ach administration enhanced AF susceptibility as evidenced by increased AF frequency and duration time after burst pacing. Echocardiographic and histologic results also showed left atrial enlargement and fibrosis in AF mice, demonstrating apparent atrial structrual remodeling. Moreover, we found key mediators of necroptotic signaling (RIP1, RIP3, MLKL, CaMKII) were markedly activated in the atria of AF mice, while inhibiting necroptosis with necrostatin-1 partly attenuated CaCl2-Ach-induced fibrosis and AF susceptibility, indicating necroptosis plays a critical role in AF pathogenesis. Finally, we found 3-week swim training inhibited necroptotic signaling, thereby decreased CaCl2-Ach-induced AF susceptibility and atrial structural remodeling.
CONCLUSIONS Our findings identify necroptosis as a novel mechanism in AF pathogenesis and highlight that aerobic exercise may confer benefits on AF via inhibiting cardiac necroptosis.
GW31-e0291
Kun Zhao, Peng Li, Yong Li
The First Affiliated Hospital of Nanjing Medical University
OBJECTIVES A high salt diet (HSD) is a key risk factor for hypertension, and nitric oxide attenuates cardiac damage. The present study aimed to explore whether nitric oxide could alleviate high salt-induced apoptosis and autophagy of cardiomyocytes in rats.
METHODS Rats received 8% HSD in vivo. H9C2 cells or primary neonatal rat cardiomyocytes (NRCM) were treated with sodium chloride (NaCl) in vitro.
RESULTS Middle and high doses (50 mM and 100 mM) of NaCl increased the level of cleaved-caspase 3/caspase 3, cleaved-caspase 8/caspase 8, Bax/Bcl2, and LC3 in H9C2 cells. By contrast, 25 mM NaCl and 100 mM mannitol exerted no effect on the levels of cleaved-caspase 3/caspase 3, cleaved-caspase 8/caspase 8, and Bax/Bcl2, or LC3 compared with PBS in H9C2 cells. The endothelial nitric oxide synthase (eNOS) level was increased in the heart of HSD rats and H9C2 cells treated with 100 mM NaCl. Nitric oxide (NO) donor sodium nitroprusside (SNP) reduced the increased levels of cleaved-caspase 3/caspase 3, cleaved-caspase 8/caspase 8, Bax/Bcl2 induced by NaCl (100 mM) in H9C2 cells and NRCM. The levels of cleaved-caspase 3/caspase 3, Bax/Bcl2, and LC3 were increased in the heart of HSD rats. SNP treatment attenuated the increases of cleaved-caspase 3/caspase 3, Bax/Bcl2, and LC3 in the heart of HSD rats.
CONCLUSIONS Overall, our study demonstrated that NaCl increased apoptosis and autophagy of cardiomyocytes, and eNOS/NO alleviated the high salt-induced apoptosis and autophagy of cardiomyocytes.
GW31-e0292
Kun Zhao, Chuanxi Yang, Peng Li, Wei Sun, Xiangqing Kong
The First Affiliated Hospital of Nanjing Medical University
OBJECTIVES Cardiac fibrosis and hypertrophy, as the major hallmarks of cardiac remodeling involved in the pathophysiological process of hypertensive heart diseases, can result in disturbed function and structure of the myocardium, accompanied by the increased deposition of myocardial collagen. Recent studies have demonstrated that alamandine, the heptapeptide in the Ang-(1-7)//MrgD axis, could alleviate AngII-induced cardiac hypertrophy through binding to its natural receptor Mas-related G protein-coupled receptor member D (MrgD). However, no data to date demonstrates the specific physiological and pathophysiological function of MrgD in the therapeutic effect of alamandine towards AngII induction. Thus, we aimed to investigate the role of MrgD in regulating AngII-induced cardiac hypertrophy in vivo and in vitro.
METHODS In our study, we used AngII to mimic the animal or cell culture models of cardiac hypertrophy and fibrosis. After alamandine treatment, the nucleoproteins were collected to determine the role of alamandine in AngII-induced nuclear import. Pretreatment or intra-myocardial injection of recombinant adenovirus-MrgD (AD-MrgD) or adenovirus-shRNA-MrgD (shRNA-MrgD) were further used to verify the pathophysiological function of MrgD in vivo and in vitro. Then, LIPUS irradiation (0.5 MHz, 77.20 mW/cm2) was applied for 20 minutes every other day in mice received chronic AngII infusion in vivo. Following that, the levels of cardiac hypertrophy and fibrosis were evaluated by echocardiographic, histopathological, and molecular biological methods.
RESULTS Our results showed the protein and mRNA expression levels, as well as the fluorescence intensities of MrgD were increased after alamandine and/or AngII treatment, while there was no significant difference in those between these two groups. Further, more nuclear import of MrgD was found in cells receiving AngII stimulation by analyzing the results of nucleoproteins compared to those after alamandine treatment alone in vitro. Interestingly, alamandine treatment could reduce AngII-induced MrgD nuclear import by inactivating the phosphorylation of PKA and alleviating oxidative stress, which may account for the beneficial effects of alamandine towards AngII. Then, adenovirus silencing MrgD pre-treatment which reduced the total protein and nuclear protein expressions of MrgD in vitro could mimic the protective role of alamandine, while adenovirus-mediated overexpression of MrgD aggravated the pathological effects induced by AngII in vivo and in vitro. More importantly, we further found that low-intensity pulsed ultrasound (LIPUS) could ameliorate AngII-induced cardiac fibrosis via decreasing MrgD expression in vivo and in vitro.
CONCLUSIONS Taken together, our current study unveiled the promising cardioprotective effect of silencing MrgD expression on alleviating AngII-induced cardiac hypertrophy and fibrosis by reducing its nuclear import, paving the way to develop novel therapeutic apparatus, LIPUS, in the clinical practice of cardiac remodeling in the future.
GW31-e0293
Kun Zhao, Chuanxi Yang, Peipei Huang
The First Affiliated Hospital of Nanjing Medical University
OBJECTIVES Pellino1 has been shown to regulate proinflammatory genes by activating the nuclear factor kappa B (NF-κB) and Toll-like receptor (TLR) signaling pathways, which are important in the pathological development of lipopolysaccharide (LPS)-induced myocarditis. However, it is still unknown whether silencing Pellino1 (si-Pellino1) has a therapeutic effect on this disease. Here, we showed that silencing Pellino1 can be a potential protective strategy for abnormal myocardial energy metabolism in LPS-induced myocarditis.
METHODS We used liquid chromatography electrospray-ionization tandem mass spectrometry (LC-MS/MS) to analyze samples from si-Pellino1 neonatal rat cardiac myocytes (NRCMs) treated with LPS or left untreated. After normalization of the data, metabolite interaction analysis of matched KEGG pathway associations following si-Pellino1 treatment was applied, accompanied by interaction analysis of gene and metabolite associations after this treatment. Moreover, we used western blot (WB) and polymerase chain reaction (PCR) analyses to determine the expression of genes involved in regulating cardiac energy and energy metabolism in different groups.
RESULTS LC-MS-based metabolic profiling analysis demonstrated that si-Pellino1 treatment could alleviate or even reverse LPS-induced cellular damage by altering cardiac energy metabolism accompanied by changes in key genes (Cs, Cpt2, and Acadm) and metabolites (3-oxoocotanoyl-CoA, hydroxypyruvic acid, lauroyl-CoA, and NADPH) in NRCMs.
CONCLUSIONS Overall, our study unveiled the promising cardioprotective effect of silencing Pellino1 in LPS-induced myocarditis through fuel and energy metabolic regulation, which can also serve as biomarkers for this disease.
GW31-e0358
Cheng Li, Shudong Wang, Jian Sun, Quan Liu, Yonggang Wang
Department of Cardiovascular Center, The First Hospital of Jilin University, Changchun, Jilin 130021, China
OBJECTIVES Diabetic cardiomyopathy (DCM) – pathophysiological heart failure that occurs in absence of coronary artery disease, hypertension, and/or valvular heart disease, is a common diabetic complication. Elabela, a new peptide that acts via APJ, has similar functions as Apelin, providing beneficial effects on body fluid homeostasis, cardiovascular health, renal insufficiency, as well as potentially beneficial effects on metabolism and diabetes. The present study sought to reveal the underlying mechanisms of Elabela mediated cardioprotection.
METHODS Six weeks old, male C57BL/6J mice were randomized into 4 groups: (1) Control (Ctrl) group; (2) Ctrl+Elabela group; (3) Diabetes mellitus (DM) group; (4) DM+Elabela group. Type I diabetes model mice was established by intraperitoneally injecting with streptozotocin (150 mg/kg). Elabela (4.5 mg/kg) was administered subcutaneously twice a day for 3 months. Body weight, plasma glucose levels, cardiac function and structure were measured at the end of the experiment. Masson trichrome and TUNEL staining were used to assess myocardial fibrosis and apoptosis, respectively. The expressions of fibrosis, oxidative stress, inflammation, and apoptosis molecules were measured by quantitative PCR and/or Western blot. Furthermore, the expressions of SIRT3, its interaction protein Foxo3a, and its downstream anti-oxidative proteins SOD-2 and MnSOD were also measured.
RESULTS Neither body weight nor plasma glucose levels was affected by Elabela. Cardiac fibrosis, oxidative stress, inflammation, and apoptosis were increased in DM group compared to Ctrl group, all of which were attenuated by Elabela treatment. In addition, the expressions of the SIRT3 and its downstream anti-oxidative proteins SOD-2 and MnSOD were lower than that in Ctrl group, which can be significantly improved by Elabela treatment. Especially, we observed that diabetes significantly increased the level of acetylated Foxo3a while diabetes treated with Elabela decreased acetylated expression.
CONCLUSIONS In this study, Elabela treatment was found to have profound protective effects against diabetes-induced cardiac fibrosis, oxidative stress, inflammation, and apoptosis; these protective effects may depend heavily upon SIRT3-mediated Foxo3a deacetylation. Our findings provide evidence that Elabela has cardioprotective effects for the first time in the diabetic model.
GW31-e0364
Yuanxiu Song, Chengwen Hang, Ming Cui
Department of Cardiology, Peking University Third Hospital
OBJECTIVES Cardiac remodeling is the initial factor of heart failure caused by various causes. It is closely related to the abnormal energy metabolism of myocardial cells caused by abnormal mitochondrial dynamics. MTFR1L is a member of the mitochondrial dynamic network, which can promote mitochondrial division. There is no research on the specific physiology of MTFR1L and its pathology related to cardiac remodeling. This study used the combination of iPSC myocardial differentiation and CRISPR/Cas9 gene editing technology to establish a human homozygous knockout cardiomyocyte model of MTFR1L, and explored the relationship between MTFR1L and cardiac remodeling and its molecular mechanism.
METHODS The author combined CRISPR/Cas9 genome editing technology with myocardial differentiation technique of human induced pluripotent stem cells (hiPSCs) to establish the MTFR1L-/- cell line and differentiated into cardiomyocytes. RNA-seq was used to analyze differential genes in wild-type (WT) and MTFR1L-/- (KO) hiPSC-CMs, and key molecules and signaling pathways that MTFR1L regulated cardiac remodeling were screened out. Immunofluorescence staining, electron microscopy and flow cytometry were used to observe the effect of MTFR1L knockout on myocardial cell morphology. Mitotracker and Mitosox fluorescent probes were used to observe the effect of MTFR1L on myocardial mitochondrial number and ROS level. Flou-4/AM probe was used to explore the role of MTFR1L on myocardial cell calcium signal and contractility. Annexin V/APC fluorescence apoptosis detection was used to compare the difference in apoptosis between WT and KO hiPSC-CMs. Western blot and qPCR were used to verify the expression differences of heart failure, fibrosis, ion channels, apoptosis and autophagy-related pathway genes in cardiomyocytes.
RESULTS Compared with WT hiPSC-CMs, the cell volume became smaller, the number of mitochondria decreased, and the length increased (P<0.05), indicating that MTFR1L deficiency doesn’t affect the differentiation of myocardium, but it can make mitochondrial and cell morphology changes. The knockout of MTFR1L caused the increase of mitochondrial ROS level and the decrease of mitochondrial DNA content (P<0.05) in myocardial cells, resulting in mitochondrial damage. QPCR results showed KO hiPSC-CMs ion channel related genes (SCN5A, KCNH2, KCNQ1) expression was significantly reduced, while fibrosis related genes (Col1a, ATP2a) and other cardiac remodeling-related genes increased significantly (P<0.05); MTFR1L deficiency caused myocardial calcium activity disturbance (P<0.05). RNA-seq results showed that the expression of apoptosis-related pathway genes (caspase8, capase10) and autophagy pathway-related genes (LAMP1, LAMP2, MAPK10) in KO hiPSC-CMs increased significantly. The results of apoptosis detection showed the loss of MTFR1L promotes the level of cardiomyocyte apoptosis.
CONCLUSIONS Mitochondrial division-related gene MTFR1L plays an important role in maintaining mitochondrial morphology and function intact, participating in the maintenance of cardiomyocyte function, and inhibiting cardiac remodeling. This effect is achieved by maintaining mitochondrial division-fusion balance in cardiomyocytes, preventing apoptosis and excessive activation of autophagy. This research helps elucidate the molecular mechanism of heart remodeling and provides new ideas for the prevention and treatment of myocardial mitochondrial energy metabolic diseases.
GW31-e0390
Xiaoping Wang, Yong Wang
Beijing University of Chinese Medicine
OBJECTIVES Doxorubicin (DOX) is an effective first-line chemotherapeutic agent that is widely used in the treatment of various cancers. However, accumulation of DOX can cause side effects with the cardiotoxicity as the most severe one. Oxidative stress and cardiomyocyte apoptosis play a key role in DOX-induced cardiotoxicity (DIC). DOX could induce oxidative stress, which leads to opening of the mitochondrial permeability transition pore (mPTP) and apoptosis in cardiomyocytes. Opening of mPTP is regulated by cyclophilin D (CypD) and glycogen synthase kinase 3β (GSK-3β). Phosphorylation of GSK-3β by AKT could promote its interaction with adenine nucleotide translocator (ANT), inhibit opening of mPTP and prevent apoptosis. Previous studies have shown that cryptotanshinone (Cts) has potential cardioprotective effects, but its role in DIC remains unknown. Our research aimed to explore whether Cts could ameliorate DOX-induced oxidative and apoptosis by targeting Akt-GSK-3β-mPTP pathway.
METHODS A DOX-stimulated H9C2 cell model was established in this study. The effects of Cts on cell viability, reactive oxygen species (ROS) level, superoxide ion accumulation, apoptosis and mitochondrial membrane potential (MMP) were evaluated. Expressions of proteins in Akt-GSK-3β pathway were detected by western blot. Akt inhibitor was applied to treat cells to investigate the effects of Cts on Akt-GSK-3β pathway and mPTP. The effects of Cts on the binding of p-GSK-3β to ANT and the formation of ANT-CypD complex were explored by immunoprecipitation assay.
RESULTS The results showed that Cts could increase cell viability, reduce ROS level, inhibit apoptosis and protect mitochondrial membrane intergrerity. Cts treatment increased phosphorylated levels of Akt and GSK-3β. After cells were co-treated with Akt inhibitor, the effects of Cts on phosphorylation of GSK-3β, ROS, apoptosis and MMP were also abolished. Immunoprecipitation assay showed that Cts significantly increased the GSK-3β-ANT interaction and attenuated DOX-induced formation of ANT-Cyp-D complex, thereby inhibiting opening of mPTP.
CONCLUSIONS In conclusion, Cts is a promising drug for preventing DIC and it ameliorates oxidative stress and apoptosis by acting on Akt-GSK-3β-mPTP pathway.
GW31-e0394
Haifeng Zhang1,2, Shaohua Wang1,2, Qingyuan Gao1,2, Zhiteng Chen1,2, Guanghao Gao1,2, Yangxin Chen1,2, Jingfeng Wang1,2
1Department of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, PRC
2Laboratory of Cardiac Electrophysiology and Arrhythmia in Guangdong Province, Guangdong 5101120, PRC
OBJECTIVES To explore the effects of vitamin (Vit)-C in combination with endothelin (ET)-1 on embryonic stem cell (ESC) differentiation into cardiomyocytes and underline mechanism.
METHODS Different Vit-C concentrations and intervention time were tested to optimize Vit-C treatment. After that, Vit-C (100 μmol/L) alone or together with ET-1 (100 nmol/L) were administered to ESC at day 4–9 after differentiation. The beating of embryoid bodies (EBs) was calculated and compared between groups. Expressions of cardiac marker, cTnT, cardiac promotor, Nkx 2.5, GATA-4, and Shox2 were determined. Lentivirus mediated RNA interference was used to knockdown expressions of Nkx 2.5, GATA-4, and Shox2 and EBs beating under Vit-C and ET-1 treatment were observed.
RESULTS Optimal Vit-C treating concentration was 100 μmol/L and day 4–9 after differentiation was the best time to intervene, which increased EBs beating by 11.19% and increased expression of cTnT, Nkx 2.5, GATA-4, and Shox2 expressions at the same time. In combination with ET-1 further increased EBs beating by 11.07% and expression profiles of the above proteins. Knocking down of Shox2 reversed the effects of Vit-C and ET-1 at the greatest extent, reduced EBs beating by 12.94%.
CONCLUSIONS Vit-C plus ET-1 produce more purified cardiomyocytes from ESC differentiation. The up-regulation of Shox2 is the most important underline mechanism.
GW31-e0399
Yujie Xing, Jing Xu, Shuo Pan, Junkun Wang
Department of Cardiology, Shannxi Provincial People’s Hospital, Xi’an, Shaanxi
OBJECTIVES To explore the effects of selenium deficiency on cardiac function and myocardial tissue in SD rats.
METHODS Sixty SD rats were randomly divided into control group, selenium deficiency group and selenium supplement group, with twenty rats in each group. Rats in the control group were fed with standard diet, rats in the selenium deficiency group were fed with selenium deficiency diet and rats in the selenium supplement group were fed with selenium deficiency diet for fourteen weeks and then giving sodium selenite for three weeks. Seventeen weeks later, the levels of blood selenium and BNP were detected. HE staining was used to detect the morphological change of myocardial tissue. The electron microscope was used to detect the myocardial ultrastructure. The cardiac function in rats was detected by the echocardiography.
RESULTS The levels of blood selenium in selenium deficiency group decreased significantly compared with the control group, but after selenium supplement they increased significantly. Compared with the control group, the BNP level of rats in the selenium deficiency group was higher, but the BNP level was significantly decreased after selenium supplement. In the control group the structure of myocardial tissue was normal, but the structure of myocardial tissue in the selenium deficiency group was disordered. There was only mild abnormality in the structure of myocardial tissue in the selenium supplement group. Compared with the control group, the cardiac function of rats in the selenium deficiency group was significantly reduced, but the cardiac function was significantly increased after selenium supplement.
CONCLUSIONS Selenium deficiency can reduce the cardiac function and destroy the normal structure of myocardial tissue. Meanwhile, selenium supplement can improve cardiac function and improve the destruction of myocardial tissue structure in rats.
GW31-e0403
Qingyuan Gao1,2, Haifeng Zhang1,2, Zhiteng Chen1,2, Shaohua Wang1,2, Yangxin Chen1,2, Jingfeng Wang1,2
1Department of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, PRC
2Laboratory of Cardiac Electrophysiology and Arrhythmia in Guangdong Province, Guangdong 5101120, PRC
OBJECTIVES The downstream regulators of TGF-β1 are attractive in the studies of cardiac fibrosis and cardiac fibroblast (CF) activation. Imbalanced mitochondrial dynamics were investigated in aged-related diseases and other organ fibrosis. The potential role of Dynamin related protein-1 (Drp1)-induced excessive mitochondrial fission in TGFβ1-stimulated CF activation was explored.
METHODS Mouse CF were isolated from 1–3 days old C57BL/6 neonatal mice that received TGF-β1 (5 ng/mL) or saline. Mitochondrial morphology, expression of the regulatory molecules in mitochondrial fission and fusion, and Drp1-Tom20 interactions were measured. Also, mitochondrial membrane potential and mitochondrial ROS production were measured to reflect mitochondrial damage. The Drp1 inhibitor mitochondrial division inhibitor 1 (Mdivi-1) and mitochondrial fusion promotor M1 (Promotor M1) were added in the TGFβ1-induced group to assess the changes of pro-fibrotic markers, collagen deposition, proliferation, and migration in activated CF. The cell metabolism throughout mitochondrial respiratory and glycolysis were measured by Seahorse XFe96 Analyzer.
RESULTS More mitochondrial fission and less mitochondrial fusion events happened in TGFβ1-induced CF. The expressions of pro-fibrotic markers, collagen secretion and deposition, and CF proliferation and migration can be mitigated throughout inhibiting Drp1-induced mitochondrial fission by Mdivi-1 or promoting mitochondrial fusion by Promotor M1. Besides, this rescuing experiment was accompanied by alleviation in mitochondrial damage. On the other hand, through detecting the level of cell metabolism, Drp1-induced mitochondrial fission in TGFβ1-stimulated CF can lead to impaired mitochondrial respiratory and enhanced glycolysis. The latter can cause CF activation and cardiac fibrosis. 2-Deoxy-D-glucose, a glycolysis inhibitor, can further rescue the TGFβ1-induced CF activation based on Mdivi-1 or Promotor M1-treated activated CF.
CONCLUSIONS Excessive Drp1-induced mitochondrial fission and impaired mitochondrial fusion can cause CF activation throughout enhancing glycolysis in TGFβ1-induced CF.
GW31-e0429
Ma Lin, Wang Yong
Beijing University of Chinese Medicine
OBJECTIVES Neocryptotanshinone (NCTS) is a natural molecule identified from traditional Chinese herb Salvia miltiorrhiza Bunge, which is widely used in the treatment of cardiovascular diseases. RXRα is an attractive target due to its impressive effect on the heart energy metabolism. This study is to investigate whether the protective effect of NCTS on the heart failure (HF) was exerted via regulating RXRα-mediated energy metabolism pathway.
METHODS Microscale Thermophoresis (MST) were used to verify the binding force of NCTS to the target RXRα. The model of HF post-acute myocardial infarction (AMI) in mice was established by ligating the left anterior descending (LAD) coronary artery. Echocardiography was performed to evaluate mice cardiac function in different groups, including sham, model, NCTS low-dose, NCTS high-dose and Trimetazidine groups. In addition, an in vitro oxygen-glucose deprivation/recovery (OGD/R) injury model on H9C2 cells was established to clarify the effects and regulated energy metabolism mechanism of NCTS. H9C2 cells were divided into four groups, including control, model, NCTS treatment and NCTS+HX531 (RXRα inhibitor) treatment groups. Western Blot was used to detect the expression of RXRα, PPARα, CPT1a, CD36. Immunofluorescence detected the expression of RXRα in myocardial tissue. Adenosine Triphosphate (ATP) were examined by ATP Assay Kit.
RESULTS In this study, NCTS could dose-dependently bind to the RXRα protein with a KD value of 2.5 × 10-5 M, demonstrating that NCTS potentially bound with RXRα as a direct ligand. Echocardiography results indicated that NCTS could protect cardiac function. Compared with model group, NCTS could increases the expression of RXRα and the level of ATP in myocardial tissue (P<0.05). Positive drug Trimetazidine showed similar effects. Simultaneously, similar effects were revalidated in OGD/R-induced H9C2 injury model. NCTS in 2 μmol/L had protective effects. We discovered that compared with the model group, RXRα was significantly increased in the NCTS treatment group (P<0.01), and the co-treat with RXRα antagonist HX531 can eliminate the effect of NCTS. Interestingly, the expression of PPARα, CPT1a and CD36 decreased in model group (P<0.01). After treatment with NCTS, the expression of PPARα, CPT1a and CD36 was notably restored (P<0.05). In the NCTS+HX531 group, the effect of NCTS is also eliminated. These findings suggested that NCTS can effectively activate RXRα expression and protect cardiocytes by regulating RXRα-mediated energy metabolism pathway.
CONCLUSIONS This study demonstrates that NCTS can effectively improve the cardiac function of mice with HF post-AMI and has a protective effect on cardiomyocytes. The mechanism of NCTS is to improve energy metabolism disorders through targeting on RXRα. The present study provides a potential target-drug for clinical treatment of cardiovascular disease.
GW31-e0452
Renyang Tong, Longwei Xu, Yichao Zhao, Jun Pu
Department of Cardiology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University
OBJECTIVES Sepsis and its resultant heart failure, characterized by the dysregulated inflammatory response, are among leading causes of death worldwide. However, no specific approaches have proven clinically effective and efficient in the treatment of septic cardiac dysfunction. Rev-erbα, a unique member of the nuclear receptors (NRs), was recently reported to pivotally participate in inflammation regulation. However, the role of Rev-erbα in sepsis-induced cardiac dysfunction remains unknown.
METHODS (1) Cardiomyocyte-specific Rev-erbα-knockout mice (KO) were established by Shanghai Model Organisms Center, Inc. (2) The mice septic model was established by a dose intraperitoneal injection (i.p.) of Escherichia Coli O55:B5 LPS into 12- to 14-week-old male mice at ZT6 or ZT18 for 6 hours according to previous reports. (3) Polymicrobial sepsis was induced by Cecal ligation and puncture (CLP) in 12- to 14-week-od male mice as described previous. (4) 12 overnight-fasted Bama miniature pigs (30±2 kg) were utilized to establish sepsis pigs model. (5) In vivo cardiac function of each group mice was measured using echocardiography (Vevo 2100, VisualSonic, Toronto, Canada) and left ventricular hemodynamics (AD Instruments, Castle Hill, New South Wales, Australia) 6 hours after LPS injection, as previously described. (6) To assess the extent of myocardial interstitial edema of mice, in vivo CMR was conducted using a 7.0 T small animal MRI system (BioSpec70/20USR; Bruker BioSpin GmbH, Ettlingen, Germany; Software ParaVision 5.1). (7) The related mRNA and proteins expression were respectively determined by RT-PCR, Western blot and Enzyme-linked immunosorbent assay (ELISA). (8) Chromatin immunoprecipitation (ChIP) assays were performed by using SimpleChIP® Plus Enzymatic Chromatin IP Kit (#9005, CST, Beverly, MA) per manufacturer’s protocol. (9) ChIP-seq library preparation and sequencing were performed by RiboBio.
RESULTS (1) Firstly, we found Rev-erbα owned the highest expression abundance among 49 NRs in the murine hearts, and once challenged with LPS, endogenous Rev-erbα level was significantly down-regulated in hearts from septic mice and LPS-treated mouse or human cardiomyocytes. (2) And we noticed that the diurnal oscillation of Rev-erbα expression was associated with the severity of cardiac injury in septic mice. (3) Furthermore, in vivo loss-of-function experiments indicated that Rev-erbα deficiency visibly exacerbated sepsis-induced cardiac dysfunction, inflammatory response and mortality. (4) Meanwhile, the protective role of Rev-erbα was confirmed in the in vitro loss-of- and gain-of-function experiments in the primary murine cardiomyocytes. (5) Additionally, pharmacological activation of Rev-erbα by SR9009 obviously ameliorated the LPS-induced adverse phenotype in both mice and bama miniature pigs. (6) Mechanistic studies revealed that monocarboxylate transporter 8 (MCT8, one of main thyroid hormone transporters) was a direct transcriptional activation target of Rev-erbα via binding with the its promoter region and mediated the Rev-erbα-elicited cardioprotection.
CONCLUSIONS In summary, the present study provides the first evidence that Rev-erbα acts as a novel protective receptor against septic cardiac injury via directly transactivating the TH transporter MCT8. Therefore, targeting the Rev-erbα-MCT8 signaling may represent a promising strategy to alleviate depressed hearts during sepsis.
GW31-e0476
Chi Zhang
Wenzhou Medical University
OBJECTIVES Our previous studies showed that both exogenous and endogenous FGF21 inhibited cardiac apoptosis at the early-stage of type 1 diabetes. Whether FGF21 induces preventive effect on type 2 diabetes-induced cardiomyopathy was investigated in the present study.
METHODS High-fat-diet/streptozotocin-induced type 2 diabetes was established in both wild-type (WT) and FGF21-knockout (FGF21-KO) mice followed by treating with FGF21 for 4 months. Cardiac function, morphological changes, cardiac hypertrophy, fibrosis as well as apoptosis, oxidative stress and inflammation were diagnosed.
RESULTS Diabetic cardiomyopathy (DCM) was diagnosed by significant cardiac dysfunction, remodeling and cardiac lipid accumulation associated with increased apoptosis, inflammation and oxidative stress, which was aggravated in FGF21-KO mice. However, the cardiac damage above was prevented by administration of FGF21. Further studies demonstrated that the metabolic regulating effect of FGF21 is not enough contributing to FGF21-induced significant cardiac protection under diabetic condition. Therefore, other protective mechanisms must exist. The in vivo cardiac damage was mimicked in primary neonatal or adult mouse cardiomyocytes treated by HG/Pal, which was inhibited by FGF21 treatment. Knockdown of AMPKα1/2, AKT2 or NRF2 with their siRNAs revealed that FGF21 protected cardiomyocytes from HG/Pal partially via up-regulating AMPK-AKT2-NRF2-mediated anti-oxidative pathway. Additionally, knockdown of AMPK suppressed fatty acid β-oxidation via inhibition of ACC-CPT-1 pathway. And, inhibition of fatty acid β-oxidation partially blocked FGF21-induced protection in cardiomyocytes. Further in vitro and in vivo studies indicated that FGF21-induced cardiac protection against type 2 diabetes was mainly attributed to lipotoxicity rather than glucose toxicity.
CONCLUSIONS FGF21 functions physiologically and pharmacologically to prevents type 2 diabetic lipotoxicity-induced cardiomyopathy through activation of both AMPK-AKT2-NRF2-mediated anti-oxidative pathway and AMPK-ACC-CPT-1-mediated lipid-lowering effect in the heart.
GW31-e0485
Yunli Shen, Hao Zheng, Xiaobo Yao, Qizheng Lu
Department of Cardiology, Shanghai East Hospital, Tongji University School of Medicine
OBJECTIVES Magnetic iron oxide nanoparticles (MNP) as contrast agents or target carriers have been widely used in in the diagnosis and treatment for cardiovascular diseases. Correspondingly, the myocardial tissue safety of MNP is becoming the bottleneck to seriously restrict its clinical translation. We aimed to investigated whether MNP could aggravate oxidative stress injury in ischemic myocardium and further explored whether a potent antioxidant N-acetylcysteine (NAC) modifying could neutralize the toxicity of MNP.
METHODS NAC modified MNPs are prepared. We injected different nanoparticles in a rat model of ischemia/reperfusion, including MNP, mesoporous silica nanoparticles (MSN), magnetic mesoporous silica nanoparticles (M-MSN) and NAC loaded M-MSN (M-MSN@NAC). The myocardial ROS levels in the peri-infarct zone were detected by DHE 24 hours after the intramyocardial injection. At 2 weeks after intramyocardial injection, the MDA, GSH-Px and GSH were measured using colorimetric method or ELASA. The mitochondrial membrane potential (MMP) and ATP of myocardial cells were respectively detected by JC-1 kit and ATP Test Kit. Myocardial tissue non-heme iron levels were measured using the chromogen method. Myocardial mitochondria at injection sites were observed by transmission electron microscopy. At 4 weeks after injection, the cardiac function was measured by using echocardiography, and histologic analyses of infarct morphology was obtained.
RESULTS Both the MNP group and the M-MSN group showed higher levels of ROS in ischemic myocardium than that in the control group at 24 hours after injection (both P<0.0001). At 2 weeks after injection, compared with the control group, the MNP, M-MSN and M-MSN@NAC groups demonstrated significant increased content of non heme iron in cardiomyocytes (all P<0.01), especially dramatically increased of non heme iron content in mitochondria (all P<0.01). Compared to the control group, both MNP and M-MSN induced the significant increase of MDA (both P<0.01), the marked decrease of GSH (both P<0.01), the inactivation of GPx4 (both P<0.01), the loss of MMP and ATP depletion (all P<0.0001), suggesting that iron oxide nanoparticles could cause mitochondrial lipid peroxidation by inducing mitochondrial iron overload. The level of mitochondrial MDA in M-MSN@NAC group was lower than that in the MNP and M-MSN groups (both P<0.01), but it was still significantly higher than that in the control group (P<0.01). Moreover, the MMP and ATP levels in M-MSN@NAC group were similar to the MNP and M-MSN groups (all P>0.05), implying that NAC could only partly relieved mitochondrial lipid peroxidation induced by MNP or M-MSN. In addition, compared with the control group, the M-MSN with or without NAC modification also could cause serious destruction of mitochondrial structure. At 4 weeks after injection, MNP and M-MSN exaggerated the left ventricular negative remodeling and functional deterioration (all P<0.0001), and M-MSN@NAC exhibited NAC modification could only slightly mitigate the myocardial toxicity induced by iron oxide nanoparticles.
CONCLUSIONS The local application of magnetic iron oxide nanoparticles can aggravate the negative remodeling of ischemic myocardium by induction of mitochondria iron overload to lead to lipid peroxidation. Traditional antioxidant NAC can only mildly reduce the myocardial toxicity.
GW31-e0507
Xiuting Sun, Xiaodong Zhuang, Xinxue Liao
First Affiliated Hospital of Sun Yat-Sen University
OBJECTIVES Doxorubicin (Dox)-induced cardiotoxicity could lead to dilated cardiomyopathy and heart failure, which limited its clinical application. Our previous study reported the protective effects of Klotho against hyperglycemia-induced cardiomyopathy. We investigated whether Klotho alleviated Dox-induced cardiotoxicity, and dissected the underlying mechanism.
METHODS Primary neonatal rat ventricular cardiomyocytes and H9c2 cells were incubated with 5 μM Dox for 24 h with or without Klotho (0.1 mg/mL). Dox-induced cardiotoxicity model was approached in C57BL/6 mice. Both in vitro and in vivo study were conducted in four groups (control, Dox, Klotho and Dox+Klotho group). Cardiac function and serum enzyme activity, apoptosis and mitochondrial dysfunction were measured.
RESULTS Pretreatment with Klotho significantly reduced Dox-induced apoptosis, mitochondrial fission and inflammation in cardiomyocytes. In Dox-treated mice, Klotho also suppressed cardiac cell death and inflammatory response, improved cardic function. The expression of Dynamin-related protein 1 (Drp1) was also increased after Dox-treatment. Furthermore, overexpression of Drp1 in cardiomyocytes increased Dox-induced heart injury which could also be attenuated by Klotho.
CONCLUSIONS This study demonstrated that Klotho alleviated Dox-induced cardiotoxicity by reducing apoptosis and mitochondrial fission though down-regulating Drp1 expression. Our findings highlight new targets for the therapy of Dox-induced cardiomyopathy.
GW31-e0509
Yue Guo1,2, Xingfeng Xu1,2, Huimin Zhou1, Xiaodong Zhuang1, Xinxue Liao1,2
1Department of Cardiology, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, P.R. China
2NHC Key Laboratory of Assisted Circulation (Sun Yat-sen University), Guangzhou, Guangdong 510080, P.R. China
OBJECTIVES Excessive mitochondrial fission plays a critical role in the pathogenesis of diabetic cardiomyopathy (DCM). Our previous study demonstrated that anti-aging protein Klotho attenuated hyperglycemia-induced myocardial injury and cardiac dysfunction. Silent information regulator 1 (SIRT1) is an NAD+ dependent protein deacetylase involved in the cardioprotective effects of Klotho. However, whether SIRT1 signaling is involved in the regulatory effect of Klotho on mitochondrial fission has not been elucidated. Therefore, the purpose of this study were (i) to explore whether Klotho attenuates diabetes-induced cardiac dysfunction by inhibiting mitochondrial fission; (ii) if so, to determine whether Klotho prevents mitochondrial fission via activation of SIRT1 signaling.
METHODS Neonatal cardiomyocytes were exposed to 33 mM glucose in the presence or absence of Klotho (400 pM). Moreover, we established a diabetic mouse model to assess the protective effect of Klotho. Control and streptozotocin-induced diabetic mice were untreated or treated with Klotho (Intraperitoneal injection of Klotho at 0.01 mg/kg per 48 h) 1 weeks after diabetes induction and assessed 3 months afterward.
RESULTS Klotho prevented diabetes-induced myocardial dysfunction by inhibiting dynamin-related protein 1 (Drp1)-mediated mitochondrial fission. Klotho inhibited Drp1 expression, reduced oxidative stress, decreased cardiomyocyte apoptosis, improved mitochondrial function and cardiac function in diabetic mice, but not in SIRT1-/- diabetic mice. In addition, we found Klotho treatment increased the expression of SIRT1 and PGC-1α and inhibited Drp1-mediated mitochondrial fission and mitochondria-derived superoxide production in neonatal cardiomyocytes. Moreover, inhibition of SIRT1 or PGC-1α (by siRNA) canceled inhibitory effects of Klotho on Drp1 expression and mitochondrial fission. These data indicated that Klotho exerted its cardioprotective effects by reducing Drp1-mediated mitochondrial fission in a SIRT1/PGC-1α-dependent manner. Chromatin immunoprecipitation analysis further showed that PGC-1α negatively regulated Drp1 expression by binding to its promoter. Drp1 inhibitor mdivi-1 alleviated oxidative stress, mitochondrial dysfunction and myocardial dysfunction in diabetic mice.
CONCLUSIONS This work demonstrated that anti-aging protein Klotho prevented diabetes-induced cardiac dysfunction by inhibiting mitochondrial fission via SIRT1/PGC-1α/Drp1 pathway.
GW31-e0510
Xiuting Sun, Xiaofong Zhuang, Xinxue Liao
First Affiliated Hospital of Sun Yat-Sen University
OBJECTIVES Pyroptosis played a crucial role in the development of diabetic endothelial injury. Endothelial microparticles (EMPs) served as potential drug delivery carriers that can carry non-coding RNAs which activated endothelial cells. Studying the function of miRNAs in EMPs will provide new perspectives for the clinical diagnosis and prognostic assessment of endothelial functional impairment.
METHODS Male db/db+ mice were randomly divided into: control, DMEMP, Non-DMEMP, DMEMP+miR-26a, DMEMP+miR-scr groups. EMP was administered intravenously 3 times a week for 4 weeks. EMP was administered by tail vein with an insulin needle three times a week for 4 weeks; 0.1 mL of 5 nmol miR-26a Agomir and miR-scr was administered by tail vein twice a week for 4 weeks. Three db/db+ and three db/m+ mice were selected, venous blood was collected, centrifuged to obtain EMP, high-throughput sequencing to screen for differential gene expression, and luciferase assay to verify miRNA target genes. Three db/db+ and three db/m+ mice were selected, high-throughput sequencing to screen for differential gene expressions in EMPs, luciferase assay to verify miRNA target genes. Thoracic aorta of mice was tested for vasodilatory function, and the aorta of each group of mice was stained with Hematoxylin and eosin (HE), and the aorta of mice was tested for unc-51-like kinase 1 (ULK1), NLRP3 inflammatory small body and cytokine-related protein expression by Western blot. Immunofluorescence detection of NLRP3 and Caspase 1 expression in the aorta of mice free of each group. Different EMPs were co-cultured with HUVEC, HUVEC migration and proliferation were observed. miR-26a was transfected with HUVEC to detect cell proliferation and migration function, Western blot to detect hydrolytic cleavage of NLRP3, Caspase-1, ULK1 expression, immunofluorescence to detect NLRP3 and Caspase 1 co-localization, and cell coke death assay kit to detect the number of cells with Active caspase-1+/7-AAD+; YVAD or MCC 950 pretreatment with HUVEC, pyroptosis assay kit was used to the number of cells with active Caspase-1+/7-AAD+; Western blotting was used to detect hydrolytic cleavage changes in Caspase-1 and GSDMD.
RESULTS Compared with the control group, DMEMP damaged the repair function of aortic endothelium, but no significant reduction in endothelial repair function was seen in the Non-DMEMP group. miR-26a expression was significantly increased in db/db+ mouse EMPs. The luciferase report validated ULK1 as a target gene for miR-26a. In EMPmiR-26a-treated mice, adhesion of aortic endothelial cells was further disrupted. DMEMP-treated mice and HUVEC showed increased co-localization of Caspase 1 with NLRP3 and increased TUNEL co-localization with Caspase 1; Western blot detected hydrolytic cleavage of Caspase-1, GSDMD, IL-1β and IL-18. After transfection with HUVEC, miR-26a mimic suppressed ULK1 expression, inhibiting cell proliferation and migration. miR-26a mimic transfection of HUVEC revealed increased co-localization of Caspase 1 with NLRP3 and increased TUNEL co-transfection with Caspase 1. Protein expression of Caspase-1, GSDMD, IL-1β, and IL-18, flow cytometry detection of increased cell pyroptosis. Effects caused by DMEMP-miR-26a were significantly inhibited by pretreatment with YVAD (Caspase-1 inhibitor), MCC950 (NLRP3 inflammatory small body inhibitor).
CONCLUSIONS Diabetic EMPs can lead to endothelial impairment by exerting endothelial damage by carrying miR-26a, which acts through the target protein ULK1. DMEMP-carrying miR-26a promotes the onset of cell pyroptosis death by targeting downregulation of ULK1 expression, resulting in endothelial damage.
GW31-e0512
Chendie Yang, Xiaoqun Wang
Rui Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, P.R. China
OBJECTIVES Atherosclerotic lesions preferentially occur at branch points of arterial trees where the blood flow is disturbed. Disturbed flow increases endothelial permeability, vascular barrier dysfunction, and finally the development of atherosclerosis. CTRP1, a member of C1q/TNF related protein (CTRP) family, is a novel secreted glycoprotein and its biological functions are largely undefined.
METHODS Endothelial permeability was determined by Miles assay in vivo and F-actin staining in vitro. CTRP1 knockout mice were also established to further determine the effect of CTRP1 on vascular barrier function.
RESULTS We found CTRP1 expression was significantly increased by disturbed flow in endothelial cells, thereby promoting cytoskeleton assembly and the formation of paracellular holes. Inhibition of CTRP1 either by specific siRNA notably attenuated disturbed flow-dependent endothelial hyperpermeability. Next, CTRP1 knockout mice were established. Miles assay demonstrated impaired vascular barrier function in mice injected with recombinant CTRP1 proteins, but was greatly improved in CTRP1 knockout mice. Finally, we found leukocyte trans-endothelial migration and atherosclerotic progression was markedly attentuated either in CTRP1 knockout mice or by inhibition of CTRP1 with intravenous injection of specific neutralizing antibodies.
CONCLUSIONS CTRP1 is a mechano-sensitive proinflammatory factor that mediates disturbed flow-induced vascular barrier dysfunction. Inhibition of CTRP1 may inhibit the pathogenesis of atherosclerosis at early stage.
GW31-e0513
Chendie Yang, Xiaoqun Wang
Ruijin Hospital, Shanghai Jiaotong University School of Medicine
OBJECTIVES C1q/TNF-related protein (CTRP) 1 was initially identified as a paralog of adiponectin based on the similarity in C1q domain of these two proteins. Previously, we showed that CTRP1 promotes the development of atherosclerosis by increasing endothelial adhesiveness. Here, we sought to investigate whether CTRP1 also influences vascular dilatory functions.
METHODS Vascular dilatory responsiveness was compared by intravital microscopy of cremaster arterioles between CTRP1 transgenic (Tg-CTRP1), CTRP1 knockout (CTRP1 KO) and C57 wild type (WT) control mice. In situ production of reactive oxygen species (ROS) was determined by dihydroethidium (DHE) staining.
RESULTS We found a dramatic impairment of endothelium-dependent arteriolar dilation in Tg-CTRP1 mice, whereas vasodilation was markedly enhanced in CTRP1 KO mice as compared to WT controls. Meanwhile, elevated production of reactive oxygen species (ROS) was detected in the vascular wall of Tg-CTRP1 animals. In cultured endothelial cells, CTRP1 stimulation resulted in reduced nitric oxide (NO) bioavailability both in the cell lysates and conditioned media. Furthermore, we found that arginase 1 was significantly increased by CTRP1 in a dose-dependent fashion, thereby leading to endothelial nitric oxide synthase (eNOS)-uncoupling and reactive oxygen species generation. Inhibition of arginase activity by synthetic chemicals markedly improved CTRP1-dependent vasodilatory dysfunctions.
CONCLUSIONS These data define the essential role of CTRP1 in mediating vasodilatory dysfunctions, as well as propose a novel mechanism that increased arginase activity by CTRP1 leads to uncoupling of eNOS homodimers, thereby limiting NO biosynthesis and amplifying ROS production.
GW31-e0514
Chendie Yang, Xiaoqun Wang
Ruijin Hospital, Shanghai Jiaotong University School of Medicine
OBJECTIVES Increased transcytosis of low-density lipoprotein (LDL) across the endothelium and oxidation of LDL deposited within the subendothelial space are crucial early events in atherogenesis. C1q/TNF-related protein (CTRP) 5 is a novel secreted glycoprotein and its biological functions are largely undefined.
METHODS In this study, we analyzed CTRP5 levels in sera of patients with coronary artery disease (CAD, n=288) and non-CAD controls (n=264). In this study, we analyzed CTRP5 levels in sera of patients with coronary artery disease (CAD, n=288) and non-CAD controls (n=264). The role of CTRP5 in LDL transcytosis and oxidative modification was investigated in vivo and in vitro.
RESULTS We found CTRP5 serum levels were higher in patients with than without CAD (247.26±61.71 vs. 167.81±68.08 ng/mL, P<0.001), and were positively correlated to the number of diseased vessels (Spearman’s r=0.611, P<0.001). Increased expression of CTRP5 was detected in human coronary endarterectomy specimens as compared to non-atherosclerotic arteries. Immunofluorescence showed that CTRP5 was predominantly localized in endothelium and macrophages in human atherosclerotic lesions. In vivo and in vitro experiments demonstrated that CTRP5 promoted transcytosis of LDL across endothelial monolayers, as well as the oxidative modification of LDL in endothelial cells. Inhibition of CTRP5 with a neutralizing antibody dramatically attenuated the deposition of oxidized lipids in the aortic wall of ApoE-/- mice. Mechanistically, we found that CTRP5 up-regulated 12/15-lipoxygenase (LOX), a key enzyme in mediating LDL trafficking and oxidation, through STAT6 signaling. Genetic or pharmacological inhibition of 12/15-LOX dramatically attenuated the deposition of oxidized LDL in subendothelial space and the development of atherosclerosis.
CONCLUSIONS These data indicate that CTPR5 is a novel pro-atherogenic cytokine and promotes transcytosis and oxidation of LDL in endothelium through up-regulating 12/15-LOX.
GW31-e0521
Xiaoqun Wang, Lin Lu
Rui Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, P.R. China
OBJECTIVES Atherosclerosis is predisposed at bifurcations, branch points and arterial curvatures where the blood flow is disturbed. Deposition and oxidative modification of low-density lipoprotein (LDL) in the subendothelial intima is an early event in the pathogenesis of atherosclerosis. However, the effect of disturbed flow (d-flow) on LDL oxidation and the underlying mechanisms remain unclear. In this study, we investigated the role of 12/15-lipoxygenase (LOX) in LDL oxidation in endothelial cells (EC) and the development of atherosclerosis at d-flow sites.
METHODS En face immunofluorescence was performed to analyze the expression of 12/15-LOX in EC under exposure to different flow patterns. Protein expression and LDL oxidation was studied in vitro by using 2 different flow apparatuses. Animal model of d-flow was established by partial ligation of the carotid artery.
RESULTS En face staining exhibited a significant increase in 12/15-LOX protein expression in EC in areas exposed to d-flow than those exposed to steady flow (s-flow). Carotid partial ligation in ApoE knockout mice led to substantially increased deposition of oxidized LDL in the subendothelial intima and formation of atherosclerotic plaques in the carotid artery, whereas these detrimental effects by d-flow were markedly attenuated in ApoE/12/15-LOX double knockout mice. In cultured EC, d-flow generated by a reversal flow pump markedly promoted the expression of 12/15-LOX and translocation of the protein onto the cell membrane. Inhibition of 12/15-LOX in EC, either by knockdown with its specific siRNA or a pharmacological inhibitor, evidently suppressed LDL oxidation in response to d-flow. Mechanically, we found d-flow induced the expression of 12/15-LOX by activating a specific responsive element in the 12/15-LOX promoter through recruiting a shear stress-sensitive transcriptional factor SREBP2. Chromatin immunoprecipitation further confirmed the interaction of SREBP2 with the promoter of 12/15-LOX upon d-flow exposure.
CONCLUSIONS These data define an essential role of 12/15-LOX in promoting the pathogenesis of atherosclerosis under d-flow by increasing LDL oxidation in EC through SREBP2 signaling.
GW31-e0523
Xia Fan, Xiaoqing Yan, Yi Tan
Wenzhou Medical University
OBJECTIVES Diabetic vascular disease is one of most serious complications of diabetes mellitus. Promoting angiogenesis is an important strategy for the treatment of diabetic vascular disease. Endothelial progenitor cell (EPC) plays a key role in angiogenesis. The number of EPC is decreased and the function of EPC impaired in diabetic conditions. Increasing the number of EPC and/or improving EPC function can effectively promote diabetic ischemic angiogenesis. Fibroblast growth factor 21 (FGF21) is a member of the fibroblast growth factor family that plays a key role in regulating glucose and lipid metabolism and has the potential to treat diabetes mellitus and its complications. However, its role in diabetic vascular disease is largely unknown. This study explored the effect of FGF21 on diabetic ischemic angiogenesis and the underlying mechanism.
METHODS C57BL/6J mice received 5 doses STZ injection to induce type 1 diabetes mellitus. Hind limb ischemia (HLI) model was established by unilateral femoral aortic ligation, and 200 ng/mL FGF21 or equal volume of vehicle was administered by intraperitoneal injection every day. Laser Doppler assay was used to evaluate the blood perfusion of ischemic hind limb at day 0, 3, 7, 14, 21 and 28 days after surgery. After the last Laser Doppler assay, mice were sacrificed and the gastrocnemius muscle tissues of ischemic hind limb were collected. The expression of CD31 was detected by immunofluorescence staining to valuate the number of neovascularization in ischemic hind limbs. EPC was extracted from human umbilical cord blood and incubated in culture medium containing 33 mM glucose for 24 h to mimic in vivo diabetic condition, along with or without 200 ng/mL FGF21. The migration capability of EPC was investigated by scratch assay, and the angiogenic ability of EPC was investigated by matrix tube-like structure formation assay to evaluate the protective effect of FGF21 on the migration and tubing function of EPC treated with high glucose. The content of NAD+ in cell lysate was detected by the kit, and the expression of proteins of related pathways was detected by Western Blot.
RESULTS FGF21 could enhance blood perfusion in ischemic site of type 1 diabetic mice, but had no significant effect on blood sugar and body weight. The expression of CD31 in ischemic gastrocnemius muscle of mice treated with fibroblast growth factor 21 was up-regulated, indicating enhanced neovascularization. In vitro study also showed that both the receptor FGFR1 and co-receptor beta-klotho (KLB) were expressed on the cell surface of EPC. FGF21 can also stimulate AKT phosphorylation in EPC. Moreover, the migration and tubing ability of EPC were impaired by high glucose, while FGF21 can ameliorate this process. Meanwhile, the content of NAD+ in EPC treated with high glucose decreased, and it increased with the administration of fibroblast growth factor 21. Moreover, NAD+ supplementation can improve EPC migration and tubular damage caused by high glucose.
CONCLUSIONS FGF21 can effectively promote the recovery of blood flow in HLI tissue of type 1 mice. FGF21 may improve the angiogenesis ability of EPC treated with high glucose by increasing the content of NAD+ in EPC cells.
GW31-e0524
Shaohua Wang, Sixu Chen, Yangxin Chen
Department of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University
OBJECTIVES Cardiac mitophagy and mitochondrial respiratory function are altered during lipotoxicity but the underline mechanism are not largely unknown.
METHODS Palmitic acid and ob/ob mice were used to induce cardiac lipotoxicity in vitro and in vivo, respectively. Mitochondrial function was evaluated by measuring mitochondrial mass, respiratory complex expression and respiratory capacity. Mitochondrial biogenesis was determined by assessing related transcription factors (PPARγ coactivator 1α, PGC-1α and mitochondrial transcription factor A, TFAM). Mitophagy was determined by transmission electron microscopy (TEM) and fluorescence indicators (immunofluorescence co-staining of LC3 and Tomm20, mitoTimer and mtKemia). The crucial mitophagy pathways were identified and small molecule RNA interference against the target gene was used to assess the role of mitophagy in lipotoxic cardimomyocytes. Potentially critical involved long-coding RNAs were screened by analyzing high throughput data, function and mechanism of which on mitophagy were explored. The underlined mechanism regulating the expressions of the target lncRNA during lipotoxicity were also addressed.
RESULTS Lipotoxicity resulted in a substantial decrease of mitochondrial number and volume, along with a reduction of respiratory capacity. Lipotoxicity increased PGC-1α and TFAM protein levels and promoted mitophagy, as indicated by TEM and fluorescence indicators. Fundc1 and Bnip/Nix were not changed significantly upon lipotoxicity while obvious up-regulation of Pink1 and Parkin were observed. Moreover, ubiquitination of mitochondrial membrane proteins were found. Knocking down of Parkin attenuated palmitic acid-induced mitophagy and increased respiratory capacity. Analysis of the lipid accumulation high throughput dataset (GEO Number: GSE27975) found that lncRNA H19 expression was mostly altered among the genes with human and rat homologous sequences. Forced expression of H19 dramatically reduced Pink1 expression, along with a decrease of mitophagy and a restoration of respiratory capacity. Pink1 mRNA was not changed by palmitic acid while RNA antisense purification (RAP, using H19 probe)-PCR revealed a combination of H19 and Pink1 mRNA. H19 was predicted to bind to eukaryotic translation initiation factor 4A3 (eIF4a3) but result from H19 RAP-mass spectrometry found eIFA2 rather than eIFA3 and RAP-Western blot showed much more eIF4a2 than eIF4A3 in abundance. RNA Immunoprecipitation using eIF4a2 antibody demonstrated the combination of eIF4a2 protein, H19 and Pink1 mRNA. Methylation levels of H19 promoter sequence were significantly increased by palmitic acid. Expression and activity of methylase Dnmt3b were up-regulated while those of Dnmt1 and Dnmt3a were suppressed. Chromatin-immunoprecipitation using Dnmt3b antibody yield a significantly enriched sequences around the putative methylation regions inside H19 promoter.
CONCLUSIONS Down-regulation of H19 played a crucial role in lipotoxicity-induced cardiac mitochondrial capacity impairment via promoting mitochondria mitophagy. H19, the expression of which was decreased by promoter methylation under lipotoxic circumstances, inhibited excessive mitophagy by imitating Pink1 mRNA translation.
GW31-e0531
Yujie Xing, Ying Lv, Dong Liang, Xiaoxiang Liu, Junkui Wang
First Department of Cardiology, Shannxi Provincial People’s Hospital, Xi’an, Shaanxi 710068, People’s Republic of China
OBJECTIVES To investigate the role of Wnt/β-catenin signaling pathway in heart failure of rats with selenium deficiency.
METHODS Forty rats were randomly divided into the control group and the selenium deficiency group, with twenty rats in each group. Rats in the control group were fed with the standard feed, and rats in the selenium deficiency group were fed with selenium deficiency diet. All rats were fed for sixteen weeks. Then the blood selenium and BNP level of the rats were measured by the eyeball extraction, the cardiac function of the rats was detected by echocardiography, and the expression level of Wnt and β-catenin protein were detected by Western Blotting.
RESULTS The blood selenium level in selenium deficiency group was significantly lower than that in control group, while BNP level in selenium deficiency group was significantly higher than that in control group. Echocardiography showed that the heart function of the selenium deficiency group was lower than that of the control group. Western Blot showed that the expression level of Wnt and β-catenin in selenium deficiency group was higher than that in control group.
CONCLUSIONS Selenium deficiency can cause heart failure of rats, and its mechanism may be related to the activation of Wnt/β-catenin signaling pathway.
GW31-e0544
Yingyu Xie1,2, Zihan Fang3, Mingyang Wang1, Yanan Wang1, Junping Zhang2
1Tianjin University of Traditional Chinese Medicine
2First Teaching Hospital of Tianjin University of Traditional Chinese Medicine
3China National Center for Biotuchnology Development
OBJECTIVES To observe the intervention effect of Yiqi Huoxue recipe (YQHX) on rats with chronic heart failure, in order to explore mitochondrial function mechanism.
METHODS The left anterior descending coronary artery ligation was performed to construct the chronic heart failure (CHF) rat model. Among 64 male SD rats, 16 were randomly selected as the sham operation group. After modeling, they were randomly divided into model group, captopril group and YQHX group. After 8 weeks of intervention, cardiac tissues were collected, body mass and heart mass were weighed, and echocardiography were performed to detect the changes in cardiac structure, and detection of NT-probNP to determine the degree of heart failure in rats. Masson staining was performed to determine the myocardial interstitial collagen volume fraction. Transmission electron microscopy was used to observe the structure and morphology of mitochondria in myocardial tissue. Western blot was used to detect the expression levels of mitochondrial fusion protein optic atrophy 1 (Opa1) and cleavage protein dynamic-related protein 1 (Drpl). The quantitative Real-time PCR was applied to detect the expressions of PINK1/Parkin pathway-related factors.
RESULTS Compared with the sham group, the left ventricular wall of the model group was significantly thickened (P<0.05), the cardiac cavity was significantly enlarged, NT-proBNP was increased (P<0.01), and the content of collagen in the myocardial interstitium was increased (P<0.01). The expression level of Opal decreased, the expression level of Drp1 increased (P<0.05), the mRNA expression level of PINK1 and Parkin increased (P<0.01). Compared with the model group, YQHX group can reduce ventricular wall thickening, heart chamber enlargement, myocardial interstitial collagen content, up-regulate the low expression of Opa1, but down-regulate the high expressions of Drpl, PINK1, Parkin (P<0.05, P<0.01).
CONCLUSIONS YQHX can effectively improve mitochondrial function in rats with CHF. The mechanism may be related to the regulation of PINK1/Parkin and the maintenance of mitochondrial dynamic stability.
GW31-e0555
Yanhua Xuan1, Shuangshuang Liu2, Zhijun Sun2
1Department of Cardiology Medicine, Liaoning Provincial People’s Hospital
2Department of Cardiology Medicine, Shengjing Hospital of China Medical University
OBJECTIVES Previous studies have reported that short-term vagus nerve stimulation (VNS) improves cardiac function in rats with chronic heart failure (CHF). The molecular mechanisms are unclear. The potential effect of microRNA (miR)-126 in apoptosis of short-term VNS was examined.
METHODS A total of 3 weeks after inducing CHF, the rats were divided into three groups: Sham stimulation in sham operated rats, sham stimulation in CHF rats (CHF-SS), and treated with VNS in CHF rats (CHF-VNS). The right vagus nerve of the neck was stimulated for 72 h in CHF rats with rectangular pulses of 40 ms duration at 1 Hz and 5 V. miR-126 was focused on, which exhibited differential expression in the miRNA microarray analysis of CHF rats, and the effects of VNS on apoptosis were examined.
RESULTS It was verified that the expression level of miR-126 in the CHF-VNS group was increased, and the expression was reduced in the CHF-SS group. Furthermore, mimics or inhibitor of miR-126 was transfected into H9c2 to investigate its function on apoptosis. B-cell lymphoma 2 (Bcl-2) was confirmed a target of miR-126 through a dual luciferase reporter assay and western blotting. It was demonstrated that upregulated miR-126 decreased apoptosis in H9c2 cells. The apoptosis-associated proteins were further detected in H9c2 cells and rat tissue. The mRNA and protein expression levels of caspase-3 and Bcl-2-associated X protein were decreased in the CHF-VNS group, the expression of Bcl-2 were increased. The results were consistent with the in vitro study in the miR-126 inhibitor group.
CONCLUSIONS The present study demonstrated that short-term VNS decreased apoptosis by upregulating miR-126 in rats with CHF. Therefore, the results of the present study provide basic evidence for short-term VNS in the clinical treatment of CHF.
GW31-e0561
Hanwen Zhang, Andi Xu, Qi Chen
Nanjing Medical University
OBJECTIVES Doxorubicin-induced cardiomyopathy (DiCM) is a primary cause of heart failure and mortality in cancer patients, in which macrophage-orchestrated inflammation serves as an essential pathological mechanism. However, the specific roles of tissue-resident and monocyte-derived macrophages in DiCM remain poorly understood. Our objective is to uncover the origins, phenotypes, and functions of proliferative cardiac resident macrophages and mechanistic insights into the self-maintenance of cardiac macrophage during DiCM progression.
METHODS Mice were administrated with doxorubicin to induce cardiomyopathy. Dynamic changes of resident and monocyte-derived macrophages were examined by lineage tracing, parabiosis, and bone marrow transplantation. We then conducted RNA-sequencing (RNA-seq) experiments to examine the global gene expression profile to dissect the regulatory mechanism of resident reparative macrophage proliferation in the DiCM heart. Both global or macrophage-specific SR-A1 knockout mice was used to confirm the role of SR-A1 in modulating resident reparative macrophage proliferation and DiCM progression. To further address the role of the SR-A1-c-Myc pathway in DiCM pathogenesis, the lentivirus to silence or overexpress macrophage c-Myc was produced using the SP-C1 promoter.
RESULTS We found that the monocyte-derived macrophages primarily exhibited a pro-inflammatory phenotype that dominated the whole DiCM pathological process and impaired cardiac function. In contrast, cardiac resident macrophages were vulnerable to doxorubicin insult. The survived resident macrophages exhibited enhanced proliferation and conferred a reparative role. Global or myeloid specifically ablation of class A1 scavenger receptor (SR-A1) inhibited proliferation of cardiac resident reparative macrophages and therefore exacerbated cardiomyopathy in DiCM mice. Importantly, the detrimental effect of macrophage SR-A1 deficiency was confirmed by the transplantation of bone marrow. At the mechanistic level, we show that c-Myc, a key transcriptional factor for the SR-A1-P38-SIRT1 pathway, mediated the effect of SR-A1 in reparative macrophage proliferation in DiCM.
CONCLUSIONS The SR-A1-c-Myc axis may represent a promising target to treat DiCM through augmentation of cardiac resident reparative macrophage proliferation.
GW31-e0562
Yafei Li, Tianwen Wei, Yi Fan, Hao Wang, Liansheng Wang
Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University
OBJECTIVES Neonatal heart maintains entire regeneration capacity in a transient regeneration window, but the adult heart loses this function. This process involves numerous core hubs dysregulation of expression and activity that adversely affect the myocardium regeneration ability. Present study screened a functional kinase of serine/threonine-protein kinase 3 (SGK3) in cardiac regeneration after neonatal myocardial infarction (MI) using quantitative phosphoproteomics.
METHODS We used quantitative phosphoproteomics data of infarct border zone in newborn heart after MI to identify regeneration related kinases. Gain- and loss-of-function experiments were performed to determine the effect of SGK3 in cardiomyocyte (CM) proliferation and cardiac repair after apical resection (AR) or MI. Pull-down assay and co-immunoprecipitation (Co-IP) experiments were conducted to investigate the direct binding target proteins of SGK3.
RESULTS SGK3 protein expression was highly expressed at postnatal day 1 (P1), reduced at postnatal day 7 (P7) until adult. In vitro, CM proliferation ratio was elevated by SGK3 overexpression, while it was decreased by knockdown of SGK3. In vivo, inhibition of SGK3 shortened the time window of cardiac regeneration after AR in neonatal mice, and overexpression of SGK3 significantly promoted CM proliferation and cardiac repair after MI. Mechanistically, SGK3 could be directly combined with and activated by cyclin dependent kinase 9 (CDK9). Inhibition of CDK9 partially abolished the effect of SGK3 on CM proliferation. Moreover, SGK3 could repress GSK-3β activity and increase β-catenin expression.
CONCLUSIONS Our study revealed a key role of SGK3 in cardiac regeneration following AR or MI injury, which may reopen a novel therapeutic avenue for MI.
GW31-e0568
Yu Ning1,2, Wanwan Wen2, Yunxiao Yang2, Yifan Jia2, Mengling Huang2, Guihao Chen1, Yuejin Yang1, Ming Zhang2
1State Key Laboratory of Cardiovascular Diseases, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College
2Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University
OBJECTIVES Increasing evidence suggests that hypothyroidism aggravates atherosclerosis. Macrophages apoptosis plays a significant role in the formation and progression of atherosclerotic plaques. No known empirical research has focused on exploring the effect of thyroid hormones on macrophage apoptosis in atherosclerotic plaques.
METHODS Mouse RAW264.7 leukemic monocyte-macrophage cells were treated with 100 μg/mL oxidized low density lipoprotein (oxLDL) to elicit foam cell formation. Cells were also incubated with different concentrations of thyroid hormones (T3: 0.38, 0.75, 1.00 μg/L; T4: 25, 50, 100 nmol/L) to study their effects on functions of macrophage foam cells. Small interfering RNA (siRNA) was used to knock down the expression of thyroid hormone receptor alpha1 (TRα1) in RAW264.7. Then the cells survival, oxidative stress and apoptosis were tested.
RESULTS MTT assay revealed that T3 and T4 concentration-dependently decreased the cell proliferation inhibition rates of macrophage foam cells. Thyroid hormones protected RAW264.7 from reactive oxygen species generation induced by oxLDL in a concentration-dependent way, increasing expression of antioxidant enzymes (Sod1, Sod2, Catalase, Gpx1) and decreasing expression of NADPH oxidase subunits (gp91phox, p22phox, p47phox, p67phox). Thyroid hormones concentration-dependently attenuated apoptosis rates and expression of apoptosis-related proteins cleaved Caspase-3 and cleaved Caspase-9 of RAW264.7 induced by oxLDL. After knocking down the expression of thyroid hormone receptor alpha1 (TRα1), the antiapoptotic effects of T3 and T4 were markedly weakened.
CONCLUSIONS Thyroid hormones concentration-dependently promote macrophage foam cells survival and inhibit cell apoptosis. This may be ascribed to the antioxidant effect of thyroid hormones and the antiapoptotic effect of TRα1.
GW31-e0574
Jiuchang Zhong, Chen Fang, Ran Miao, Ying Liu, Juanjuan Song, Jiawei Song, Kun Zuo, Ying Dong, Xinchun Yang
Heart Center and Beijing Key Laboratory of Hypertension, Beijing Chaoyang Hospital Affiliated to Capital Medical University, Beijing 100020, China
OBJECTIVES Activation of the renin-angiotensin system (RAS) and increased angiotensin (Ang) II levels have been implicated in adverse vascular remodeling and progression to hypertension. APLN mediates important physiological effects in the vasculature and is an important pharmacological target while blockade of the RAS and Ang II signaling improves vascular dysfunction; reduce hypertension and cardiovascular events in patients. Adventitial fibroblasts (AFs) are the most abundant cell type in vascular adventitia and play a crucial role in the control of vascular function. Fibroblast growth factor 21 (FGF21) functions as a novel endocrine factor involved in the regulation of glucose, lipid and energy metabolism and has been shown to exert protective functions in hypertension and vascular dysfunction in Ang II-induced hypertensive mice by activation of the angiotensin-converting enzyme 2 (ACE2)/Ang-(1-7) axis. However, little was elucidated upon pathophysiology significance and underlying mechanisms of FGF21 in the AFs.
METHODS Here, we used APLN deficient (APLN-/y) mice and cultured rat AFs to evaluate the influences of recombinant human FGF21 (rFGF21) and Pyr1-Apelin-13 on Ang II-mediated actions by real-time RT-PCR, Western blotting, TUNEL and dihydroethidium fluorescence staining, respectively.
RESULTS The 12-month aged APLN-/y mice developed vascular hypertrophy and dysfunction with reduced ACE2 and FGF21 levels. Ang II infusion potentiated oxidative stress, apoptosis, and pyroptosis in young APLN-/y aorta leading to exacerbation of vascular dysfunction. In cultured rat aortic AFs, stimulation of Ang II led to a marked decrease in FGF21 levels and increases in cellular proliferation, migration and transformation, which were improved by rFGF21 and Pyr1-apelin-13, respectively. Furthermore, pretreatement with rFGF21 strikingly prevented Ang II-induced promotion in ROS generation and levels of proinflammatory factor NF-κB and IL-1β. Notably, administration of rFGF21 and Pyr1-apelin-13 remarkably rescued Ang II-mediated pyroptosis and apoptosis in rat AFs by augmenting of the Bcl-2/Bax ratio and phosphorylated levels of eNOS and preventing expression of pyroptosis related protein caspase-1 and phosphorylated levels of ERK1/2.
CONCLUSIONS In summary, FGF21 and APLN-APLNR pathway are negative regulators of Ang II-mediated adverse vascular remodeling and adventitial dysfunction. We showed that loss of APLN leads to reduction of ACE2 and FGF21 and increased oxidative stress, apoptosis and vascular dysfunction in response to Ang II. Conversely, FGF21 and pyr1-apelin-13 play important roles in the Ang II-mediated oxidative stress, pyroptosis and apoptosis in rat aortic adventitial fibroblasts by modulating the eNOS/ERK signaling. Thus, targeting the FGF21 and APLN pathway represents a novel therapeutic approach against adventitial dysfunction and vascular disorders. This research was supported by the General Program and the National Major Research Plan Training Project of National Natural Science Foundation of China (81770253 and 91849111).
GW31-e0575
Jiuchang Zhong1, Mei Yang1, Juanjuan Song1, Xiaoyan Liu1, Jing Li1, Juan Wang1, Jun Cai2, Xinchun Yang1, Guangzhen Zhong1
1Heart Center and Beijing Key Laboratory of Hypertension, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China
2Hypertension Center, Fuwai Hospital, National Center for Cardiovascular Diseases of China, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100037, China
OBJECTIVES Cardiac remodeling is characterized by ventricular hypertrophy and myocardial fibrosis, which are the main causes of heart failure. MicroRNAs (miRNAs) are a class of endogenous small non-coding RNA molecules and have emerged as crucial regulators of myocardial fibrosis, remodeling and cardiofibroblasts (CFs) dysfunction. We previously found that miRNA-122-5p is significantly upregulated in a rat model of cardiac hypertrophy and dysfunction induced by pressure overload in association with downregulated levels of APLN. However, the exact roles and underlying mechanisms of miRNA-122-5p in the Ang II-induced cardiac fibrosis and CFs dysfunction are still unclear.
METHODS Here, we evaluate roles of miRNA-122-5p in angiotensin (Ang) II-mediated hypertensive rats and rat CFs by real-time RT-PCR, Western blotting, TUNEL, Masson staining and dihydroethidium fluorescence staining, respectively. The Sprague-Dawley rats were randomized to Ang II infusion with an osmotic minipump and pretreated with recombinant adeno-associated viral vector (AAV)-miRNA-122-5p (rAAV-miRNA-122-5p) or rAAV-GFP for 4 weeks.
RESULTS Ang II infusion triggered promotion of myocardial fibrosis and apoptosis and reduction levels of apelin, p-AMPK and LCBII in hypertensive rats, which were exacerbated by rAAV-miRNA-122-5p treatment. In cultured rat CFs, stimulation of Ang II resulted in a marked decrease in autophagy and increases in apoptosis, proliferation and superoxide generation, which were prevented by pretreated with miRNA-122-5p inhibitor. Furthermore, Pyr1-apelin-13 abolished Ang II-induced loss of autophagy and promotion of apoptosis and oxidant injury in rat CFs linked with reduced levels of P62 and enhanced levels of beclin-1. Notably, miRNA-122-5p mimics significantly reversed Pyr1-apelin-13-mediated beneficial roles in apoptosis and autophagy in CFs associated with decreased AMPK phosphorylated levels and elevated mTOR phosphorylated levels.
CONCLUSIONS In summary, miRNA-122-5p plays an important role in myocardial fibrosis, apoptosis, autophagy and CFs dysfunction. Our data illustrated that miRNA-122-5p exacerbated Ang II-induced profibrotic, proapoptotic and anti-autophagic actions. Pyr1-apelin-13 activation and miRNA-122-5p inhibition appear to show counter regulation against Ang II-mediated loss of autophagy and promotion of apoptosis and oxidative injury in rat CFs by modulating the AMPK-mTOR phosphorylation signaling. Thus, the development of promising strategies to interfere with miRNA-122-5p might be a promising field for the therapeutic approach in myocardial fibrosis and related disorders. This research was supported by the General Program and the National Major Research Plan Training Project of National Natural Science Foundation of China (81770253 and 91849111).
GW31-e0576
Shuang Xu, Hangwei Chen, Huaner Ni, Qiuyan Dai
Department of Cardiology, Shanghai Genaral Hospital, Shanghai Jiao Tong University School of Medicine
OBJECTIVES In atherosclerotic development, nicotine activates inflammation and pyroptosis. Histone deacetylase 6 (HDAC6) is considered to participate in inflammation, however, whether HDAC6 mediates nicotine-induced pyroptosis is largely unknown. This study aims to investigate the role of HDAC6 in nicotine-induced pyroptosis in macrophages.
METHODS For in vivo study, macrophage pyroptosis in plaque was assessed by Tunel/CD68 and Caspase-1/CD68 staining. For in vitro study, pyroptosis and related signaling pathway in RAW264.7 cells were evaluated by western blotting, immunofluorescence, lactic dehydrogenase (LDH) activity, co-immunoprecipitation and chromatin immunoprecipitation.
RESULTS High fat diet and nicotine upregulates macrophage pyroptosis in atherosclerotic lesions. Nicotine promotes pyroptosis in RAW264.7 cells, as evidenced by Caspase-1 cleavege, IL-1β and IL-18 production, elevation of LDH activity and propidium iodide positive rate. In addition, nicotine stimulates the expression of HDAC6, and HDAC6 deficiency by siRNA and Tubastatin-A suppresses nicotine-induced pyroptosis. Moreover, HDAC6-mediated deacetylation of p65 enhances its nuclear translocation and binding to NLRP3 promoter regions. Silencing p65 or NLRP3 resulted in decreased pyroptosis.
CONCLUSIONS HDAC6 inhibition exhibited a protective role against nicotine-induced pyroptosis in macrophages, which is partly mediated by acetylation of p65.
GW31-e0607
Mingyan Shao, Xue Tian, Pengrong Gao, Yong Wang
Beijing University of Chinese Medicine
OBJECTIVES β-Elemene has been widely used as a traditional medicine for its anti-tumor activity against a broad range of cancers. However, the effect of β-elemene on in heart failure mice has yet to be determined. This study aims to investigate the effects of β-elemene on heart failure and its underlying mechanism.
METHODS Left anterior descending (LAD)-induced HF mouse model in vivo and oxygen-glucose deprivation/recovery (OGD/R)-induced H9C2 model in vitro were established. HE staining was used to observe the morphology of heart; using oil red O staining to identify spatial distribution of lipids in mouse cardiac tissue; transcript levels of mitochondrial and peroxisomal FAO genes were assayed by real-time polymerase chain reaction (PCR) and the protein expression was measured by western blot and immunocytochemistry methods.
RESULTS β-Elemene improved heart function by up-regulating cardiac ejection fraction (EF) and fractional shortening (FS) values. Furthermore, β-elemene administration rescued ventricular dilation, lipid accumulation and inflammation infiltration in arginal areas of myocardial infarction. Moreover, β-elemene augmented the mRNA expression of fatty acid oxidation-associated genes, such as peroxisome proliferator-activated receptor β (PPARβ), carnitine palmitoyltransferase 1 (CPT1) and so on. Similar results were obtained in vitro, treatment with β-elemene increased the proteins expressions of PPARβ and CPT1 and suppressed inflammatory markers, such as nuclear factor κB (NF-κB) nuclear translocation, inhibitory κBα (IκBα) degradation, interleukin-6 (IL-6) expression, and pro-inflammatory cytokines (such as TNFα). siRNAs for PPARδ were reversed by β-elemene. Consistently, molecular docking estimate that β-elemene targeted on PPARβ.
CONCLUSIONS This study demonstrated that β-elemene as PPARβ agonist, protected the heart failure mice from lipid-induced inflammatory damage and widened new therapeutic effects for the treatment of heart disease in clinical.
GW31-e0632
Xue Tian, Mingyan Shao, Yong Wang
Beijing University of Chinese Medicine
OBJECTIVES Macrophage polarization plays an essential role in the process of heart injury and repair, and is important for the regulation of inflammation after myocardial infarction (MI). This study aims to clarify whether nano-simvastatin can regulate macrophage polarization to exert anti-inflammatory effect and explore potential mechanism.
METHODS In our study, we constructed left anterior descending (LAD)-induced MI mice models and LPS (100 ng/mL) stimulated Raw264.7 cells for 24 h to induce M1 macrophage activation. In vivo, mice received either ddH2O or nano-simvastatin (1 mg/kg) for 7 days. Echocardiography was used to evaluate cardiac function after MI. HE staining was used to observe the morphology of heart. MPI/MRI molecular imaging dynamically detected the distribution of drugs. In vitro, macrophage polarization pathway-related proteins (such as, CD206, STAT6, Arg-1, IRF4) were detected by Western blot and qPCR. Moreover, flow cytometry were applied in vivo and vitro to detect macrophage surface markers (Ly6Clow, Ly6Chigh/CD206, CD86) to measure the polarization state of macrophages.
RESULTS In vivo, nano-simvastatin is significantly condensed in the MI (area in risk) by MPI/MRI molecular imaging technology. Nano-simvastatin could improve cardiac function by up-regulating cardiac ejection fraction (EF) and fractional shortening (FS) values. The results of HE showed that nano-simvastatin could improve cardiac remodeling and inflammatory cell infiltration. The results of flow cytometry showed that the number of M1 macrophage surface markers were higher than that in the sham-operated mice. Treated by nano-simvastatin, M1 macrophages in the MI (area in risk) were inhibited and M2 macrophages were activated, suggesting that nano-simvastatin regulated cardiac macrophage polarization. In vitro, the results of western blot and qPCR showed that nano-simvastatin could up-regulated the markers of M2 macrophages (CD206, Arg-1), and down-regulated the markers of M1 macrophages (CD86), suggesting that nano-simvastatin could promote the conversion of M1 macrophages to M2 macrophages. Meanwhile, nano-simvastatin could increase the expressions of STAT6 and IRF4. Flow cytometry displayed that the number of M2 macrophages (CD206) in nano-simvastatin group was increased and M1 macrophage (CD86) was decreased compared with model group.
CONCLUSIONS This study identified that nano-simvastatin targeted macrophage polarization to reduce inflammation, alleviate myocardial fibrosis, and improve cardiac remodeling.
GW31-e0647
Yang Sun, Zijian Li
Department of Cardiology and Institute of Vascular Medicine, Peking University Third Hospital; Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Ministry of Health; Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education; Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing 100191, China
OBJECTIVES Pathological cardiac fibrosis is a common pathological feature of many cardiac diseases and an independent risk factor for cardiac morbidity and mortality. Dysfunction of TβRI receptors have been implicated in the pathogenesis of fibrotic diseases. Smad7 acts as scaffold to regulate TβRI degradation in plasma membrane and prevent the downstream of TGF-β, thus targets cardiac fibrosis. However, the mechanism how Smad7 is recruited to modulate receptors degradation is uncovered. Here, we explore whether HIP-55, HPK1-interacting protein of 55 kDa (HIP-55), its amplification confers poor prognosis in individuals with heart failure in our previous works, could regulates cardiac fibrosis by TβRI degradation and the molecular mechanisms underlying HIP-55 governs TβRI.
METHODS HIP-55 knockdown adenovirus and overexpression adenovirus were constructed to transfect primary mice cardiac fibroblasts (MCF). Biochemical experimental including western blot, real-time PCR were performed to test protein and gene expression. To explore degradation, ubiquitination assay and CHX half-time assay were performed. Immunofluorescence, Co-IP and GST pull down were used to dissect protein interaction. Transverse aorta constriction (TAC) were performed on wild type and HIP-55-deficient C57/BL6 mice to simulate TGF-β induced pathological process, control group were executed sham operation. Echocardiography were used to examine the ventricular contractibility and diastolic functions. Collagen fibers were stained with Picric acid-Sirius red to evaluate myocardial fibrosis.
RESULTS As an adaptor protein, HIP-55 conferred to resistant in cardiac fibrosis via recruiting Smad7 and this process is primarily attributable to promote Smad7/TβRI complex formation and accelerates TβRI receptor degradation. Besides that, HIP-55 could stabilize Smad7 by inhibiting ubiquitination-dependent degradation. These two regulation of HIP-55 impairs TβRI stability and dampens activated TGF-β signaling. These inhibitory effects were dependent on Smad7, as Smad7 ablation abolished them. The against cardiac fibrosis effects of HIP-55 were further confirmed in TAC induced cardiac fibrosis mice model. Cardiac fibrosis was highly exacerbated, cardiac functions whether in ventricular contractibility or diastolic function of HIP-55-decifient mice were worse than wild type mice including lower LVEF and LVFS and higher E/E'. And fibrosis area of HIP-55-decifient mice was remarkable more than wild type mice.
CONCLUSIONS The results presented here suggest that HIP-55 could recruit Smad7 to plasma membrane to accelerate TβRI receptors ubiquitin-dependent degradation. On the other hand, HIP-55 could also form complex with Smad7 to impede Axin/Smad7/Arkadia complex formation and consequently modulation of Smad7 protein stability. By harnessing the mechanism of HIP-55 regulating TβRI receptors could develop putative prophylactic and therapeutic strategies against cardiac fibrosis.
GW31-e0654
Guohua Ni1, Meifang Chen2
1Sohome Health Management Center, Sichuan Provincial People’s Hospital
2Xiangya Hospital, Central South University
OBJECTIVES Asymmetric dimethylarginine (ADMA) is a naturally occurring substance, it inhibits the production of nitric oxide (NO) synthesis in vivo. Profilin-1, a small actin-binding protein, has been documented to be involved in endothelial injury and the proliferation of vascular smooth muscle cells (VSMCs) in hypertension. To investigate the relationship between ADMA and profilin-1 in hypertensive individuals and in cultured VSMCs.
METHODS Forty healthy subjects and 42 matched essential hypertensive patients took part in this study. Rat aortic smooth muscle cells (RASMCs) were treated with different concentrations of ADMA as indicated for 24 h or 30 μM ADMA for different periods of time. RASMCs were transfected with profilin-1 shRNA to interrupt protein expression of profilin-1. The expression of profilin-1 in RASMCs was tested via real time-PCR and western blot analysis. RASMCs were treated with ADMA after pretreated with AG490 (5×10-5 M) or rapamycin (10-8 M). The level and expression of profilin-1 in RASMCs were tested by real time-PCR and western blot analysis. Cell proliferation was measured via flow cytometry analysis and 3-(4,5-dim-ethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.
RESULTS Compared with healthy subjects, the levels of ADMA, profilin-1, tumor necrosis factor (TNF-α), vWF and interleukin-8 (IL-8) were markedly elevated, and the levels of nitric oxide (NO) were significantly decreased in hypertensive individuals (P<0.05). In vitro, ADMA induced the expression of profilin-1 in concentration-dependent and time-dependent manners in RASMCs (P<0.05). ADMA-induced proliferation of RASMCs was significantly inhibited by knockdown of profilin-1 by short hairpin RNA. Furthermore, ADMA-mediated proliferation of RASMCs and expression of profilin-1 were inhibited by blockade of JAK2/STAT3 pathway.
CONCLUSIONS Profilin-1 may be involved in ADMA-mediated vascular lesion in hypertension.
GW31-e0681
Xiaoxiao Zhao1, Haneul Cho2, Sora Lee2, Weon Kim2
1Taizhou Hospital of Zhejiang Province, Taizhou, China
2Kyung Hee University Medical Center Hospital Center Cardiological Lab, Souel, Korea
OBJECTIVES Doxorubicin (DOX) administration decreases cardiac soluble guanylate cyclase (sGC) activity by reactive oxygen species (ROS) storm. We hypothesized direct sGC stimulation ameliorated doxorubicin-cardiotoxicity. The present study investigated the mechanism and therapeutic effect of sGC activation.
METHODS An activator of oxidized and deactivated sGC, BAY-602770, was used to direct stimulate sGC. H9c2 cardiomyoblasts were treated with DOX (0.5–10 μM), with or without pre-treated 10 μM BAY 60-2770. SD rats were orally administered with BAY60-2770 1 hour prior to every DOX treatment. Echocardiography and hemodynamic values were then analyzed. Proteins expression levels were examined by western blot analysis.
RESULTS BAY60-2770 improved cell viability and ROS in H9c2 cells expose to DOX, which was mediated by PKG1 activation. DOX-induced caspase-3 activation decreased after pretreatment with BAY60-2770 in vivo and in vitro. Mitochondrial ROS and TMRE fluorescence were attenuated by BAY60-2770 associated with high level mitochondrial ferritin (MtFt) expression. We constructed MtFt knock down (MtFt-KD) cells by using siRNA, subsequently did Cyto-ID autophagy detection. Autophagosome was decreased by MtFt-KD, however, BAY60-2770 offset disparate autophagy level between MtFt-KD cells and non-MtFt-KD cells exposed to DOX. As animal experiments, echocardiography showed that BAY60-2770 significantly improved DOX-induced myocardial dysfunction.
CONCLUSIONS PKG1 activation participated in myocardial protection against DOX cardiotoxicity, which associated with MtFt upregulation. MtFt could be one of the antioxidant mechanisms to prevent sGC and PKG1 from oxidative inactivation.
GW31-e0694
Liuying Zheng, Hongliang Cong
Tianjin Chest Hospital
OBJECTIVES In 2016, the European Heart Rhythm Association and four other associations established an expert consensus to define, characterise, and classify atrial cardiomyopathy into 4 subgroups based on their histopathological features. The predominant pathological feature of class I and III atrial cardiomyopathy is the hypertrophy of atrial cardiomyocytes. Angiotensin II (AngII) is a major effector peptide of the renin–angiotensin system (RAS) and it commonly stimulates hypertrophy. Emerging evidence suggests that epigenetic mechanisms are involved in RAS-induced cardiac remodelling. Epigenetic regulation such as histone (H3 and H4) lysine acetylation (H3Kac and H4Kac) plays a central role in gene transcription. Histone deacetylation by histone deacetylases (HDACs) promotes chromatin condensation, which causes transcriptional repression. We studied the epigenetic mechanisms by which AngII promotes atrial cardiomyopathy progression. We hypothesise that AngII induces the nuclear export of HDAC-4 and -5 and leads to the derepression of hypertrophy-related genes.
METHODS Atrial tissues were obtained from patients who underwent surgeries. Atrial cardiomyocytes were isolated from the hearts of neonatal rats. The effects and mechanisms of AngII-induced atrial cardiomyocyte hypertrophy were evaluated by enzyme-linked immunosorbent assay (ELISA), immunofluorescence, western blotting, RNA-seq, and chromatin immunoprecipitation (ChIP).
RESULTS Compared with that of sinus rhythm (SR) control individuals, the myocardium of patients with atrial fibrillation (AF) exhibited increased levels of AngII, chromatin-bound myocyte enhancer factor 2 (MEF2), H4ac, and H3K27ac; upregulation of hypertrophy-related genes; and decreased levels of HDAC-4 and -5 bound to the promoters of hypertrophy-related genes. Furthermore, incubation of atrial cardiomyoctytes with AngII increased their cross-sectional area and stimulated the expression of hypertrophy-related genes. AngII also promoted the phosphorylation of HDAC4 and HDAC5 and induced their nuclear export. RNA-seq analyses revealed that AngII significantly up-regulated genes associated with cardiac hypertrophy. ChIP revealed that AngII increased the levels of chromatin-bound MEF-2, H4ac, and H3K27ac and decreased HDAC-4 and -5 enrichment on the promoters of hypertrophy-related genes. All these AngII-induced pro-hypertrophic effects could be partially reverted by losartan (AngII receptor blocker).
CONCLUSIONS Patients with AF manifest an increased susceptibility to hypertrophy and exhibit epigenetic characteristics that are permissive for the transcription of hypertrophy-related genes. MEF2 may act as platform to respond to positive or negative transcriptional signals by exchanging HATs and class II HDACs. AngII induces histone acetylation via the cytoplasmic-nuclear shuttling of HDACs, which is a novel mechanism of atrial hypertrophy regulation and might provide a promising therapeutic strategy for atrial cardiomyopathy.
GW31-e0705
Yangxi Hu1,2, Rongfang Zhu3, Yongwen Qin1, Xianxian Zhao1, Chun Liang2, Qing Jing1,3
1Department of Cardiology, Shanghai Changhai Hospital, Naval Medical University
2Department of Cardiology, Shanghai Changzheng Hospital, Naval Medical University
3Shanghai Institutes for Biological Sciences, University of the Chinese Academy of Sciences
OBJECTIVES Angiogenesis is a key process for establishing functional vasculature during embryogenesis and involves different signaling mechanisms. The RNA binding protein Zfp36l1 was reported to be involved in various diseases in different species, including cardiovascular diseases. However, whether Zfp36l1b, one of the 2 paralogs of Zfp36l1 in zebrafish, works like mammalian Zfp36l1, and if their molecular mechanisms are different remain unclear. Here, we show that Zfp36l1b plays a crucial protective role in angiogenesis of zebrafish embryos.
METHODS We used transparent transgenic and wild-type zebrafish larvae to dynamically investigate the early stage of angiogenesis with confocal in vivo, after the knockdown of Zfp36l1b by morpholinos (MOs). In situ hybridization and fluorescence-activated cell sorting were performed to detect Zfp36l1b expression. mRNA rescue and CRISPR/Cas9 knockdown, and luciferase reporter experiments were performed to further explore the role of Zfp36l1b in angiogenesis.
RESULTS We found that knockdown of Zfp36l1b led to defected angiogenesis in intersomitic vessels and sub-intestinal veins (SIVs), which could be rescued by the supplement of Zfp36l1b mRNA. Moreover, knockdown of Zfp36l1b suppressed Notch1b expression, while knockdown of Notch1b resulted in partly relief of angiogenesis defects induced by Zfp36l1b down-regulation. Besides, Zfp36l1b knockdown alleviated the excessive branch of SIVs caused by Vegfa over-expression.
CONCLUSIONS Our results show that Zfp36l1b is responsible for establishing normal vessel circuits through affecting the extension of endothelial tip cells filopodia and the proliferation of endothelial cells partly through Notch1b/Fll4 suppression and synergistic function with Vegfa.
GW31-e0706
Yangxi Hu1,2, Changzhen Ren2, Rongfang Zhu3, Yanda Zhang2, Zhiqing He2, Yongwen Qin1, Xianxian Zhao1, Chun Liang2, Qing Jing1,3
1Department of Cardiology, Shanghai Changhai Hospital, Naval Medical University
2Department of Cardiology, Shanghai Changzheng Hospital, Naval Medical University
3Shanghai Institutes for Biological Sciences, University of the Chinese Academy of Sciences
OBJECTIVES ApoEb is a characterized zebrafish gene homologous to mammalian ApoE. ApoE (Apolipoprotein E) is recognized as a ligand for the LDL receptor (LDLR), whose deficiency would largely lead to atherosclerosis and other cardiovascular diseases. Here, we attempted to knock out the zebrafish ApoEb gene using the CRISPR/Cas9 system and high-fat diet (HFD) feeding to construct a zebrafish model with a disorder of lipid metabolism.
METHODS The establishment of the model was confirmed by Oil Red O (ORO) staining and lipid measurement. We also detected whether ApoEb deficiency activated the sterol-regulatory element binding protein (SREBP) pathways or induced the malfunction of LDLR. At last, using dimethyl sulfoxide (DMSO) as a negative control and Simvastatin as a positive control, the model was treated with Xuezhikang (XZK), an extract derived from red yeast rice, which is commonly employed as a traditional Chinese medicine for treating coronary heart disease, decreasing blood lipids and preventing other cardiovascular events both within China and globally. ORO staining, lipid measurement of tissue homogenate, and fluorescence microscopy photography were used to obverse its effect on zebrafish larvae.
RESULTS The results demonstrated that in the mutant, significant lipid deposition in the blood vessels occurred, and lipid measurement of larvae’s whole-body homogenate and adult plasma showed significant increase in total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) levels. Interestingly, these lipid metabolism disorders could be apparently relieved by Xuezhikang treatment, and the recruitment of neutrophils in the blood vessels was significantly inhibited.
CONCLUSIONS In conclusion, a lipid metabolism disorder model has been successfully constructed in zebrafish through ApoEb knockout. Using this novel model, we found that the preventive effect on atherosclerosis of XZK may be caused by lowering blood lipids and inhibiting the recruitment of neutrophils in blood vessels.
GW31-e0707
Yangxi Hu1,2, Xia Zhu1,3, Shaohua Dong1, Jianliang Zhang1, Xianxian Zhao1, Chun Liang2, Lujun Zhang1
1Department of Cardiology, Shanghai Changhai Hospital, Naval Medical University
2Department of Cardiology, Shanghai Changzheng Hospital, Naval Medical University
3Department of Cardiology, Urumqi Friendship Hospital
OBJECTIVES Hyperhomocysteinemia (HHcy) has emerged as an independent risk factor for vascular diseases. Previous observations demonstrated that homocysteine (Hcy) contributes to the arrest of endothelial cell (EC) growth, but the precise underlying mechanism is unclear. The present study focused on the role of miR-21 in Hcy-induced human umbilical vein ECs (HUVECs) and explored its underlying mechanisms.
METHODS HUVECs were treated with 0 mM, 3 mM or 5 mM Hcy, and then apoptosis and cell cycle were assessed through flow cytometry. Rt-qPCR was used to detect the expression levels of 15 miRNAs and 15 target genes of miR-21. Luciferase reporter assays and Western blot were used to confirm the down-regulation of miR-21 on cell division cycle 25A (CDC25A) expression. Flow cytometry revealed how lentivirus induced CDC25A overexpression and miR-21 mimics affected cell cycle progression.
RESULTS Treating HUVECs with Hcy caused cell cycle arrest and inhibited cell proliferation. Hcy treatment increased miR-21 levels, which repressed the expression of CDC25A. Overexpression of miR-21 downregulated CDC25A expression, whereas miR-21 inhibition up-regulated its expression. Luciferase assays showed that miR-21 regulated CDC25A level by binding to the 3'-UTR. MiR-21 overexpression induced cell cycle arrest and cell proliferation in Hcy-treated HUVECs. However, overexpressing CDC25A could promote cell proliferation and cell cycle, overcoming the effects of Hcy-induced miR-21 upregulation.
CONCLUSIONS This study demonstrated that miR-21 plays an important role in modulating cell cycle progression and cell proliferation by targeting CDC25A in an Hcy-induced cell and suggests new insights into the prevention of Hcy-associated cell dysfunction.
GW31-e0708
Shuai Shao, Jiguang Wang
Shanghai Key Laboratory of Hypertension, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
OBJECTIVES Hypertension is a leading risk factor for morbidity and mortality of cardiovascular disease. Genome-wide association studies have identified that natriuretic peptide receptor-C (NPR-C) variants are associated with elevation of blood pressure. However, whether NPR-C regulates blood pressure through sodium homeostasis is unknown. In this study, we tested the hypothesis that NPR-C signaling regulates the sodium retention-related hypertension.
METHODS AngII infusion hypertension mouse model was used in male and female wild type (WT) and NPR-C knockout (KO) mice. Systolic blood pressure was measured by tail cuff method. Acute saline expansion was performed in mice after AngII treatment. Renal sodium transporters expression and associated signaling pathway were evaluated by Western Blot. Renal Na+ clearance was measured by NaCl cotransporter (NCC) blocker hydrochlorothiazide. Different dietary Na+ intake include 4% high Na+ and 0.02% low Na+ were used to determine the role of NPR-C in the regulation of NCC activity. The effect of NPR-C on blood pressure was also evaluated in renal tubule-specific deletion of NPR-C mice were also infused with AngII. CoroNa Green, a sodium ion indicator, and flow cytometry was used to measure sodium uptake in distal convoluted tubule cells. In order to study the mechanism of NPR-C regulating water-salt metabolism, the changes of WNK4/SPAK/NCC expression in distal convoluted tubule cells were detected after giving the inhibitors of NPR-C upstream include natriuretic peptide system and downstream signal such as inhibitory G protein (Gi), phospholipase C (PLC), protein kinase C (PKC) respectively.
RESULTS (1) AngII infusion increased both blood pressure and sodium retention throughout two weeks infusion period both in male and female wild type mice. NPR-C gene knockout alleviates AngII-induced hypertension by inhibiting NCC activity and facilitating urinary sodium excretion. NPR-C deficiency led to natriuresis in response to acute saline expansion after treatment of AngII. Interestingly, AngII increased both total and phosphorylation of NCC abundance involving in activation of With-No-lysine Kinase 4 (WNK4)/Ste20 related proline/alanine rich kinase (SPAK) which was blunted by NPR-C deletion. NCC inhibitor, hydrochlorothiazide, failed to induced natriuresis in NPR-C knockout mice. Moreover, low salt and high salt diets-induced changes of total and phosphorylation of NCC expression were normalized by NPR-C deletion. Importantly, tubule-specific deletion of NPR-C also attenuated AngII-induced elevated blood pressure, total and phosphorylation of NCC expression. In primary cultured distal convoluted tubule cells, AngII upregulated NCC-mediated sodium uptake was suppressed by NPR-C knockdown. (2) Mechanistically, in distal convoluted tubule cells, AngII dose and time-dependently upregulated WNK4, p-SPAK, NCC and p-NCC expression. NPR-C signaling but not NPR-B signaling pathway mediated NCC activation. NPR-C regulates AngII-induced activation of WNK4/SPAK/NCC proteins expression via the Gi/PLC/PKC signal pathway.
CONCLUSIONS These results demonstrated a novel mechanism through which NPR-C deficiency decreases AngII-induced elevation of blood pressure by suppressing NCC-dependent sodium reabsorption in the kidney. These experiments further identify NPR-C as a potential strategy for the treatment of sodium retention-related hypertension.
GW31-e0723
Zuowei Pei, Fang Wang
Department of Cardiology, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences
OBJECTIVES Hyperlipemia is a risk factor for cardiac damage and cardiovascular disease. Several studies have presented that thymoquinone (TQ) can protect cardiac damage. The aim of the study was to investigate the possible protective effects of TQ reduced hyperlipemia-induced cardiac damage in low-density lipoprotein receptor deficient (LDL-R- /- ) mice.
METHODS Eight-week-old male LDL-R-/- mice were randomly divided into the following three groups: the control group with a normal diet (ND group), the high fat diet (HFD) group, and the HFD mixed with TQ (HFD+TQ) group. All groups were used the different diets for 8 weeks. Blood samples were collected in serum tubes, and stored at 2280 °C until use. Cardiac tissues were stored in 10% formalin and then embedded in paraffin for histological evaluation. The remainder of the cardiac tissues was snap-frozen in liquid nitrogen for mRNA preparation or immunoblotting.
RESULTS The levels of metabolism-related factors, low-density lipoprotein-cholesterol, total cholesterol and high-sensitivity C-reactive protein, were decreased in the HFD+TQ group compared with that in the HFD group. Periodic acid-Schiff staining demonstrated that lipid deposition was lower in the HFD+TQ group than that in the HFD group. The expression of pyroptosis indicators (NLRP3, IL-1β, IL-18 and caspase-1), pro-inflammation factors (IL-6 and TNF-α), and macrophage markers (CD 68) was significantly downregulated in the HFD+TQ group compared with that in the HFD group.
CONCLUSIONS Our results indicate that TQ can serve as a potential therapeutic agent for hyperlipemia-induced cardiac damage.
GW31-e0726
Wenbin Fu1,2,3,4, Chunyu Zeng1,2,3,4
1Department of Cardiology, Daping Hospital, The Third Military Medical University, Chongqing 400042, China
2Department of Cardiology, Daping Hospital, Army Medical University, Chongqing 400042, China
3Chongqing Institute of Cardiology, Chongqing 400042, China
4Cardiovascular Research Center, Chongqing College, University of Chinese Academy of Sciences, Chongqing 400042, China
OBJECTIVES The mammalian cardiomyocyte is capable of regeneration for a brief window of time after birth, but the regenerative capacity is lost in adults, which is responsible for irreversible heart failure after myocardial infarction (MI). The molecular mechanisms controlling endogenous cardiac regeneration remain largely elusive. Multifaceted functions of long non-coding RNAs (lncRNAs) have been indicated in governing regulatory network of cardiac development and diseases, therefore we screened and verified novel lncRNAs in regulating cardiomyocyte proliferation and cardiac regeneration.
METHODS Using a neonatal cardiac regeneration model of apex resection (AR), we identified a lncRNA, named DRR (DNA repair regulator), that possessed a negative effect in regulating cardiomyocyte proliferation. Next, we generated lncRNA-DRR knockout mice through CRISPR system and performed mouse models of AR and MI. To explore the function and mechanisms of lncRNA-DRR in cardiomyocyte proliferation, various analyses were carried out including echocardiographic evaluation, histology immunofluorescence staining, quantitative real-time polymerase chain reaction, fluorescence in situ hybridization (FISH), western blot, immunoprecipitation and bioinformatics analysis.
RESULTS LncRNA-DRR was significantly reduced after AR in neonates, and overexpression of lncRNA-DRR decreased while silencing of lncRNA-DRR increased the proliferation of cultured cardiomyocytes. In addition, lncRNA-DRR deletion unaffected normal heart development but was sufficient to prolong postnatal window of regeneration capacity after AR injury. In adults, lncRNA-DRR deletion improved cardiac function and reduced infarct size in post-MI hearts, which was associated with significant improvement of cardiomyocyte proliferation mediated cardiac regeneration. Next, gene enrichment analysis revealed that cell cycle process and cellular response to DNA damage were up-regulated in hearts of lncRNA-DRR-/- mice. Further computational analysis and immunoprecipitation showed SFPQ, a DNA repair related molecule, was a target of lncRNA-DRR, and their binding site overlaps with the coiled coil domain of SFPQ which mediates interaction with NONO. LncRNA-DRR deletion could promote the binding of SFPQ-NONO heteromer, decrease DNA damage and activate cardiomyocyte cell-cycle re-entry.
CONCLUSIONS Collectively, our results identified that lncRNA-DRR was a negative regulator of cardiomyocyte proliferation, and loss of lncRNA-DRR promoted heart regeneration through stabilizing SFPQ-NONO heteromer induced DNA repair. LncRNA-DRR might be a novel target for stimulating cardiac regeneration and further MI treatment.
GW31-e0754
Yingying Zhang
Tianjin Chest Hospital
OBJECTIVES In previous studies, we found that decreased miR-223 level predicts high on-treatment platelet reactivity (HTPR) in patients with troponin-negative non-ST elevation ACS. But, some studies have shown that the level of miR-223 in the plasma are reduced following inhibition of platelet function. Accordingly, it is unclear decreased plasma or platelet miR-223 levels appear a marker of low or high responsiveness to DAPT. Furthermore, the relationship between the circulating miR-223 level and future clinical outcomes in ACS patients is obscure. Given the importance of the miRNA in platelet reactivity. The present study investigated whether the plasma miR-223 level can predict future MACE accure in ACS patients after PCI.
METHODS One hundred and eighty-eight consecutive NSTE-ACS patients undergoing PCI and on DATP were enrolled in this study and they were dichotomized according to the medians of their circulating miR-223 levels (group 1: miR-223 level >0.993, n=94; group 2: miR-223 level <0.993, n=94). The plasma miR-223 level was quantified by real-time PCR, and the platelet reactivity was determined by platelet reactivity index (PRI), measured by vasodilator-stimulated phosphoprotein (VASP) phosphorylation flow cytometry at least 2 hours after 600 mg clopidogrel plus aspirin treatment.
RESULTS Compared with group1, PRI level was significantly elevated in group 2 (45.0±19.3 vs. 54.6±18.5, P<0.01**). In addition, circulating miR-223 level was inversely correlated with PRI (Spearman r=−0.400, P<0.01). Cox regression analysis revealed that among factors that potentially influence Major Adverse Cardiac Events (MACE), decreased circulating miR-223 level (HR: 0.137, 95% CI: 0.040–0.462, P<0.05*) and PRI (HR: 1.038, 95% CI: 1.010–1.067, P<0.01**) were independent predictors for the presence of MACE during 24 months follow-up. ROC curve analysis also positive the ability of miR-223 (AUC: 0.804, 95% CI: 0.702–0.907, P<0.01) and PRI (AUC: 0.775, 95% CI: 0.680–0.870, P<0.01) in predicting MACE.
CONCLUSIONS Our data suggest that circulating miR-223 level may serve as novel biomarker for assessment of clopidogrel responsiveness and predicting the clinical ischemic outcomes for ACS patients with DAPT after PCI during 24 months follow up.
GW31-e0755
Siang Wei1, Zhiwen Ding1, Ran Xu1, Yuyao Ji1, Yan Feng2, Yunzeng Zou1
1Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University
2Shanxi Agricultural University
OBJECTIVES To study the relationship between hexosamine metabolic pathway and myocardial hypertrophy, and to provide new ideas and targets for the treatment of hypertrophic cardiomyopathy.
METHODS After 24 hours of primary culture of neonatal SD rat cardiomyopathy for 1–3 days, they were randomly divided into five groups, and 0.1% angiotensin II stimulated cardiomyopathy to construct a hypertrophy model. The acting time was 0, 6, 12, 24, and 48 hours, respectively. The mRNA expression levels of ANP, BNP, and GFPT2, the crucial factors of hexosamine metabolism and cardiac hypertrophy, were detected after 48 hours of culture. Then the physical and chemical properties, amino acid composition, GO, and KEGG of the GFPT2 gene were analyzed by bioinformatics software.
RESULTS ANP mRNA and BNP mRNA increased significantly with the prolongation of angiotensin II stimulation time, and reached the highest at 48 h. GFPT2 mRNA expression increased at 6, 12, 24, and 48 h after angiotensin II stimulation, and reached the highest at 48 h. Human GFPT2 protein contains 682 amino acids with a single-chain structure, also mainly including α and β structures. The isoelectric point/molecular weight was 7.03/76930.60. There are eight positive selection sites for GFPT2 amino acid codons. In addition, the phosphorylation sites, acetylation sites, and ubiquitination sites related to diseases are 10, 2, and 5. GO enrichment analysis of GFPT2 is mainly involved in the biological processes of GFPT enzyme activity, protein binding, carbohydrate derivative binding. KEGG analysis showed that the gene was fundamentally involved in amino sugar and nucleotide sugar metabolism, alanine, aspartic acid, and glutamic acid metabolism, and insulin resistance. According to the scores of edges’ LLS, the top 10 genes were NNMT, ADAM12, BGN, CoL5A1, MMP2, CRISOLD2, VCAN, MXKA5, PRRX1, CoL5A2.
CONCLUSIONS high expression of hexosamine metabolism is induced by myocardial hypertrophy, which is aggravated with the prolongation of angiotensin II stimulation time, and the metabolic disorder may be the main cause of hypertrophic cardiomyopathy.
GW31-e0756
Jiatong Wu1, Zhuo Ao2, Quanmei Sun2, Maojing Shi1, Cheng Cheng1, Dong Han2, Yuansheng Liu1
1Emergency Department, Peking University People’s Hospital
2National Center for Nanoscience and Technology
OBJECTIVES To investigate the effect of micro-scale stretch stimulus on hypertension-associated hypertrophied ventricular myocytes.
METHODS Separating respectively ventricular myocytes from spontaneously hypertensive rats (SHRs) and homologous normotensive rats (Wistar-Kyoto, WKY) of 25-week-old, and dying intracellular calcium with fluo-4/AM, then, applying micro-scale stretch stimulus on single ventricular myocytes and observing the change of intracellular calcium average fluorescent intensity synchronously via an united imaging system of atomic force microscope and laser scanning confocal microscope (Bio-AC). On the other hand, adding streptomycin into ventricular myocytes loaded fluo-4/AM, obtaining the change of intracellular calcium average fluorescent intensity with laser scanning confocal microscope.
RESULTS After the same micro-scale stretch stimulus, both intracellular calcium average fluorescent intensity of ventricular myocytes from SHRs and WKY rats was increased significantly than before (3527.29±217.18 vs. 4891.79±648.65, 3447.02±151.58 vs. 4067.70±247.95, P<0.01). Whether before, during or after the application of micro-scale stretch stimulus, the intracellular calcium average fluorescent intensity of ventricular myocytes separated from SHRs was higher than that from WKY rats (3527.29±217.18 vs. 3447.02±151.58, 4322.20±265.01 vs. 3391.80±139.43, 4891.79±648.65 vs. 4067.70±247.95, P<0.01). Meanwhile, after stretch stimulus, comparing with WKY rats, the intracellular calcium average fluorescent intensity of ventricular myocytes from SHRs had a bigger elevation (39.49±4.90% vs. 18.19±8.21%, P<0.01). Besides, the response of ventricular myocytes from SHRs to stretch stimulus was more rapid than that from WKY rats. In the intervention of streptomycin, both of intracellular calcium average fluorescent intensity of ventricular myocytes from SHRs and WKY rats had significantly decreased (2512.81±320.81 vs. 1976.16±194.40, 2176.90±244.01 vs. 1910.92±38.01, P<0.01), however, the decreased degree of the former was notable than the latter (19.78±14.80% vs. 11.30±8.31%, P<0.01).
CONCLUSIONS Hypertrophied ventricular myocytes are more sensitive to stretch stimulus, and have a higher cellular calcium concentration more easily in the presence of mechanical stimulation, thus, the property can promote the occurrence of cardiac arrhythmias. Besides, the stretch-activated channels on hypertrophied ventricular myocytes have been increased or more powerful, and this change possibly takes an important part in the occurrence and maintenance of cardiac arrhythmias.
GW31-e0771
Xuewei Xia, Chunyu Zeng
Army Medical Center of PLA
OBJECTIVES Abnormal migration of vascular smooth muscle cells (VSMCs) serves an important role in hypertension, atherosclerosis and restenosis following angioplasty, which is regulated numerous hormonal and humoral factors, including neuropeptide Y (NPY) and dopamine. Dopamine and NPY are both sympathetic neurotransmitters, and a previous study reported that NPY increased VSMC proliferation, while dopamine receptor inhibited it. Therefore, the authors wondered whether or not there is an inhibitory effect of dopamine receptor on NPY-mediated VSMC migration.
METHODS The migration of VSMC was detected by transwell migration assay and wound healing assay.
RESULTS The present study demonstrated that stimulation with NPY dose-dependence (10-10-10-7 M, 24 h) increased VSMC migration, the stimulatory effect of NPY was via the Y1 receptor. This is because, in the presence of the Y1 receptor antagonist, BIBP3226 (10-7 M), the stimulatory effect of NPY on VSMC migration was blocked. Activation of the D3 receptor by PD128907 dose-dependence (10-11-10-8 M) reduced the stimulatory effect of NPY on VSMC migration. The effect of PD128907 was via the D3 receptor, because the inhibitory effect of PD128907 on NPY-mediated migration was blocked by the D3 receptor antagonist, U99194. The authors’ further study suggested that the inhibitory effect of the D3 receptor was via the PKA signaling pathway, in the presence of the PKA inhibitor, 14–22 (10-6 M), the inhibitory effect of PD128907 on VSMC migration was blocked. Moreover, the inhibitory effect of PD128907 was imitated by PKA activator, Sp-cAMP [S], in the presence of Sp-cAMP [S], the NPY-mediated stimulatory effect on VSMC migration was abolished.
CONCLUSIONS The present study indicated that activation of the D3 receptor inhibits NPY Y1-mediated migration on VSMCs, PKA is involved in the signaling pathway.
GW31-e0772
Jianjie Dong, Chen Wang, Zuyi Yuan
Department of Cardiology, First Affiliated Hospital, Xi’an Jiaotong University, Xi’an 710061, China
OBJECTIVES Proprotein convertase subtilisin/kexin type 9 (PCSK9) detriments cholesterol homeostasis by targeting hepatic low-density lipoprotein receptor (LDLR) for lysosomal degradation. Clinically, PCSK9 inhibitors effectively reduce LDL cholesterol (LDL-C) and the incidence of cardiovascular events. As microRNA (miR) are understood to be integral regulators of cholesterol homeostasis, we investigate the involvement of miR-483 in regulating LDL-C metabolism.
METHODS In silico methods and in vitro experiments were used to explore miR-483-5p inhibits PCSK9 production with ensuing increase in LDLR expression and LDL-C uptake in hepatocytes. AAV8-mediated hepatic overexpression of miR-483 together with various forms of PCSK9 were used to decipher the cholesterol-lowering effect of miR-483 in hypercholesterolemic mouse models.
RESULTS In HepG2 cells, miR-483-5p targets the PCSK9 3′UTR, which suppressed PCSK9 expression, increases LDLR level, and promotes LDL-C uptake. In hypercholesterolemic mouse models, hepatic miR-483 overexpression increased LDLR expression via PCSK9 targeting. Consequently, serum cholesterol and LDL-C levels drastically decreased. Mechanistically, the cholesterol-lowering effect of miR-483-5p was seen in mice receiving AAV8 PCSK9-3′UTR (WT), but was absent in mice administered AAV8 PCSK9-3CSK9-3′UTR (UTR (ΔBS) in which the miR-483-5p targeting site was deleted nor in LDLR knockout mice.
CONCLUSIONS MiR-483-5p targeting PCSK9 led to increased LDLR expression and LDL uptake in hepatocytes. Exogenously administered miR-483 could drastically ameliorate high-fat diet or PCSK9-induced hypercholesterolemia in mouse models.
GW31-e0773
Jiao Zhang, Chen Wang, Zuyi Yuan
Department of Cardiology, First Affiliated Hospital of Xi’an Jiaotong University
OBJECTIVES Angiotensin-converting enzyme 2 (ACE2) converts angiotensin II (Ang-II), the most potent vasoconstrictor, to Ang 1-7 and is also a membrane protein that enables COVID-19 infectivity. AMP-activated protein kinase (AMPK) phosphorylation of ACE2 enhances ACE2 stability and this mode of post-translational modification (PTM) of ACE2 in vascular endothelial cells (ECs) is causative of a pulmonary hypertension (PH)-protective phenotype. The oncoprotein murine double minute 2 (MDM2) is an E3 ligase that ubiquitinates its substrates to cause their degradation. In this study, we investigate whether MDM2 is involved in the PTM of ACE2 via its ubiquitination of ACE2, and whether an AMPK and MDM2 crosstalk regulates the pathogenesis of PH.
METHODS Bioinformatic analyses were used to explore E3 ligase that ubiquitinates ACE2. Cultured ECs, mouse models, and specimens from patients with idiopathic pulmonary arterial hypertension (IPAH) were used to investigate the crosstalk between AMPK and MDM2 in regulating ACE2 phosphorylation and ubiquitination in the context of PH.
RESULTS Levels of MDM2 were increased and those of ACE2 decreased in lung tissues and lung ECs isolated from IPAH patients and rodent models of experimental PH. MDM2 inhibition by JNJ-165 reverted the SU5416/Hypoxia-induced PH in C57BL/6 mice. ACE2 S680L (dephosphorylation at S680) mice showed PH susceptibility and ectopic expression of ACE2 S680L/K788R (dephosphorylation at S680; deubiquitination at K788) reduced experimental PH. Moreover, ACE2 K788R overexpression in mice with EC-specific AMPKα2 knockout mitigated PH.
CONCLUSIONS Maladapted PTM (phosphorylation and ubiquitination) of ACE2 at Ser-680 and Lys-788 is involved in the pathogenesis of PAH and experimental PH. Thus, a combined intervention of AMPK and MDM2 in the pulmonary endothelium might be therapeutically implicated in PH treatment.
GW31-e0777
Xin Su1,2, Daoquan Peng2
1Xiamen Cardiovascular Hospital of Xiamen University
2Second Xiangya Hospital of Central South University
OBJECTIVES The hallmark of obesity is excessive accumulation of triglyceride (TG) in adipose tissue. Apolipoprotein A5 (ApoA5) has been shown to influence the prevalence and pathogenesis of obesity. However, the underlying mechanisms remain to be clarified.
METHODS Human adipose-derived mesenchymal stem cells (AMSCs) were treated with 600 ng/mL human recombinant ApoA5 protein. The effect of ApoA5 on intracellular TG content and adipogenic related factors expression were determined. Furthermore, the effect of ApoA5 on CIDE-C expression was also detected.
RESULTS During the process of adipogenesis, ApoA5 treatment reduced the intracellular accumulation of lipid droplets and the TG levels; meanwhile, ApoA5 down-regulated the expression levels of adipogenic related factors, including CCAAT enhancer binding proteins α/β (C/EBPα/β), fatty acid synthetase (FAS), and fatty acid-binding protein 4 (FABP4). Furthermore, the suppression of adipogenesis by ApoA5 was mediated through the inhibition of CIDE-C expression, an important factor which promotes the process of adipogenesis. However, over-expressing intracellular CIDE-C could lead to the loss-of-function of ApoA5 in inhibiting AMSCs adipogenesis.
CONCLUSIONS In conclusion, ApoA5 inhibits the adipogenic process of AMSCs through, at least partly, down-regulating CIDE-C expression. The present study provides novel mechanisms whereby ApoA5 prevents obesity via AMSCs in humans.
GW31-e0778
Xin Su1,2, Daoquan Peng2
1Xiamen Cardiovascular Hospital of Xiamen University
2Second Xiangya Hospital of Central South University
OBJECTIVES Obesity is associated with a series of health problems which are always grouped together as metabolic syndromes. The hallmark of obesity is excessive accumulation of triglyceride (TG) in adipose tissue. Apolipoprotein A1 (apoA1) is a recently described protein that has been shown to influence obesity. However, the mechanism involved remains to be fully elucidated. The aim of this study was to examine the anti-obesity effect and the mechanism of apoA1 during the adipogenesis differentiation of human adipose-derived mesenchymal stem cells (AMSCs).
METHODS Human adipose-derived mesenchymal stem cells (AMSCs) were treated with 15 μg/mL wide-type human recombinant ApoA1 protein and 15 μg/mL loss-of-function mutation human recombinant ApoA1 protein. The effect of ApoA1 on intracellular TG content and adipogenic related factors expression were determined. Furthermore, the effect of ApoA5 on intracellular expression levels of sortilin was also detected.
RESULTS Both two type of apoA1 could significantly reduce the accumulation of intracellular lipid droplets, the concentration of TG and inhibited AMSCs differentiation by down-regulating the gene and protein expression of adipogenesis differentiation-related factors, including CCAAT enhancer binding proteins (C/EBPs), fatty acid synthetase (FAS) and fatty acid-binding protein 4 (FABP4), during the adipogenesis differentiation of AMSCs. In addition, both two type of apoA1 could significantly up-regulate the intracellular gene and protein expression level of sortilin.
CONCLUSIONS ApoA1 could inhibit adipogenesis differentiation of human AMSCs and prevents obesity, and the function did not depend on the property of apoA1 mediating cholesterol efflux. The present study also provides insight into the mechanisms underlying the anti-obesity activity of apoA1 and suggests that apoA1 has the potential to prevent obesity by acting on pre-adipocytes.
GW31-e0779
Xueju Yu1,2, Fengyao Liu1,2, Yuting Liu1,2, Bingqing Bai2, Han Yin1,2, Haochen Wang1,2, Yingqing Feng1,2, Huan Ma2, Qingshan Geng1
1Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, School of Medicine, South China University of Technology, Guangzhou, China
2Department of Cardiology, Guangdong Cardiovascular Institute, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China
OBJECTIVES MicroRNAs (miRs) regulate diverse biological functions in both normal and pathological cellular conditions by post-transcriptional regulation of various genes expression. Nevertheless, the role of miRs in regulating the protective functions of omega-3 fatty acid in relation to hypoxia in cardiomyocytes, remains unknown. The aim of this study was to investigate the effects of omega-3 fatty acid supplementation on cardiomyocyte apoptosis and further, delineate the mechanisms underlying microRNA-210 (miRNA-210)-induced cardiomyocyte apoptosis in vitro.
METHODS H9C2 cultured cells were first subjected to hypoxia followed by a subsequent treatment with main component of the Omega-3 fatty acid, Docosahexaenoic Acid (DHA). Cell apoptosis were detected by flow cytometry and the expression of miR-210-3p were detected by RT-qPCR and caspase-8-associated protein 2 (CASP8AP2) at protein levels by immunoblotting. Dual luciferase assay was used to verify the mutual effect between miR-210-3p and the 3'-untranslated region (UTR) of CASP8AP2 gene.
RESULTS DHA was shown to reduce apoptosis in H9C2 cells subjected to hypoxia. Whilst DHA caused a significant increase in the expression of miR-210-3p, there was a marked reduction in the protein expression of CASP8AP2. MiR-210-3p and CASP8AP2 were significantly increased in H9C2 cardiomyocyte subjected to hypoxia. Overexpression of miR-210-3p could ameliorate hypoxia-induced apoptosis in H9C2 cells. MiR-210-3p negatively regulated CASP8AP2 expression at the transcriptional level. Both miR-210-3p mimic and CASP8AP2 siRNA could efficiently inhibit apoptosis in H9C2 cardiomyocyte subjected to hypoxia.
CONCLUSIONS We provide strong evidence showing that Omega-3 fatty acids can attenuate apoptosis in cardiomyocyte under hypoxic conditions via the up-regulation of miR-210-3p and targeting CASP8AP2 signalling pathway.
GW31-e0785
Jiaqi Yang, Yujie Zhou
Beijing Anzhen Hospital
OBJECTIVES Atherosclerosis is the main cause of cardiovascular and cerebrovascular diseases. In advanced atherosclerotic plaque, macrophage apoptosis coupled with inflammatory cytokine secretion promotes the formation of necrotic cores. Further study demonstrated that MALAT1 inhibited the expression of MAPK and NF-κB (p65) by upregulating SIRT1.
METHODS ox-LDL has been used to incubate human myeloid leukemia mononuclear cells (THP-1)-derived macrophages to establish an in vitro foam cell model. Quantitative reverse-transcription polymerase chain reaction and Western blot analyses confirmed the increased expression level of MALAT1 and the autophagy-related protein microtubule-associated protein light chain 3 (LC-3), beclin-1. The small interfering RNA study showed a significant decrease in autophagy activity and an increase in apoptotic rate when knocking down MALAT1.
RESULTS It has been demonstrated that the long-noncoding Ribo-nucleic Acid (lnc RNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), with its potent function on gene transcription modulation, maintains oxidized low-density lipoprotein (ox-LDL)-induced macrophage autophagy (i.e., helps with cholesterol efflux). It also showed that MALAT1 activated Sirtuin 1 (SIRT1), which subsequently inhibited the mitogen-activated protein kinase (MAPK) and nuclear factor kappa-B (NF-κB) signaling pathways.
CONCLUSIONS We conducted this in-vitro study to investigate the effect of MALAT1 on autophagy, apoptosis, and the formation of foam cells derived by macrophages. The primary finding indicated that knocking out MALAT1 suppressed autophagy, and thus aggravated apoptosis of macrophages through the SIRT1/MAPK/NF-κb pathway, which accelerated the progression of atherosclerosis.
GW31-e0797
Jinhua Wu, Hesong Zeng
Division of Cardiology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
OBJECTIVES Pulmonary arterial hypertension (PAH) is a progressive disorder in which endothelial dysfunction and vascular remodeling result in small pulmonary arteries occlusion, leading to right heart failure and death. Despite advances in our understanding of the pathophysiology and the management of PAH, effective treatment for this life-threatening disease is still lacking. In the current study, we aimed to explore how complement system gets involved in the pathophysiological process of PAH and test the effect of CP40-KK, a newly identified analog of selective complement C3 inhibitor CP40, on a monocrotaline (MCT) induced rat PAH model.
METHODS After MCT with or without CP40-KK treatment, right ventricle systolic pressure (RVSP) of the experimental rats was assessed by a pressure transducer system, then the animals were euthanized and lungs and hearts were dissected for further evaluation. The dry weights of the right ventricle (RV) and left ventricle (LV)+septum (S) were measured to calculate the RV/(LV+S) ratio and the RV/body weight ratio (RV/BW). The morphological indices were evaluated by hematoxylin and eosin (H&E) and immunohistochemical staining. The levels of NLRP3 inflammasome complexes and inflammatory cytokines were determined using western blotting, immunohistochemical staining and enzyme-linked immunosorbent assay (ELISA). Survival was evaluated by Kaplan–Meier analysis.
RESULTS Complement component C3 deposition increased significantly in the pulmonary small artery of the MCT group in compared with the control group, and it is associated with more perivascular monocyte/macrophage infiltration, elevated NLPR3 inflammasome activation and proinflammatory cytokines (IL-1β, IL-6 and IL-18) release, augmented vascular smooth cell proliferation and worsened hemodynamic and morphological indices, such as RVSP, RV/(LV+S) ratio, RV/BW ratio and pulmonary parietal wall thickness index, finally reduced survival rate, whereas CP40-KK treatment could significantly reverse these indices in an established rat PAH model.
CONCLUSIONS Our results indicated that complement component C3 could activate the NLRP3 inflammasome and promote subsequent release of the downstream proinflammatory cytokines, contributing to the pathophysiological process of PAH. Moreover, we found that CP40-KK treatment was protective in an established rat PAH model, which might serve as a therapeutic option for PAH.
GW31-e0798
Xiaodan Zhong, Hongjie Wang, Hesong Zeng
Department of Cardiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology
OBJECTIVES This study intends to explore whether aPC plays a protective role in the occurrence and development of diabetic cardiomyopathy and whether YB-1 is involved in its molecular mechanism.
METHODS Mice diabetic cardiomyopathy was induced by continuous intraperitoneal administration of low-dose streptozotocin (STZ), following an exogenous PC intervention treatment. The cardiac function was measured by echocardiography and invasive hemodynamics. The expression level of YB-1 was determined by Western blot and immunohistochemistry (IHC) staining. Furthermore, mice with low expression level of YB-1 were obtained by administrating YB-1 shRNA adenoviruses via tail vein and the same PC intervention was performed. Next, CoIP was used to determine the ubiquitination modification of YB-1. Finally, the signal transduction pathway of aPC was identified by specific receptor agonists and blocking antibodies.
RESULTS In diabetic cardiomyopathy mice, endogenous aPC levels were reduced. After exogenous PC intervention, PC was activated and the plasma aPC levels were increased. Meanwhile, the cardiac function of diabetic mice was significantly improved with PC intervention. The expression level of YB-1 in diabetic cardiomyopathy mice was significantly decreased. Furthermore, it was observed that the therapeutic effect of aPC disappeared in diabetic mice with low YB-1 expression. The level of ubiquitination of YB-1 with high glucose treatment was increased and the interaction between YB-1 and deubiquitinating enzyme otubain-1 (OTUB1) decreased, which were both reversed by aPC. The ubiquitination of YB-1 induced by high glucose was reduced by the overexpression of OTUB1, while knocking down the expression level of OTUB1 via shRNA increasing it. Finally, PAR1 and EPCR receptors that aPC depended to transmit extracellular signals to the intracellular regulated the expression levels of OTUB1 and YB-1.
CONCLUSIONS In conclusion, aPC protects against diabetic cardiomyopathy via regulating the ubiquitination of YB-1. The underlying mechanism is that aPC acts through PAR1 and EPCR to restrict the ubiquitination and subsequent proteasomal degradation of YB-1 by maintaining the expression of OTUB1 in cardiomyocytes under diabetic state. This study identified the protective effect of aPC on diabetic cardiomyopathy, and discussed the role of YB-1 in the occurrence and development of it, which might provide a new therapeutic strategy for diabetic cardiomyopathy.
GW31-e0799
Yuanchao Jin, Hesong Zeng, Ding Hu
Division of Cardiology, Tongji Hospital of HUST
OBJECTIVES Owing to explaining by a single race and prompting whether aneurysm is in common with the pathological changes of genetic characteristics, we conducted a meta-analysis to systematically summarize and clarify the association between single nucleotide polymorphisms (SNPs) on chromosome 9p21 and vascular aneurysm, meanwhile assessing the population attributable risk (PAR) of these variants.
METHODS A systematic search of studies on the association of the single nucleotide polymorphisms on chromosome 9p21 and vascular aneurysm was conducted in PubMed, EMBASE, China National Knowledge Infrastructure (CNKI), Chinese Biomedical (CBM) and Wan Fang (Chinese) database.
RESULTS The search yielded 14 studies including 62,805 participants. In 6 studies analyzing the association between SNPs on chromosome 9p21 and aortic aneurysm, the random effects summary estimate showed a positive relationships identified in allele model (OR: 1.36; 95% CI: 1.28–1.44; P<0.001). In 9 studies analyzing the relationships between Chr9p21 polymorphisms and intracranial aneurysm, the positive relationships were identified in allele model, with combined OR=1.28 (95% CI: 1.24–1.31; P<0.001). Similar results were found in subgroup analysis.
CONCLUSIONS SNPs with the 9p21 locus were strongly associated with the risk of vascular aneurysm.
GW31-e0823
Zhuang Ma, Hao Zhang
State Key Laboratory of Organ Failure Research, Department of Cardiology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China
OBJECTIVES Mitochondrial quality control is critical for the development of myocardial hypertrophy, in which insufficient energy supply of mitochondria plays an important role. Studies have shown that PGC-1α is closely related to energy metabolism, but previous studies on PGC-1α have different conclusions on myocardial hypertrophy. Recent studies have proved that PINK1 degrades rapidly in normal mitochondria, but accumulates in damaged mitochondria and initiates mitochondrial autophagy to clear damaged mitochondria. However, it is not clear whether PINK1 can mediate isoproterenol (Iso)-induced mitochondrial autophagy and whether the comprehensive strategy of coordinating mitochondrial autophagy and biogenesis is useful for Iso-induced cardiomyocyte injury. The changes of PINK1 and autophagy activity were measured in the model of cardiomyocyte injury induced by Iso. The effects of regulation mitochondrial autophagy and biogenesis on myocardial mitochondrial structure and function and its mechanism were explored in a cardiac hypertrophy model.
METHODS Primary rat cardiomyocytes were extracted and stimulated with Iso (10 μM) for 48 hours to construct a model of cardiomyocytes injury. The role of PINK1 in cardiomyocyte injury model was studied by adenovirus-mediated PINK1 overexpression. Then PGC-1α was activated by Metformin, and the damage indexes of reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and cardiomyocytes apoptosis were detected in each group. Protein levels of PINK1, PGC-1α, TFAM and NRF1 were detected by immunoblotting. Mitochondrial respiratory function was measured by oxygen consumption rate.
RESULTS (1) The effects of overexpression of PINK1 on cardiomyocyte function and expression of downstream gene: Compared with AD-Control+Iso group, the level of MMP increased, autophagy markers were up-regulated, ROS level and apoptosis rate decreased. Meanwhile, cell viability, ATP synthesis and mitochondrial respiratory function were improved. (2) Metformin can increase the expression of PGC-1α, which increased gradually with the stimulation concentration of Metformin to a certain extent. Compared with Iso+PINK1 group, although overexpression of PGC-1α, NRF1, TFAM and mitochondrial biogenesis, the synthesis of ATP and respiratory function of mitochondria were not significantly enhanced. (3) The effects of overexpression of MFN2 on cardiomyocyte function while overexpression the level of PINK1 and PGC-1α protein: Overexpression of MFN2 could increased mitochondrial fusion while increasing mitochondrial autophagy and regeneration in the model of cardiomyocyte injury induced by Iso. Compared with Iso+PINK1 group, ROS level and apoptosis rate decreased, cell viability, ATP synthesis and mitochondrial respiratory function were further improved.
CONCLUSIONS Increasing mitochondrial autophagy and regeneration-promoting mitochondrial fusion and improving mitochondrial quality control can reduce cardiomyocyte injury and improve capacity supply. This findings may provide a novel therapeutic strategy into the prevention of cardiomyocytes injury.
GW31-e0824
Jiayu Diao1, Hongmou Zhao2, Gong Cheng1, Penghua You1, Xiling Shou1
1Shaanxi Provincial People’s Hospital
2Xi’an Honghui Hospital
OBJECTIVES To evaluate the effect of miR-148a on CRT expression and mitochondrial function in cardiomyocytes incubated with high-glucose.
METHODS miR-148a minic and inhibitor were used to intervene the H9c2 cardiomyocytes of rats. Western-blot was used to detect the expression of CRT protein. Then the cells were divided into control group, high-glucose group (HG), HG+miR-148a minic group, HG+miR-148a minic+TG (thapsigargin, CRT agonist) group, HG+miR-148a inhibitor group, and HG+miR-148a inhibitor+CRT- (CRT-siRNA) group. The content of adenosine triphosphate (ATP) and the level of reactive oxygen species (ROS) were detected by fluorescent enzyme labeling, the activity of mitochondrial respiratory chain complex enzyme was detected by spectrophotometry, and the apoptotic rate was detected by flow cytometry.
RESULTS miR-148a minic significantly inhibited the expression of CRT protein in cardiomyocytes, and miR-148a inhibitor increased the expression of CRT. miR-148a minic inhibited the decrease of ATP production, the increase of ROS production and cell apoptosis, and the inactivity of mitochondrial respiratory chain complex enzyme in cardiomyocytes induced by high-glucose, while TG weakened the above effects of miR-148a minic. miR-148a inhibitor aggravates the mitochondrial injury and apoptosis of cardiomyocytes induced by high-glucose, while the effects of miR-148a inhibitor were partially blocked by CRT-siRNA.
CONCLUSIONS miR-148a negatively regulated the expression of CRT in cardiomyocytes, and protected the mitochondrial injury and apoptosis induced by high-glucose through inhibiting CRT.
GW31-e0826
Jiayu Diao1, Hongmou Zhao2, Gang Fan3, Xiling Shou1
1Shaanxi Provincial People’s Hospital
2Xi’an Honghui hospital
3The Second Hospital Affiliated by Xi’an Jiaotong University
OBJECTIVES This study was to evaluate the role of mitophagy in the protective effect on high glucose-induced hypertrophy of RES in H9c2 cardiomyocyte.
METHODS The level of mitophagy in rats and H9c2 cells was detected. The reactive oxygen species (ROS) production, mitochondrial membrane potential (MMP), the activation of mitochondrial permeability transition pore (MPTP) and apoptosis rate, and indicators of cardiomyoctye hypertrophy were measured in each group.
RESULTS The expression of Parkin and LC3II/LC3I were significantly decreased in DCM rats and H9c2 cells incubated with high-glucose, and were both raised up by RES pretreatment. RES inhibited the production of ROS, dissolved MMP, activation of MPTP and excessive apoptosis. And the protective effects of RES were all reversed by Parkin-siRNA. RES alleviated HG-induced cardiomyocyte hypertrophy, the effect of RES was also blocked by Parkin-siRNA.
CONCLUSIONS Parkin-mediated mitophagy was involved in the protective effects of RES on the oxidative stress and cardiomyocyte hypertrophy.
GW31-e0832
Ye Chang, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES This study aimed to explore whether angiotensin II (Ang II) inhibits the proliferation of human umbilical vein endothelial cells (HUVECs) by changing mitochondrial energy metabolism, and whether atorvastatin has a protective role via restoration of endothelial function.
METHODS HUVECs were treated with 1 μM Ang II alone or with 10 μM atorvastatin for 24 h. Proliferation was detected by MTT assay, cell counting, 5-ethynyl-2ʹ-deoxyuridine assay and real-time cell analyzer. Mitochondrial energy metabolism including oxygen consumption rate and extracellular acidification rate were measured using a Seahorse metabolic flux analyzer. Mitochondrial membrane potential was detected under fluorescence microscope following staining with tetramethylrhodamine. Respiratory chain complexes I–V were detected using western blotting.
RESULTS The current study showed that Ang II inhibits the proliferation of HUVECs. Results from the Seahorse metabolic flux analyzer indicated that Ang II decreased basal oxygen consumption, maximal respiration capacity, spare respiration capacity, adenosine triphosphate-linked respiration and non-mitochondrial respiration. By contrast, Ang II increased the proton leak. Additionally, Ang II increased glycolysis, glycolytic capacity and non-glycolytic acidification. Furthermore, these effects were all suppressed by atorvastatin.
CONCLUSIONS The results indicated that atorvastatin prevents cellular energy metabolism switching from oxidative phosphorylation to glycolysis induced by Ang II and protected the proliferative ability of HUVECs.
GW31-e0834
Ye Chang, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES This study was aimed to evaluate lysosomes-mitochondria cross-signaling in angiotensin II (Ang II)-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and whether atorvastatin played a protective role via lysosomal-mitochondrial axis.
METHODS Apoptosis was detected by flow cytometry, Hoechst 33342 and AO/EB assay. The temporal relationship of lysosomal and mitochondrial permeabilization was established. Activity of Cathepsin D (CTSD) was suppressed by pharmacological and genetic approaches. Proteins production were measured by western blotting.
RESULTS Our study showed that Ang II could induce the apoptosis of HUVECs in a dose-depended and time-depended manner. Exposure to 1 μM Ang II for 24 h resulted in mitochondrial depolarization, cytochrome c release, and increased ROS production. Lysosomal permeabilization and CTSD redistribution into the cytoplasm occurred several hours prior to mitochondrial dysfunction. These effects were all suppressed by atorvastatin. Either pharmacological or genetic inhibition of CTSD preserved mitochondrial function and decreased apoptosis in HUVECs. Most importantly, we found that the protective effect of atorvastatin was significantly greater than pharmacological or genetic inhibition of CTSD. Finally, overexpression of CTSD without exposure to Ang II had no effect on mitochondrial function and apoptosis.
CONCLUSIONS Our data strongly suggested that Ang II induced apoptosis through the lysosomal-mitochondrial axis in HUVECs. Furthermore, atorvastatin played an important role in the regulation of lysosomes and mitochondria stability, resulting in an antagonistic role against Ang II on HUVECs.
GW31-e0859
Meng Li1, Junping Zhang1, Zhongwen Qi2, Ke Zhu2
1First Teaching Hospital of Tianjin University of Traditional Chinese Medicine
2Tianjin University of Traditional Chinese Medicine
OBJECTIVES Observed the effect of Si-Miao-Yong-An decoction on AS vulnerable plaques, and the vasa vasorum (VV) angiogenesis and maturation were as the entry point to further discuss the mechanism of effect.
METHODS The male ApoE-/- mice were randomized into 3 groups: model group, simvastatin group and Si-Miao-Yong-An group, and C57BL/6 mice were used as the control group. The ApoE-/- mice were fed with high-fat diet added 1.1% L-methionine for 8 weeks to establish the AS vulnerable plaque model, and the C57BL/6 mice were as control group. After 8 weeks, the pathological morphology of plaque was observed by HE staining; the VV density in plaque and aortic adventitia were observed by immunohistochemistry; VV maturation was measured by double-labelling immunofluorescence; the critical proteins of HIF-1α-Apelin/APJ and Ang-1/Tie signal pathways were detected by western blotting.
RESULTS Si-Miao-Yong-An decreased the plaque area and the ratio of plaque area and lumen area, increased the minimum thickness of fibrous cap, which significantly improved the pathological feature of aortic plaque in mice; it effectively suppressed the VV neovascularization; promoted smooth muscle cells recruitment; it regulated the HIF-1α-Apelin/APJ and Ang-1/Tie signal pathways.
CONCLUSIONS Si-Miao-Yong-An regulated the HIF-1α-Apelin/APJ signal pathway, suppressing VV neovascularization; it also regulated the Ang-1/Tie signal pathway, promoting VV maturation, which promoted the VV network reconstruction, improved VV function and finally stabilized AS vulnerable plaque.
GW31-e0860
Qiuyu Li1, Pengli Zhu1,2
1Department of Geriatric Medicine, Fujian Provincial Hospital 350001
2Fujian Provincial Center of Geriatrics; Fujian Provincial Key Laboratory of Geriatric Disease 350001
OBJECTIVES To explore the effects of Apelin on D-galactose (D-Gal)-induced cardiomyocyte senescence and mitochondrial energy metabolism.
METHODS Through electroporation transfection technique, the Apelin receptor over and low expression (h-APJ and Si-APJ) H9c2 cells were established. To create the aging model, H9c2 cells were intervened with D-Gal in an optimum concentration of 20 g/L and tested by using cell counting kit-8, beta-galactosidase staining, RT-PCR detect P16 gene expression to confirm the aging myocardial model was established successfully. These cells were divided into six experimental groups: (1) Control Group: H9c2 cells were cultured without special treatment; (2) Aging Group: H9c2 cells were cultured with D-Gal; (3) Group Si-APJ D: Si-APJ H9c2 cells were cultured with D-Gal; (4) Group Si-APJ DA: Si-APJ H9c2 cells were cultured with D-Gal and Apelin; (5) Group h-APJ D: h-APJ H9c2 cells were cultured with D-Gal; (6) Group h-APJ DA: h-APJ H9c2 cells were cultured with D-Gal and Apelin. Using the high-content imaging analysis system to detect the fluorescence intensity, the mitochondrial morphology was analyzed including its density, size, and texture parameter. The citrate synthase detection kit detects cell citrate synthase activity. Western-Blotting detects protein expression of aging marker P16 and PGC-1α, fatty acid metabolism related marker CPT1A, glucose metabolism marker GLUT4. RT-PCR detects associated gene expression of P16, CPT1A, GLUT4, TFAM, NRF1.
RESULTS (1) H9c2 cells in the Aging Group compared with the Control Group, the relative expression levels of P16 increased; whereas in the h-APJ cells, with the intervention of Apelin, its expression decreased; besides it is no significant difference in Apelin receptor low expressing group, including Si-APJ D and Si-APJ DA. (2) The cell in the Aging Group showed significantly lower citrate synthase activity than the Control Group; the citrate synthase activity in the h-APJ DA Group was higher than that in the h-APJ D Group; while it was only slightly higher in Si-APJ DA Group than that of Si-APJ D Group. (3) Compared with the Control Group, the cell relative expression of PGC-1α, GLUT4 and CPT1A were reduced in the Aging Group; in h-APJ cells, the relative expression of these three markers consistently increased when they were in existence of Apelin; however there was no significant difference of them in between Si-APJ DA Group and Si-APJ D Group. 4. H9c2 cells in the Aging Group, compared with the Control Group, their mitochondrial biosynthesis decreased, but their size and texture parameters did not change significantly; in the h-APJ cells, with or without the intervention of Apelin, there was no obvious change in cell mitochondrial structure, membrane potential, and biosynthesis; furthermore it showed similar manifestation in Si-APJ cells.
CONCLUSIONS Apelin has a protective effect on D-Gal induced cardiomyocyte senescence. Apelin has a protective effect on mitochondrial energy metabolism in senescent cardiomyocytes induced by D-Gal, but it has no significant effect on mitochondrial structural damage and mitochondrial biosynthesis.
GW31-e0865
Peining Liu, Wen Xi, Junhui Liu, Yuanyuan Wei, Zuyi Yuan, Yue Wu
The First Affiliated Hospital of Xi’an Jiaotong University
OBJECTIVES Recently the Canakinumab Anti-inflammatory Thrombosis Outcome Study (CANTOS) has confirmed the beneficial effect of inhibition of interleukin (IL)-1β on major cardiovascular events. However, there exists efficacy discrepancy depending on the reduction of serum high-sensitivity C-reactive protein (CRP). This study aimed to explore mechanisms underlying this discrepancy in a proper animal model for identification of individuals most likely to benefit from Canakinumab treatment.
METHODS Cynomolgus monkeys consumed a chow (n=20) or atherogenic diet (n=60) for over 3 years. Blood was collected after an overnight fasting. Serum levels of triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL-c) and glucose were measured in the central laboratory using an automatic biochemical analyzer. Serum levels of CRP, IL-6 and IL-1β were assayed by enzyme-linked immunosorbent assay (ELISA). Intima-media thickness (IMT) of common carotid arteries was measured by ultrasound. Monkeys were sedated by an intramuscular injection of 10% ketamine hydrochloride after fasting overnight and placed in a supine position with the neck fully exposed. Ultrasound examinations were performed with the use of a 15-MHz ultrasonic probe by a single trained sonographer who was kept blinded from the trial information. Whenever an atherosclerotic plaque was present, it was further confirmed by color Doppler ultrasonongraphy examination and included in the measurements of IMT.
RESULTS The atherogenic diet significantly raised the serum TC, LDL-c and carotid IMT both in male and female monkeys. Meanwhile, the serum levels of CRP and IL-1β were increased only in female atherogenic diet group compared with that of the female chow diet group. Correlation analysis showed that the levels of CRP and IL-1β were positively correlated with IMT in females. While in males, only IL-6 level was positively correlated with IMT. Association analysis between these circulating inflammatory cytokines and traditional risk factors of atherosclerosis depicted that IL-6 levels were positively correlated with fasting blood glucose, TC, LDL-c and TC/HDL-c ratio in males; IL-1β levels were positively correlated with fasting blood glucose in males. Whereas in females, only the significant associations between IL-6 and glucose were observed, indicating that glucose and lipid metabolism may have influences on systematic inflammatory status. When divided into two groups based on IMT median, IL-1β was showed to be an independent risk factor for higher IMT after adjusted for traditional risk factors including age, BMI, fasting blood glucose, TG and TC/HDL ratio in female monkeys (OR: 1.625, 95% CI: 1.049–2.516 for per 0.1 ng/mL rise). Additionally, in both sexes, IL-6 was positively and significantly correlated with IL-1β. Besides, IL-6 was also positively and significantly correlated with CRP in female monkeys. These results lent a support to the existence of CRP/IL-6/IL-1β axis and its possible roles in atherosclerosis, especially in females.
CONCLUSIONS Serum IL-1β elevation is independently associated with increased carotid IMT in female Cynomolgus monkeys, suggesting a promising animal model for further studies on the pathological roles of IL-1β/IL-6/CRP axis in atherosclerosis.
GW31-e0868
Lele Cheng, Zuyi Yuan
The First Affiliated Hospital of Xi’an Jiaotong University
OBJECTIVES Recent studies reveal that bile acid metabolite composition and their metabolism are changed in metabolic disorders, such as obesity, type 2 diabetes, and metabolic associated fatty liver disease (MAFLD). Glycoursodeoxycholic acid (GUDCA), glycine-conjugated bile acid produced from intestinal bacteria, may play a role in metabolic disorders, yet the mechanism remains largely unknown.
METHODS Metabolomic analysis of 163 serum and stool samples of our metabolic disease cohort was performed and comparative analysis was conducted between high and low glycated hemoglobin A1c (HbA1c) level groups. High-throughput RNA-sequencing analysis of liver transcriptome of high-fat diet (HFD)-fed mice was conducted. The effects of GUDCA on body weight, glucose homeostasis, insulin sensitivity, hepatic steatosis, lipid profiles, endoplasmic reticulum (ER) homeostasis and apoptosis were assessed.
RESULTS GUDCA levels were decreased in both serum and stool samples from patients with glucose metabolic disorders. RNA-sequencing results indicated that GUDCA alleviated ER stress in livers of HFD-fed mice without alteration of liver metabolism. In vitro, GUDCA reduced palmitic acid induced-ER stress and -apoptosis, as well as stabilized calcium homeostasis. In vivo, GUDCA exerted similar effects as tauroursodeoxycholic acid (TUDCA) on amelioration of HFD-induced insulin resistance and hepatic steatosis. In parallel, ER stress and apoptosis were decreased in GUDCA-treated mice as compared to vehicle-treated mice.
CONCLUSIONS Reduced GUDCA is an indicator of metabolic disorders. Supplementation of GUDCA could be an option for the treatment of metabolic disorders, including insulin resistance and hepatic steatosis.
GW31-e0881
Xianchun Yan1, Jiaxing Sun1,2, Ziyan Yang1, Peiran Zhang1, Liang Liang1, Hua Han1
1State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, Fourth Military Medical University
2Department of Ophthalmology, Eye Institute of Chinese PLA, Xijing Hospital, Fourth Military Medical University
OBJECTIVES After active embryonic and early postnatal vascularization mediated primarily by angiogenesis, endothelial cells (ECs) in most tissues adopt a quiescent state, which is critical for tissue perfusion and EC functions. Notch signaling is essential in maintaining EC quiescence, but the downstream mechanisms have been elusive.
METHODS EdU or propidium iodide (PI) incorporation assay was used to determine EC proliferation ability. Lumen formation and fibrin beads sprouting assays were employed to evaluate EC angiogenic ability in vitro. Immunostaining of retinas was used to observe angiogenesis in vivo. RNA-seq was used to compare gene expression profiles. qRT-PCR and western blotting were used to determine gene expression level.
RESULTS miR-218 is a novel downstream Notch effector in quiescent ECs. Notch activation upregulated, while Notch blockade downregulated, miR-218 and its host gene Slit2, likely via transactivating the Slit2 promoter. miR-218 overexpression in human umbilical vein endothelial cells (HUVECs) significantly repressed cell proliferation and eroded sprouting in vitro. Transcriptional profiling showed that miR-218 overexpression attenuates MYC transcription program. MYC overexpression rescued miR-218-mediated repression of proliferation and sprouting of HUVECs. Furthermore, miR-218 downregulates MYC via multiple mechanisms including reducing MYC mRNA by unidentified target(s), repressing MYC translation by targeting hnRNPA1, and promoting MYC degradation by targeting EYA3. Inhibition of miR-218 partially reversed Notch activation-induced repression of HUVEC proliferation and sprouting. In vivo, intravitreal injection of miR-218 reduced retinal EC proliferation, and attenuated pathological angiogenesis in a choroidal neovascularization (CNV) model.
CONCLUSIONS miR-218 mediates the effect of Notch activation on promoting EC quiescence via MYC, and holds potentials in the treatment of angiogenesis-related diseases.
GW31-e0886
Shasha Yu, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES Previous studies have demonstrated that angiotensin II (Ang II) is involved in the process of atherosclerosis and vascular restenosis through its proinflammatory effect. Bcl-2-associated athanogene 3 (BAG3) had been suggested to be associated with proliferation, migration and invasion in many types of tumor. However, the role of BAG3 among the proliferative process of vascular smooth muscle cells (VSMCs) induced by Ang II, to the best of our knowledge, remains to be investigated. The present study demonstrated that in growth-arrested VSMCs, Ang II-induced VSMC proliferation, accompanied by increased BAG3 mRNA and protein expression levels in a dose- and time-dependent manner. BAG3 expression levels were measured in VSMCs treated in the presence or absence of Ang II.
METHODS The proliferation of VSMCs was assessed using manual cell counting and Cell Counting kit-8 assays. mRNA and protein expression levels of BAG3, Toll-like receptor 4 (TLR4), proliferating cell nuclear antigen, nuclear factor (NF)-κB p65, smooth muscle protein 22α and phosphorylated NF-κB p65 were assessed by reverse transcription-quantitative polymerase chain reaction and western blotting, respectively.
RESULTS In non-transfected or scramble short hairpin RNA (shRNA)-transfected VSMCs cells, Ang II significantly induced VSMC proliferation. However, this Ang II-induce proliferation was attenuated when BAG3 was silenced, suggesting that inhibition of BAG3 may somehow reduce proliferation in Ang II-induced VSMCs. Furthermore, the TLR4/NF-κB p65 signaling pathway was involved in BAG3 gene upregulation.
CONCLUSIONS In conclusion, to the best of our knowledge, the present study demonstrated for the first time that inhibition of BAG3 attenuates cell proliferation. Furthermore, Ang II induced VSMCs proliferation through regulation of BAG3 expression via the TLR4/NF-κB p65 signaling pathway.
GW31-e0890
Shasha Yu, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES It has been proven that phenotype shifting, from the contractile phenotype to the synthetic phenotype, of vascular smooth muscle cells (VSMCs), plays an important role in vascular diseases such as atherosclerosis, restenosis, and hypertension. Recently, accumulating evidence suggests that Klotho is associated with many cardiovascular diseases or damage. Through the estimation of the proliferation and migration of Ang II-induced VSMCs and the related intracellular signal transduction pathways, we researched the effects of Klotho on phenotype modulation in this study.
METHODS A rat vascular smooth muscle cell line was grown in vitro with or without Ang II or Klotho, and cell proliferation and migration were evaluated.
RESULTS The dose-dependent inhibition of Ang II-induced proliferation and migration by Klotho was shown in VSMCs. The phenotype modulation was inhibited by Klotho co-treatment; this co-treatment promoted the expression of contractile phenotype marker proteins, including SM22α, and also the proliferation phenotype marker protein PCNA compared with Ang II alone, which was suppressed, and activated VSMCs. Furthermore, by reducing the expression of G0/G1-specific regulatory proteins such as cyclin D1, cyclin-dependent kinase (CDK) 4, cyclin E, and CDK2, cell cycle arrest was induced by Klotho at G0/G1 phase. Although Ang II strongly stimulated NF-κB, p65, Akt, and ERK phosphorylation, these activation events were diminished by co-treatment with Ang II and Klotho.
CONCLUSIONS Phenotype modulation of Ang II-induced VSMCs and stimulation of the NF-κB, p65, Akt, and ERK signaling pathways were inhibited by Klotho, which suggests that Klotho may play an important role in the phenotype modulation of VSMCs.
GW31-e0892
Naijin Zhang, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES SIRT1 and STAT3 are key to human aortic vascular smooth muscle cells (HAVSMCs) proliferation, migration and phenotypic transformation, but the regulatory mechanism of SIRT1-STAT3 in this process is still unclear. Septin4 is a cytoskeleton-related protein that regulates oxidative stress-vascular endothelial injury. However, the role and underlying mechanism of Septin4 in atherosclerosis remains unknown. Here, we revealed the role and mechanism of Septin4 in regulating SIRT1-STAT3 in atherosclerosis. We determined that the expression of Septin4 were markedly increased in Apoe(-/-) atherosclerosis mice and PDGF-BB-induced HAVSMCs.
METHODS Knockdown of Septin4 significantly increased PDGF-BB-induced HAVSMCs proliferation, migration and phenotypic transformation, while overexpression of Septin4 had the opposite effects.
RESULTS Mechanically, co-immunoprecipitation results demonstrated that Septin4 was a novel interacting protein of STAT3 and SIRT1. Septin4 formed a complex with SIRT1-STAT3, enhancing the interaction between SIRT1 and STAT3, ensuing promoting SIRT1-regulated STAT3-K685 deacetylation and STAT3-Y705 dephosphorylation, which inhibited PDGF-BB-induced HAVSMCs proliferation, migration and phenotype transformation.
CONCLUSIONS Therefore, our findings provide novel insights into the prevention and treatment of atherosclerosis.
GW31-e0895
Naijin Zhang, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES Oxidative stress induced vascular endothelial cell injure is one of the key and initial event in the development of atherosclerosis. Septin4, as a member of GTP binding protein family, is widely expressed in the eukaryotic cells and considered to be an essential component of the cytoskeleton which is involved in many important physiological processes. However, whether Septin4 is involved in cardiovascular diseases, such as oxidative stress inducted endothelial cell injury still unclear.
METHODS PARP1 as a DNA repair enzyme can be activated by identifying DNA damaged fragments, which consumes high levels of energy and leads to vascular endothelial cell apoptosis.
RESULTS Here, our results first found that Septin4 is involved in oxidative stress induced endothelial cell ROS production and apoptosis through knock-down and over-expression Septin4 approaches. Furthermore, to explore how Septin4 is involved in oxidative stress induced endothelial cells injure, we first identified that Septin4 is a novel PARP1 interacting protein and the interaction is enhanced under oxidative stress.
CONCLUSIONS In conclusions, our founding indicates that Septin4 is a novel essential factor involved in oxidative stress induced vascular endothelial cell injury by interacting with apoptosis-related protein PARP1.
GW31-e0896
Naijin Zhang, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES The elevated expression of poly(ADP-ribose) polymerase-1 (PARP1) and increased PARP1 activity, namely, poly(ADP-ribosyl)ation (PARylation), have been observed in cardiac remodeling, leading to extreme energy consumption and myocardial damage. However, the mechanisms underlying the regulation of PARP1 require further study. WWP2, a HECT-type E3 ubiquitin ligase, is highly expressed in the heart, but its function there is largely unknown. Here, we clarified the role of WWP2 in the regulation of PARP1 and the impact of this regulatory process on cardiac remodeling.
METHODS We determined that the knockout of WWP2 specifically in myocardium decreased the level of PARP1 ubiquitination and increased the effects of isoproterenol (ISO)-induced PARP1 and PARylation, in turn aggravating ISO-induced myocardial hypertrophy, heart failure, and myocardial fibrosis. Similar findings were obtained in a model of ISO-induced H9c2 cells with WWP2 knockdown, while the reexpression of WWP2 significantly increased PARP1 ubiquitination and decreased PAPR1 and PARylation levels.
RESULTS Mechanistically, coimmunoprecipitation results identified that WWP2 is a novel interacting protein of PARP1 and mainly interacts with its BRCT domain, thus mediating the degradation of PARP1 through the ubiquitin-proteasome system. In addition, lysine 418 (K418) and lysine 249 (K249) were shown to be of critical importance in regulating PARP1 ubiquitination and degradation by WWP2.
CONCLUSIONS These findings reveal a novel WWP2-PARP1 signal transduction pathway involved in controlling cardiac remodeling and may provide a basis for exploring new strategies for treating heart disorders related to cardiac remodeling.
GW31-e0897
Naijin Zhang, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES Oxidative stress-associated endothelial injury is the initial event and major cause of multiple cardiovascular diseases such as atherosclerosis and hypertensive angiopathy. A protein homeostasis imbalance is a critical cause of endothelial injury, and homologous to E6AP C-terminus (HECT)-type E3 ubiquitin ligases are the core factors controlling protein homeostasis. Although HECT-type E3 ubiquitin ligases are involved in the regulation of cardiac development and diseases, their roles in endothelial injury remain largely unknown. This study aimed to identify which HECT-type E3 ubiquitin ligase is involved in endothelial injury and clarify the mechanisms at molecular, cellular, and organism levels. We revealed a novel role of the HECT-type E3 ubiquitin ligase WWP2 in regulating endothelial injury and vascular remodeling after endothelial injury.
METHODS Endothelial/myeloid-specific WWP2 knockout in mice significantly aggravated angiotensin II/oxidative stress-induced endothelial injury and vascular remodeling after endothelial injury. The same results were obtained from in vitro experiments.
RESULTS Mechanistically, the endothelial injury factor Septin4 was identified as a novel physiological substrate of WWP2. In addition, WWP2 interacted with the GTPase domain of Septin4, ubiquitinating Septin4-K174 to degrade Septin4 through the ubiquitin-proteasome system, which inhibited the Septin4-PARP1 endothelial damage complex.
CONCLUSIONS These results identified the first endothelial injury-associated physiological pathway regulated by HECT-type E3 ubiquitin ligases in vivo as well as a unique proteolytic mechanism through which WWP2 controls endothelial injury and vascular remodeling after endothelial injury. These findings might provide a novel treatment strategy for oxidative stress-associated atherosclerosis and hypertensive vascular diseases.
GW31-e0903
Moujie Liu, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES Post-traumatic stress disorder (PTSD) is related with myocardial injury and cardiac dysfunction, while the molecular mechanism has not been clear. This study investigated whether TLR4/MyD88/NF-κB-mediated inflammation involved in myocardial injury of PTSD.
METHODS Adult male Wistar rats were exposed to single-prolonged stress (SPS), which was used broadly as a animal model of PTSD. Morris Water Maze (MWM) test and forced swimming test (FST) was carried out for behavioral testing. The protein expression of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in the left ventricular of heart and TLR4/MyD88/NF-κB-mediated inflammation were examined.
RESULTS Our results showed that there were obvious increased in the protein expression of ANP and BNP in heart after exposure to SPS, SPS also significantly enhanced the serum level of IL-1β and TNF-α, and meanwhile, the TLR4/MyD88/NF-κB pathway were activated.
CONCLUSIONS These results demonstrated that the TLR4/MyD88/NF-κB pathway were involved in the myocardial injury of PTSD, which might be one of possible molecular mechanism contributed to the pathogenesis of cardiac dysfunction in PTSD.
GW31-e0907
Pengyu Jia, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES Agomelatine is a melatonin (MT1/MT2) receptor agonist and serotonin (5-HT2C) receptor antagonist. To study the effects of agomelatine on myocardial ischemia reperfusion injury (MIRI), an isolated rat heart model was utilized. To induce MIRI, rat hearts were isolated and subjected to 30 min of ischemia followed by 120 min of reperfusion.
METHODS Rats were intraperitoneally injected with agomelatine (10, 20 or 40 mg/kg) 1 h before heart isolation.
RESULTS Agomelatine (20 mg/kg and 40 mg/kg) significantly improved cardiac function, alleviated pathological changes in the ischemic myocardium, reduced myocardial infarct size and decreased release of creatine kinase-MB and lactate dehydrogenase. Heart tissue from agomelatine-treated rats retained higher NAD(+) content and was more resistant to Ca(2+), indicating inhibition of mitochondrial permeability transition pore (MPTP) opening.
CONCLUSIONS Notably, agomelatine’s protective effects were abrogated by atractyloside, a MPTP opener. We also found that agomelatine significantly enhanced GSK-3β phosphorylation and decreased expression of cytochrome C, cleaved caspase 9 and cleaved caspase 3, resulting in a decreased apoptosis rate. These findings demonstrate that agomelatine protects against MIRI by inhibiting MPTP opening.
GW31-e0908
Pengyu Jia, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES The increased level of saturated fatty acids (SFAs) is found in patients with diabetes, obesity, and other metabolic disorders. SFAs can induce lipotoxic damage to cardiomyocytes, but the mechanism is unclear. The long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) acts as a key regulator in palmitic acid (PA)-induced hepatic steatosis, but its role in PA-induced myocardial lipotoxic injury is still unknown. The aim of this study was to explore the role and underlying mechanism of MALAT1 in PA-induced myocardial lipotoxic injury.
METHODS MALAT1 expression in PA-treated human cardiomyocytes (AC16 cells) was detected by RT-qPCR. The effect of MALAT1 on PA-induced myocardial injury was measured by Cell Counting Kit-8, lactate dehydrogenase (LDH), and creatine kinase-MB (CK-MB) assays. Apoptosis was detected by flow cytometry. The activities of cytokines and nuclear factor (NF)-κB were detected by enzyme-linked immunosorbent assay. The interaction between MALAT1 and miR-26a was evaluated by a luciferase reporter assay and RT-qPCR. The regulatory effects of MALAT1 on high mobility group box 1 (HMGB1) expression were evaluated by RT-qPCR and western blotting.
RESULTS MALAT1 was significantly upregulated in cardiomyocytes after PA treatment. Knockdown of MALAT1 increased the viability of PA-treated cardiomyocytes, decreased apoptosis, and reduced the levels of LDH, CK-MB, TNF-α, and IL-1β. Moreover, we found that MALAT1 specifically binds to miR-26a and observed a reciprocal negative regulatory relationship between these factors. We further found that the downregulation of MALAT1 represses HMGB1 expression, thereby inhibiting the activation of the Toll-like receptor 4 (TLR4)/NF-κB-mediated inflammatory response. These repressive effects were rescued by an miR-26a inhibitor.
CONCLUSIONS We demonstrate that MALAT1 is induced by SFAs and its downregulation alleviates SFA-induced myocardial inflammatory injury via the miR-26a/HMGB1/TLR4/NF-κB axis. Our findings provide new insight into the mechanism underlying myocardial lipotoxic injury.
GW31-e0909
Xia Li, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES Oxidative stress participates in several heart diseases and is an important mechanism contributing to the pathological alterations of myocardial cell injury. In recent years, ubiquitylation has been demonstrated to be an important biochemical reaction associated with apoptosis.
METHODS To investigate the effects and interactions of the E3 ligase F-box and WD repeat domain containing 7 (Fbw7) and MCL1 apoptosis regulator, BCL2 family member (Mcl-1) in myocardial cells during oxidative stress, Cell Counting Kit-8, flow cytometry, western blot, reactive oxygen species and co-immunoprecipitation assays were conducted.
RESULTS The current study revealed that Fbw7 may facilitate apoptosis via the Mcl-1-Bax pathway in oxidative stress-induced myocardial H9c2 cell injury. Mcl-1 inhibits the functions of Bcl-2 family members, including the mitochondrial apoptosis factor Bax, to maintain cell viability; however, the present study suggested that Fbw7 may degrade Mcl-1 and impaired this process.
CONCLUSIONS Therefore, it may be hypothesized that Fbw-7 promotes myocardial cell injury via interacting with Mcl-1.
GW31-e0927
Shuang Chen, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES Abnormal proliferation of vascular smooth muscle cells (VSMCs) is a hallmark of vascular lesions, such as atherosclerosis and restenosis. PDGF-β β, an isoform of PDGF (platelet-derived growth factor), has been demonstrated to induce proliferation and migration of VSMCs. Atorvastatin calcium, a selective inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, has favorable protective effects on VSMCs. This study examined the effects of atorvastatin calcium on the proliferation and migration of PDGF-β β-treated VSMCs, as well as its underlying mechanisms.
METHODS MTT assays, Edu imaging, cell cycle analysis, wound healing assays, transwell migration assays, and western blot analysis were performed.
RESULTS Atorvastatin calcium significantly inhibited cell proliferation, DNA synthesis and cell migration of PDGF-β β-treated VSMCs. We demonstrated that atorvastatin calcium induced cell cycle arrest in the G0/G1 phase in response to PDGF-β β stimulation and decreased the expression of G0/G1-specific regulatory proteins, including proliferating cell nuclear antigen (PCNA), CDK2, cyclin D1, cyclin E and CDK4 in PDGF-β β-treated VSMCs. Moreover, pretreatment with atorvastatin calcium inhibited the PDGF-β β-treated phosphorylation of PDGFRβ and Akt, whereas atorvastatin calcium did not affect the phosphorylation of PLC-γ1 or (ERK) 1/2.
CONCLUSIONS Our data suggested that atorvastatin calcium inhibited abnormal proliferation and migration of VSMCs through G0/G1 cell cycle arrest and suppression of the PDGFRβ-Akt signaling cascade.
GW31-e0928
Jiaqi Xu, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES Vascular smooth muscle cells (VSMCs) play a crucial role in the progression of atherosclerosis. Paired box 9 (Pax9) is a member of the Pax gene family which participates in the development of various tissues and organs. However, the effect of Pax9 on atherosclerosis and VSMCs and the underlying mechanisms remain unclear.
METHODS Western blotting was performed to assess Pax9 expression in atherosclerosis and VSMCs. Pax9 siRNA and overexpression plasmid were constructed to explore the biological function. Cell proliferation assay, phalloidin staining, and Transwell assay, accompanied by the sonic hedgehog (Shh) signaling pathway antagonist, cyclopamine (5 μM) and agonist, SAG (100 nM), were used to evaluate the VSMC phenotype, proliferation, and migration, as well as explore the associated mechanisms.
RESULTS We first discovered Pax9 to be significantly increased in atherosclerotic mice and platelet-derived growth factor-BB (PDGF-BB)-induced VSMCs. Pax9 knockdown inhibited the phenotypic transformation, proliferation, and migration of VSMCs, whereas the opposite effect was observed when Pax9 was overexpressed. Next, we established that Shh was activated in PDGF-BB-induced VSMCs. Moreover, Pax9 overexpression further activated Shh and exacerbated the phenotypic transformation, proliferation, and migration of PDGF-BB-induced VSMCs. These changes were effectively inhibited by treatment with the Shh signaling pathway antagonist. Consistently, Pax9 knockdown down-regulated Shh expression and inhibited the phenotypic transformation, proliferation, and migration of PDGF-BB-induced VSMCs. Treatment with the Shh signaling pathway agonist prevented these changes.
CONCLUSIONS Pax9 regulated VSMC phenotypic transformation, proliferation, and migration via Shh, which may represent a novel target for the treatment of atherosclerosis.
GW31-e0934
Liang Guo, Yingxian Sun
The First Hospital of China Medical University
OBJECTIVES Complement component 5 (C5) has been described to play an important role in the development and progression of atherosclerosis and cardiovascular disease. Our aim was to determine whether genetic variation of C5 was associated with ischemic stroke (IS) in northeast Chinese population.
METHODS We used a case-control study involving 386 IS patients and 386 non-IS controls from a rural population and determined the genotypes of five polymorphisms (rs12237774, rs17611, rs4837805, rs7026551, and rs1017119) of C5 gene by Snapshot single-nucleotide polymorphism genotyping assays to assess any links with IS.
RESULTS In univariate analysis, rs17611 was significantly associated with IS in the additive model, the dominant model, and recessive model (additive P 0.031, dominant P 0.034, and recessive P 0.027). After adjustment for Binary Logistic Regression, rs17611 polymorphism was still significant in three models (adjusted odds ratio (OR)=1.306, 95% confidence interval (CI)=1.069–1.595, P-value=0.009 in an additive model; OR=1.378, 95% CI=1.024–1.856, P-value=0.035 in a dominant model; and OR=1.511, 95% CI=1.048–2.18, P-value=0.027 in a recessive model).
CONCLUSIONS In this sample of patients, genetic variation of rs17611 in C5 is associated with higher prevalence of IS.
GW31-e0940
Yintao Chen1,2, Yingxian Sun1
1The First Hospital of China Medical University
2The First Affiliated Hospital of Chongqing Medical University
OBJECTIVES Many clinical and experimental studies have shown that treatment with statins could prevent myocardial hypertrophy and remodeling induced by hypertension and myocardial infarction. But the molecular mechanism was not clear. We aimed to investigate the beneficial effects of atorvastatin on hypertension-induced myocardial hypertrophy and remodeling in spontaneously hypertensive rats (SHR) with the hope of revealing other potential mechanisms or target pathways to interpret the pleiotropic effects of atorvastatin on myocardial hypertrophy.
METHODS The male and age-matched animals were randomly divided into three groups: control group (8 WKY), SHR (8 rats) and intervention group (8 SHR). The SHR in intervention group were administered by oral gavage with atorvastatin (suspension in distilled water, 10 mg/kg once a day) for 6 weeks, and the other two groups were administered by gavage with equal quantity distilled water. Blood pressure of rats was measured every weeks using a standard tail cuff sphygmomanometer. Left ventricular (LV) dimensions were measured from short-axis views of LV under M-mode tracings using Doppler echocardiograph. Cardiomyocyte apoptosis was assessed by the TUNEL assay. The protein expression of C/EBPβ, PGC-1α and UCP3 were detected by immunohistochemistry or Western blot analysis.
RESULTS At the age of 16 weeks, the mean arterial pressure of rats in three groups were 103.6±6.1, 151.8±12.5 and 159.1±6.2 mmHg respectively, and there wasn’t statistically significant difference between the SHR and intervention groups. Staining with Masson’s trichrome demonstrated that the increased interstitial fibrosis of LV and ventricular remodeling in the SHR group were attenuated by atorvastatin treatment. Echocardiography examination exhibited that SHR with atorvastatin treatment showed an LV wall thickness that was obviously lower than that of water-treated SHR. In hypertrophic myocardium, accompanied by increasing C/EBPβ expression and the percentage of TUNEL-positive cells, the expression of Bcl-2/Bax ratio, PGC-1α and UCP3 were reduced, all of which could be abrogated by treatment with atorvastatin for 6 weeks.
CONCLUSIONS This study further confirmed that atorvastatin could attenuate myocardial hypertrophy and remodeling in SHR by inhibiting apoptosis and reversing changes in mitochondrial metabolism. The C/EBPβ/PGC-1α/UCP3 signaling pathway might also be important for elucidating the beneficial pleiotropic effects of atorvastatin on myocardial hypertrophy.
GW31-e0944
Falan Han1, Shuchao Pang2, Yinghua Cui3, Bo Yan2,4,5
1Cheeloo College of Medicine, Shandong University, Jinan 250012, China
2Shandong Provincial Key Laboratory of Cardiac Disease Diagnosis and Treatment, Affiliated Hospital of Jining Medical University, Jining Medical University, Jining 272029, China
3Division of Cardiology, Affiliated Hospital of Jining Medical University, Jining Medical University, Jining 272029, China
4The Center for Molecular Genetics of Cardiovascular Diseases, Affiliated Hospital of Jining Medical University, Jining Medical University, Jining 272029, China
5Shandong Provincial Sino-US Cooperation Research Center for Translational Medicine, Affiliated Hospital of Jining Medical University, Jining Medical University, Jining 272029, China
OBJECTIVES Acute myocardial infarction (AMI), a common complex disease caused by an interaction between genetic and environmental factors, is a serious type of coronary artery disease and is also a leading cause of death worldwide. Autophagy-related 16-like 1 (ATG16L1) is a key regulatory factor of autophagy and plays an important role in induced autophagy. In the cardiovascular system, autophagy is essential to preserve the homeostasis and function of the heart and blood vessels. Massive gene sequencing effort and function research promote a better understanding of causal risk factors. There are no studies on the association between ATG16L1 and AMI. To identify genetic risk loci for acute myocardial infarction and determine their functional role in disease development, we conducted a case-control study of ATG16L1 gene promoter versus acute myocardial infarction.
METHODS We conducted a case-control study, using polymerase chain reaction and sequencing techniques, dual-luciferase reporter assay, and electrophoretic mobility shift assay to analyze genetic and functional variation in the ATG16L1 gene promoter between AMI and controls. A variety of statistical analyses were used to analyze the allele and genotype frequencies and the relationship between DSVs and AMI. The TRANSFAC database was used to predict the relevant transcription factors for ATG16L1 gene promoter polymorphism. Values of P<0.05 were considered to indicate statistical significance.
RESULTS In all, ten SNPs and two DNA-sequence variants (DSVs) were identified in 688 subjects, and three ATG16L1 gene promoter mutations (g.233250693 T>C [rs185213911], g.233250946 G>A [rs568956599], g.233251133 C>G [rs1301744254]) that were identified in AMI patients significantly altered the transcriptional activity of ATG16L1 gene promoter in HEH2, HEK-293, and H9c2 cells (P<0.05). Further electrophoretic mobility shift assays indicated that the SNPs affected the binding of transcription factors. And by the TRANSFAC database, we found that the SNPs identified in AMI patients can create, modify, and eliminate putative binding sites for transcription factors.
CONCLUSIONS This was the first study to find that the ATG16L1 gene promoter polymorphism is associated with AMI. ATG16L1 gene promoter mutations in AMI patients may affect the binding of transcription factors and change the transcriptional activity of the ATG16L1 gene, changing the level of autophagy and contributing to the occurrence and development of AMI as rare and low-frequency risk factors. This research provides important evidence and insights for molecular studies of cardiovascular diseases. With the development of high-throughput sequencing technology, identifying individuals at presymptomatic genetic risk, evaluating and treating them hold the promise of preventing or improving cardiovascular disease morbidity and mortality. This will also drive the future development of genetic testing to identify subsets of patients at high risk of cardiovascular disease and to adopt the most effective treatment or preventive measures for them, thus achieving precision medicine.
GW31-e0990
Ning Zhang, Xiaojie Xie
Second Affiliated Hospital Zhejiang University School of Medicine
OBJECTIVES Nearly half of all heart failure (HF) patients suffer from Heart failure with preserved ejection fraction (HFpEF), carrying a dismal prognosis without effective targeted therapies. However, the precise pathogenesis responsible for the HFpEF remains unknown. Studies have suggested that HFpEF correlates with macrophage activation and excessive secretion of pro-inflammatory cytokines, which drive left ventricular remodeling and diastolic dysfunction by stiffer cardiomyocytes and interstitial fibrosis. Furthermore, the chemokine CXCL12 and its receptor CXCR4 have been confirmed playing an important role in promoting the infiltration of macrophages into damaged tissues, and then activating the inflammatory cascade. However, whether CXCR4 in macrophages promotes fibrosis in HFpEF is uncleared.
METHODS SAUNA (unilateral nephrectomy and a continuous infusion of d-aldosterone (0.30 μg/h) via osmotic minipumps (Alzet) and salty (1% NaCl) drinking water) was administrated to wild-type (WT) mice and myeloid-specific CXCR4-deficient mice or bone marrow (BM) reconstituted chimeric mice for 30 days. In vitro, we first detected macrophage migration and cytokines secretion after HMGB1 treatment and explored the role of CXCR4 and Lyn interaction in macrophages. Then, we evaluated the paracrine effect from activated macrophages on fibroblasts through co-cultured with macrophage and fibroblasts.
RESULTS Loss of the myeloid cells-expressed CXCR4 in mice (MKO) markedly reduced cardiac inflammation, hypertrophy, fibrosis and diastolic dysfunction of HFpEF. These alterations were macrophage dependent because MKO compared with WT mice show decreased infiltration of macrophages but not neutrophils into the hearts at 30 days after SAUNA, whereas bone marrow transplantation from WT mice into MKO mice rescued the diastolic dysfunction and vice versa. Consistent results were seen in vivo, suppressed Lyn phosphorylation contributed to impaired migration and cytokine secretion in CXCR4 KO macrophages. Additionally, primary fibroblasts co-cultured with CXCR4 knockout macrophages showed repressed differentiation of cardiac fibroblasts into myofibroblasts and blocked the synthesis of extracellular matrix (ECM), confirming that macrophages influence cardiac fibrosis in a CXCR4-dependent paracrine manner through TGFβ-smad2/3 signal pathway.
CONCLUSIONS Our findings elucidate that CXCR4 in macrophages can affect left ventricular diastolic dysfunction by increasing macrophage infiltration and enhancing inflammatory secretion, and resulting in inflammation response and interstitial fibrosis.
GW31-e1033
Dongfei Wang, Xiaogang Guo
Department of Cardiology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
OBJECTIVES Dilated cardiomyopathy (DCM) is considered as the most common form of non-ischemic cardiomyopathy with a high mortality worldwide. Cytoskeleton protein Cypher plays an important role in maintaining cardiac function. Genetic studies in human and animal models revealed that Cypher is involved in the development of DCM. However, the underlying molecular mechanism is not fully understood. Accumulating evidences suggest that apoptosis in myocytes may contribute to DCM. Thus, the purpose of this study is to define whether lack of Cypher in cardiomyocytes can elevate apoptosis signaling and lead to DCM eventually.
METHODS We used siRNA to downregulate the expression of Cypher in H9c2 cells and neonatal rat cardiomyocytes which was confirmed by RT-qPCR and western blot. Cell counting kit-8 assay was used to value cell viability. TUNEL and Flow cytometry were conducted to confirm the apoptosis of the cardiomyocytes. RT-qPCR and western blot were further used to detect the specific signal pathway contributed to the apoptosis of the cardiomyocytes.
RESULTS Cypher-siRNA sufficiently inhibited Cypher expression in cardiomyocytes. TUNEL-positive cardiomyocytes were increased in both Cypher knockdown neonatal rat cardiomyocytes and Cypher knockout mice hearts, which were rare in the control group. Flow cytometry further confirmed that downregulation of Cypher significantly increased myocytes apoptosis in vitro. Cell counting kit-8 assay revealed that Cypher knockdown in H9c2 cells significantly reduced cell viability. Cypher knockdown was found to increase cleaved caspase-3 expression and suppress p21, ratio of bcl-2 to Bax. Cypher-deficiency induced apoptosis was linked to downregulation of Akt activation and elevated p-p38 MAPK accumulation. Pharmacological activation of Akt with SC79 attenuated apoptosis with enhanced phosphorylation of Akt and reduced p-p38 MAPK and Bax expression.
CONCLUSIONS Downregulation of Cypher participates in the promotion of cardiomyocytes apoptosis through inhibiting Akt dependent pathway and enhancing p38 MAPK phosphorylation. These findings may provide a new potential therapeutic strategy for the treatment of DCM.
GW31-e1035
Shiyu Zhang, Youping Wang
Central Laboratory and Division of Cardiology, First Affiliated Hospital, Henan University of Traditional Chinese Medicine
OBJECTIVES Our prior studies show that transient receptor potential vanilloid type 1 (TRPV1) exerts protective effects on renal inflammatory injury in salt-sensitive hypertension. However, the mechanisms are not determined. It is well accepted that endothelial dysfunction, especially a reduction of PI3K/Akt-dependent endothelial nitric oxide synthase (eNOS)-induced nitric oxide (NO) production, contributes to the development of inflammation. Recently, compelling evidence shows that TRPV1, originally cloned from primary sensory nerves, is also expressed in endothelial cells and is involved in regulating endothelial cell function. Thus, we aimed to test the hypothesis that NO derived from Ca2+/PI3K/Akt-dependent eNOS contributes to TRPV1-induced anti-inflammatory effects in vitro in human umbilical vein endothelial cells (HUVECs).
METHODS Human umbilical vein endothelial cells (HUVECs) were cultured in the presence of capsaicin (CAP, a specific TRPV1 agonist) with or without the specific antagonist of TRPV1, NOS, or Ca2+/PI3K/Akt pathway prior to lipopolysaccharide (LPS) stimulation. NO metabolites, protein expression, and inflammatory molecules were determined using a Griess assay, Western blot, and immune assay-based multiplex analysis, respectively. Monocyte adhesion was evaluated by assaying the fluorescently labeled human monocytes that adhered to LPS-stimulated endothelial cells.
RESULTS Treatment with CAP (3 or 10 μM) for 6 h dose-dependently increased NO production, and the NO production was time-dependently increased in response to CAP (10 μM) in HUVECs. The CAP-mediated NO production was associated with increased eNOSser1177 phosphorylation. The CAP-induced effects were abolished by pretreatment with capsazepine (CAPZ, a specific TRPV1 antagonist, 10 μM), LNMMA (a non-selective inhibitor of NOS, 1 mM), LY294002 (a specific inhibitor of PI3K/Akt pathway, 10 μM), or EGTA (500 nM) that removes extracellular Ca2+ from the medium for 1 h (P<0.05). In HUVECs, pretreatment with CAP (10 μM) for 1 h inhibited LPS (1 μg/mL)-induced increases in the production of proinflammatory cytokines/chemokines including TNF-α, IL-6 and MCP-1, the expression of adhesion molecules including ICAM-1 and VCAM-1, NF-κB activity, and the number of monocytes that adhered to HUVECs (P<0.05). Moreover, the inhibitory effects induced by CAP were completely abolished by concurrent treatment with CAPZ (10 μM), LNMMA (1 mM), or EGTA (500 nM) (P<0.05).
CONCLUSIONS Our study provides, for the first time, a direct link between TRPV1 and Ca2+/PI3K/Akt/eNOS/NO pathway in TRPV1-mediated anti-inflammatory actions in endothelial cells. Our data indicate TRPV1 activation suppresses endothelial cell-associated inflammatory response via the activation of Ca2+/PI3K/Akt/eNOS/NO pathway. (Corresponding author: Youping Wang. This work was supported by a grant from the National Natural Science Foundation of China (No. 81170243)).
GW31-e1058
Wande Yu, Hang Zhang
Nanjing First Hospital
OBJECTIVES The present study investigated whether berberine affected Trx1/β-catenin expression and/or activity and if it could reduce the development of pulmonary hypertension in the experimental rat model and the proliferation in human PASMCs (HPASMCs).
METHODS Immunofluorescence and Western blot used to detect Trx1 and β-catenin expression in human pulmonary tissues. All rats were randomized divided into four groups: the control group (n=6), the SU5416/hypoxia (Su/Hox) group (n=7), the Su/Hox+PX12 group (n=7) and the Su/Hox+berbrine group (n=7). In the latter three groups, these rats received injection of SU5416 under isoflurane anesthesia at day 1 and then were exposure to hypoxia started from day 2 to day 29, the third and fourth group also received PX-12 and berberine respectively for 4 weeks. Evaluation of cardiac function using echocardiography. The mean pulmonary artery pressure (mPAP) were measured by right heart catheterization according to Wanghong. Wall thickness and the percentage of medial wall thickness were used to assess the PA remodeling.
RESULTS The results showed that increased proliferation in hypoxia-induced healthy HPASMCs or PAH HPASMCs is associated with a significant increase in Trx1 and β-catenin expression. Treatment with Trx1 specific inhibitor PX-12 significantly reduced pulmonary arterial pressure and vascular remodeling, as well as improved in vivo cardiac function and right ventricular hypertrophy in Su/Hox-induced PAH rats. Berberine reversed right ventricular systolic pressure, right ventricular hypertrophy, and decreased pulmonary vascular remodeling in the rats. Furthermore, berberine has an antiproliferative effect on hypoxia-induced HPASMCs proliferation in a manner likely to be mediated by inhibiting of Trx1 and its target gene β-catenin expression.
CONCLUSIONS Our results illustrate that the expression of Trx1 and beta-catenin contribute to pathological PASMCs proliferation. BBR or PX-12 is a promising therapeutic utility and raise the possibility to alleviate hyperplastic proliferation of HPASMCs and improve cardiac function through manipulation of the expression of the Trx1 and β-catenin.
GW31-e1076
Huishou Zhao, Ling Tao
Department of Cardiology, Xijing Hospital, Air Force Medical University, Xi’an, Shaanxi, China
OBJECTIVES Branched chain amino acids (BCAAs) are associated with the progression of obesity-related metabolic disorders, including type 2 diabetes and nonalcoholic fatty liver disease. However, whether BCAAs disrupt the homeostasis of hepatic glucose and lipid metabolism and contribute to insulin resistance remains unknown.
METHODS we fed mice the ND diet, HF diet, or HF/BCAA diet for 16 weeks. Metabolic studies such as glucose tolerance testes, insulin tolerance tests, pyruvate tolerance tests were determined to examine the insulin resistance. Glucose and lipid metabolic related genes were tested to evaluated the hepatic metabolic homeostasis.
RESULTS In this study, we observed that BCAAs supplementation significantly reduced high-fat (HF) diet-induced hepatic lipid accumulation while increasing the plasma lipid levels and promoting muscular and renal lipid accumulation. Further studies demonstrated that BCAAs supplementation significantly increased hepatic gluconeogenesis and suppressed hepatic lipogenesis in HF-DIO mice. These phenotypes resulted from severe attenuation of Akt2 signaling via mTORC1- and mTORC2-dependent pathways. BCAAs/branched-chain a-keto acids (BCKAs) chronically suppressed Akt2 activation through mTORC1 and mTORC2 signaling and promoted Akt2 ubiquitin-proteasome-dependent degradation through the mTORC2 pathway. Moreover, the E3 ligase Mul1 played an essential role in BCAAs/BCKAs-mTORC2-induced Akt2 ubiquitin-dependent degradation. We also demonstrated that BCAAs inhibited hepatic lipogenesis by blocking Akt2/SREBP1/INSIG2a signaling and increased hepatic glycogenesis by regulating Akt2/Foxo1 signaling. Collectively, these data demonstrate that in DIO mice, BCAAs supplementation resulted in serious hepatic metabolic disorder and severe liver insulin resistance: insulin failed to not only suppress gluconeogenesis but also activate lipogenesis.
CONCLUSIONS Intervening BCAA metabolism is a potential therapeutic target for severe insulin-resistant disease.
GW31-e1077
Huishou Zhao, Ling Tao
Department of Cardiology, Xijing Hospital, Air Force Medical University, Xi’an, Shaanxi, China
OBJECTIVES Diabetic nephropathy (DN) is a major cause of end-stage renal disease (ESRD) with few effective therapeutic strategies. There is an urgent need for new, curative treatments as well as for biomarkers to evaluate risk of DN. Podocyte dysfunction and loss is recognized to be a critical event associated with deterioration of renal function in DN. However, the underlying mechanisms remained elusive. Plasma branched chain amino acids (BCAAs) level is ascended in diabetic patients and is causatively correlated with the development of diabetes. Whether BCAAs accelerate the progression of DN remained unclear.
METHODS We fed mice the ND diet, HF diet, or HF/BCAA diet for 16 weeks. Four percent amino acids mixers (all 20 naturally occurring amino acids except Gln and Asn) were added into drinking water of HF and HF/Paired group. the blood urea nitrogen level, plasma creatinine levels and the ratio of urinary albumin/creatinine were determined to evaluate the renal function; Periodic acid Schiff (PAS) staining, Transmission electron microscope (TEM) analysis were utilized to analyze the glomerular abnormalities. Biochemical methods such as Western blotting and real-time PCR were used to analyze the molecular mechanism.
RESULTS In this study, we observed that BCAAs supplementation exacerbated high fat diet (HF)-induced renal lipid accumulation, proteinuria, glomerulosclerosis, podocyte dedifferentiation, cellular skeletal disordered rearrangement and podocyte loss. Mechanistically, we demonstrated BCAA supplementation induced podocyte metabolic remodeling: enhanced glycolysis, inhibited sugar aerobic oxidation and accelerated serine-one carbon metabolism through depolymerization of pyruvate kinase type M2 (PKM2). BCAAs promoted a shift of PKM2 from tetramers to dimers and monomers and promoted protein and nucleotides synthesis, which elicited endoplasmic reticulum stress (ESR) and led to podocyte loss. PKM2 agonist TEPP46 reversed BCAAs-induced glucose and serine-one carbon metabolic disorders, improved ESR response in podocyte, and ameliorated BCAAs supplementation-induced renal lipid accumulation, proteinuria, and podocytes dysfunctioning and loss. In addition, we demonstrated BCAAs promoted PKM2 expression through activation of mTORC1 signaling, rapamycin treatment eliminated BCAAs supplementation induced up-regulation of PKM2 and produced renal protective effects upon BCAAs induced diabetic kidney injury.
CONCLUSIONS Collectively, we demonstrated BCAAs accumulation accelerated HF-induced renal injury through disrupting podocyte energetic and substance biosynthesis metabolisms, intervening BCAA metabolism or PKM2 may be effective for preventing obesity-associated DN progress.
GW31-e1101
Yujuan Jiang, Xiuyue Jia, Minghong Wang, Xu Han, Hongjuan Xia, Houliang Chen, Chong Li, Xue Zhao
Cardiovascular Department, Eastern Hepatobiliary Surgery Hospital, Shanghai, China
OBJECTIVES The stent therapy of peripheral artery stenosis remains challenging due to high incidence of restenosiscaused by permanent metallic implantation. Metallic stents may act as chronic stimulus for prolonged duration of proliferative response even when the stent surface is completely endothelialized. Besides drug coating balloon and bioresorbable stent, retrievable stent may be another strategy to achieve “nothing left” intervention. This study aimed to explore preliminary the feasibility and the controversial issues of retrievable stent in a canine intact iliac artery.
METHODS The stents were self-developed with counterpoint overlapped double-layer self-expandable drug-eluting scaffolds, which were composed of two-layer scaffolds, inner and outer. A small hook in the outer scaffold was used for retrieval by snare catheter. Four retrievable stents were implanted into the iliac arteries of four Labrador dogs at the same time and retrieved on day 21, 28, 35 and 42 after implantation respectively. The angiography and IVUS were performed before and after stent retrieval. The target vascular segments were harvested for staining of HE, Methyl violet, Masson trichrome and CD31. The dog with stent implanted for 28 days was followed for another 14 days after stent retrieval.
RESULTS The angiography showed all the stents were patency before retrieving. The stents implanted for 28 days (28-day-stent) and 42 days (42-day-stent) were successfully retrieved (Figure 1). IVUS showed a smooth vascular wall before and after stent retrieval. The inner surface of the retrieved 28-day-stent was covered with a white translucent thin layer substance, which was composed of more fibrin and scattered fibroblasts and smooth muscle cells (Figure 2). At the following up of 14 days after stent retrieval, angiography showed the vessel was patency. The artery diameter with 28-day-stent decreased by 10% before retrieval and 21% on day 14 after retrieval compared to baseline. The vascular inner surface was covered with an uneven neointima, which was thicker close to struts and thinner far to struts (Figure 3). The average neointima thickness was 36±2 μm when the gap distance between adjacent struts was ≥200 μm, and it turned to be 163±20 μm when this gap distance was <200 μm. The internal elastic lamina showed breaks in some areas. For the 42-day-stent, some petechiae were observed on the proximal and distal segments in the target artery. Under the stent struts, endothelial cells and smooth muscle cells decreased with local fibrosis, and also the internal elastic lamina was interrupted in partial area (Figure 4). The inner surface of the retrieved stent was covered with a white translucent layer substance, which contained more cell components than that in 28-day-stent, including endothelium and angiogenesis (Figure 2). The media and adventitia were intact. Unfortunately, stents after implantation of 21 days and 35 days failed to be retrieved because of catching failure due to stent displacement.
CONCLUSIONS The study demonstrated the feasibility of retrieving a stent from iliac artery as long as 42 days after implantation. Pulling slightly and slowly or even stopping temporarily was the critical skill to guarantee the higher safety and efficiency of the stent retrieval procedures. A modified stent without hook was under exploration for the further enhancement of retrieval safety. However, neointima formation would still be challenging in further study.
GW31-e1117
Lei Huang1, Yue Zhao1, Lingfang Zeng2, Tong Li1
1Tianjin Third Central Hospital
2School of Cardiovascular Medicine and Sciences, Faculty of Life Science and Medicine, King’s College London
OBJECTIVES Recently, a novel nested intronic gene was discovered from the microarray profiling of the laminar flow-upregulated genes in mouse embryonic stem cells (ESCs). This gene is located in the intron 6 of the lymphoid transcription factor gene Laf4/Aff3. Therefore this novel gene is referred as Laf4 intron resident (Laf4ir). Laf4ir and Laf4 are transcribed using opposite strands of DNA. Laf4ir exhibits 7 exons and two transcript variants (Genbank accession: MH282850.1, GI: 1567433536 and MH282851.1, GI: 1567433538). Laf4ir-tv2 appears to be the dominant transcript variant. Laf4ir has been shown to be a polycistronic gene, encoding a 45-amino acid (aa) peptide from open reading frame 1 (ORF1) and a 109aa (Laf4ir-tv1) or 151aa (Laf4ir-tv2) protein from ORF2, respectively. ORF1 and ORF2 polypeptide expression was found in various adult organs, different stages of embryonic development, different cell types and different subcellular localisation. In this study, we aimed to investigated the potential function of Laf4ir in cardiovascular remodeling and utilized the global knockout mouse model to further understand the underlying mechanisms.
METHODS Mouse ESCs cultured in differentiation medium were subjected to laminar shear at 12 dynes/cm2 for 24 h. The 3-day spontaneously differentiated ESCs were infected with Ad-null or Ad-L151 (the adenovirus contains the cDNA sequence for the 151-aa of ORF2) at 10MOI for 16 h. The cells were then seeded and incubated for 24 h followed by the Br-dU incorporation assay with the cell proliferation. Mouse ECs C166 were seeded in complete growth medium for sixteen hours, then adenoviral particles were added at 10 MOI and incubated for 24 h. The cells were eventually incubated with serum free medium containing H2O2 at concentration indicated for 24 h. MTS assay and Annexin V/Dead Cell Apoptosis assay were performed accordingly to assess the cell survival. To investigate the potential contribution of Laf4ir in cardiovascular repair and pathology, transverse aortic constriction-mediated pressure overload in heart, aorta from ApoE-/- mice, femoral artery wall following vascular injury and ischemia were investigated. To explore the functionality of Laf4ir further, a Cre-loxP in-vivo global knockout model was developed and utilized.
RESULTS Overexpression of LAF4IR-ORF2 via adenoviral gene transfer could enhance laminar flow- and VEGF-induced endothelial cell (EC) differentiation and reduce cell proliferation. This could be due to cell cycle arrest via the retention of mini-chromosome maintenance protein 3 (MCM3) in the cytosol and as a consequence assist differentiation towards EC lineage. Overexpression of LAF4IR-ORF2 also promoted endothelial survival under oxidative stress induced by hydrogen peroxide. The upregulation of ORF2 polypeptide was detected in transverse aortic constriction-mediated pressure overload in heart, aorta from ApoE-/- mice, femoral artery wall following vascular injury and ischemia. Vascular ischemia in Laf4ir knockout mice confirmed the contribution of Laf4ir in vascular repair. Sca1+ adventitia cells isolation from transgenic mice further validated functions of Laf4ir in proliferation.
CONCLUSIONS Overall, the novel nested Laf4ir gene may contribute to cardiovascular remodeling through spatiotemporal translation of different ORFs. Further detailed investigation on Laf4ir will undoubtedly yield new insights into cardiovascular physiology and pathology.
GW31-e1123
Sanjiu Yu, Jihang Zhang, Lan Huang
Institute of Cardiovascular Diseases, Xinqiao Hospital, Army Medical University
OBJECTIVES Oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell (EC) apoptosis is the initial step of atherogenesis. Mitochondria-associated endoplasmic reticulum (ER) membrane (MAM), regulated by tethering proteins such as phosphofurin acidic cluster sorting protein 2 (PACS2), is essential for mitochondrial Ca2+ overload and subsequent cell apoptosis. Previously, we demonstrated that PACS2 played an important role in ox-LDL-induced EC apoptosis by regulating MAM formation, but the underlying mechanism was still unclear. In this study, we aimed to investigate the upstream of PACS2 in ox-LDL-induced apoptosis in human umbilical vein endothelial cells (HUVECs).
METHODS HUVECs were treated with ox-LDL at 0, 50, 100, 150, 200 or 250 μg/mL for 24 h or at 200 μg/mL for 0, 6, 12, 24, 48 or 72 h. Mitochondrial Ca2+ uniporter (MCU) and PACS2 were knocked down by using small interfering RNA. After treatments, cell viability and apoptosis were measured by cell counting kit-8 assay and Annexin V/propidium iodide staining, respectively. Mitochondrial membrane potential (MMP) was evaluated by JC-1. Reactive oxygen species (ROS) was examined by DCFH-DA. Mitochondrial Ca2+ level was assessed by fluorescent probe Rhod-2 AM. The mRNA or protein levels of target genes were determined by quantitative real-time polymerase chain reaction or western blotting. MAM formation was detected by confocal microscopy and transmission electron microscopy.
RESULTS Ox-LDL dose- and time-dependently increased cell apoptosis concomitant with mitochondrial Ca2+ elevation, mitochondrial membrane potential (MMP) loss, reactive oxygen species (ROS) production. Besides, ox-LDL upregulated PACS2 expression and decreased miR499 level. Further, we found that over-expression of miR-499 could protect HUVECs against ox-LDL-induced apoptosis. Increased miR-499 level favored endothelial cell survival, while decreased miR-499 level favored apoptosis. In addition, over-expression of miR-499 could protect HUVECs against ox-LDL-induced mitochondrial Ca2+ elevation, MMP loss, ROS production. Further, we identified PACS2 as the target of miR-499. MiR-499 inhibited endothelial apoptosis through its suppressive effect on PACS2 expression, thereby blocking ox-LDL-induced MAM formation and ER-mitochondria Ca2+ transfer.
CONCLUSIONS Altogether, our findings suggest that miR-499 plays an inhibiting role in ox-LDL-induced EC apoptosis by regulating PACS2 expression, MAM formation and mitochondrial Ca2+ elevation, implicating that miR-499 may be a promising therapeutic target for atherosclerosis.
GW31-e1126
Yan Zhang, Chunyu Zeng
Department of Cardiology, Daping Hospital, The Third Military Medical University
OBJECTIVES Patients with obesity-related hypertension have impaired sodium excretion. However, the mechanisms are incompletely understood. Adipocytes secrete numerous hormones, called adipokines, among which adiponectin is an important one. Whether and how adiponectin contributes to impaired sodium excretion in hypertension has not been previously investigated. The current study tested the hypothesis that adiponectin promotes natriuresis and diuresis and their impairment is involved in hypertension.
METHODS We used Wistar-Kyoto (WKY) rats, spontaneously hypertensive rats (SHRs), GRK4 142A>V transgenic mice to test the natriuresis-effect of adiponectin through intrarenal arterial infusion. Renal proximal tubule (RPT) cells, adiponectin knock out (Adipo-/-) mice, mutant plasmid were used.
RESULTS We demonstrate that sodium excretion was reduced in Adipo-/- mice. The intrarenal arterial infusion of adiponectin induced natriuresis and diuresis in WKY rats, which was impaired in SHRs. Adiponectin inhibited Na+-K+-ATPase activity in RPT cells from WKY rats but not from SHRs. Increased adiponectin receptor phosphorylation and subsequent uncoupling from Gαi, rather than altered adiponectin receptor expression, were responsible for the loss of adiponectin-mediated inhibition of RPT Na+-K+-ATPase activity, impaired natriuresis and diuresis in SHRs. Mutation of the AdipoR1 phosphorylation site restored its linkage to Gαi and the adiponectin-mediated inhibition of Na+-K+-ATPase activity in RPT cells from SHRs. Finally, we identified G protein-coupled receptor kinase 4 (GRK4) as the cause of adiponectin receptor hyper-phosphorylation. GRK4 142A>V transgenic mice replicated the abnormal adiponectin function in SHRs, whereas down-regulation of GRK4 by renal-ultrasound directed siRNA restored the adiponectin-mediated sodium excretion and reduced the blood pressure in SHRs.
CONCLUSIONS Our present study indicates that the stimulatory effect of adiponectin on sodium excretion is impaired in hypertension, which is attributable to increased renal GRK4 expression and subsequent adiponectin receptor hyper-phosphorylation. Targeting GRK4/adiponectin receptor/Gαi may restore the impaired adiponectin-mediated sodium excretion in hypertension, thus representing a novel strategy against hypertension, particularly those patients with obesity-related hypertension.
GW31-e1143
Min Han, Yi Tong Ma
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, P.R. China
OBJECTIVES The rate of aging of the global population is growing, which contributes to increasing incidence and morbidity of acute myocardial infarction (AMI) in worldwide. Although the reperfusion therapy is currently regarded as the best strategy for AMI, the ischemic/reperfusion (I/R) injury leads to a poor prognosis, especially in the elderly patients. Pyroptosis is a pro-inflammatory programmed cell death that is related to myocardial I/R injury, but its effect has not been reported in aged heart. The present study is designed to investigate the underlying mechanism of pyroptosis in senile cardiomyocytes or heart.
METHODS First, we investigated the differences in the infarct size and left ventricular remodeling following I/R surgery between aged (18-mon) and young (3-mon) C57Bl/6 male mice and explored responsible mechanisms. Then, the AAV9 vector carrying mutant IκBαSer32A,Ser36A gene were delivered to senile cardiomyocytes or 18-month-old C57BL/6 mice prior to hypoxia/reoxygenation (H/R) or I/R treatment. Cytosolic IκBα and p65/p50 nuclear translocation were identified by Western blot. The level of the pyroptosis was detected by Western blot and ELISA.
RESULTS The infarct size after I/R was significantly higher in aged than that in young mice (57.89±2.55% vs. 47.03±3.05%, P<0.05). Echocardiography revealed more profound LV chamber dilatation and dysfunction in aged mice. Subsequently, more dramatic IκBα degradation and nuclear translocation of NF-κB p65/p50 were found in aged than in young I/R hearts, in keeping with enhanced oxidative injury. Furthermore, we observed significantly higher contents of serum LDH and IL-1β, and markedly enhanced expression of gasdermin D-N (GSDMD-N) domains in the aged than in young mice. After transfection with the AAV9-IκBα into senescent cardiomyocyte or aged heart, the levels of NLRP3, Caspase-1 and GSDMD-N were downregulated during H/R or I/R treatment. Moreover, IκBα increased cell viability and decreased PI-positive cells in vitro, reduced the myocardial infarct size and the cardiac remodeling in vivo via suppressing NF-κB activation. Additionally, IκBα transfection significantly downregulated the oxidative injury as well as the levels of supernatant or serum LDH and IL-1β in senile cardiomyocyte or aged mice.
CONCLUSIONS This study confirms that aged mice had a worse outcome of I/R injury accompanied with a higher level of NF-κB activation and pyroptosis. Both in vivo and in vitro, IκBα transfection was able to alleviate myocardial I/R injury and inhibit pyroptosis in senile cardiomyocytes or aged heart inhibitory effect of IκBα on pyroptosis was mediated by suppressing the NF-κB/NLRP3/GSDMD axis. Therefore, targeted inhibition of NF-κB signaling may provide an alternative treatment for myocardial I/R injury in aged heart.
GW31-e1155
Jing Ming, Yi Tong Ma
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, P.R. China
OBJECTIVES Dyslipidemia is one of the main risk factors for coronary heart disease (CHD). The E3 ubiquitin ligase which is encoded by the ring finger protein 145 (RNF145) gene is very important in the mediation of cholesterol synthesis and effectively treats hypercholesterolemia. Thus, the purpose of the present research is to investigate the connection between the polymorphism of the RNF145 gene and cholesterol levels in Xinjiang, China
METHODS A total of 1396 participants (male: 628, female: 768) were included in this study for genetic analysis of RNF145 gene, and we used the modified multiple connection detection response (iMLDR) technology to label two SNPs (rs17056583, rs12188266) of RNF145 genotyping. The relationship between the genotypes and the lipid profile was analyzed with general linear model analysis after adjusting confounding variables.
RESULTS Through the analysis of the two SNPs in RNF145 gene, we discovered that both rs17056583 and rs12188266 were related to total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) concentrations (All P<0.001). Compared to C, the G allele of rs17056583 has significantly lower plasma TC concentrations (2.76±0.70 mmol/L, 6.29±1.04 mmol/L, P<0.001) and LDL-C concentrations (1.27±0.29 mmol/L, 5.10±0.60 mmol/L, P<0.001). The G allele carrying rs12188266 had significantly higher TC concentrations (6.39±0.98 mmol/L (2.95±0.99 mmol/L, P<0.001) and LDL-C concentrations (5.15±0.55 mmol/L, 1.44±0.70 mmol/L, P<0.001) than those carrying the A allele. Meanwhile, we also found that rs17056583 was associated with high triglycerides concentrations (All P<0.001). In addition, the association of rs17056583 and rs12188266 with lipid profile concentrations is still statistically significant after multivariate adjustment of sex, age, smoking, obesity, alcohol consumption, diabetes, and hypertension.
CONCLUSIONS Our study shows that both rs17056583 and rs12188266 SNPs of RNP145 gene are related to TC and LDL-C concentrations in Xinjiang population.
GW31-e1160
Hui Wang, Yi Zhu
Tianjin Key Laboratory of Metabolic Diseases; Key Laboratory of Immune Microenvironment and Disease (Ministry of Education); Collaborative Innovation Center of Tianjin for Medical Epigenetics and Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China
OBJECTIVES Vascular endothelial cells (ECs) are of critical importance for maintaining blood pressure (BP). Although cartilage oligomeric matrix protein (COMP) plays a pivotal role in maintaining cardiovascular homeostasis, its effect on endothelial function and blood pressure is largely unknown. In this study, we investigated the protective effect of COMP on blood pressure and its underlying mechanism.
METHODS Wild type (WT) mice and COMP-/- mice were infused with angiotensin II (450 ng/kg/min) for 3 days via an osmotic minipump, and BP was monitored by a tail-cuff system. Second-order mesenteric arteries were isolated from mice for microvascular tension measurement in an oxygenated organ chamber. Nitric oxide (NO) was detected by an electron paramagnetic resonance (EPR) technique using iron (II) diethyldithiocarbamate [Fe (DETC)2] as the spin trap. To determine intracellular calcium influx, endothelial cells were loaded with a fluorescent probe fluo-4AM, and live cell images were acquired with a confocal microscope. Small interfering (si) RNA transfection, western blot, and co-immunoprecipitation were used for further detailed mechanism investigation.
RESULTS COMP deficiency showed higher blood pressure than WT mice in baseline or after angiotensin II infusion. Disruption of COMP impaired acetylcholine-evoked endothelium-dependent relaxation (EDR) in second-order mesenteric arteries as compared with WT mice. L-NG-nitroarginine methyl ester (L-NAME), an endothelial nitric oxide synthase (eNOS) inhibitor, abolished the difference in EDR between the two genotypes. Mechanistically, COMP increased intracellular calcium influx, which promoting the dissociation of eNOS and caveolin-1, augmenting the phosphorylation of Ca2+/calmodulin-dependent protein kinases II (CaMKII) and eNOS. Furthermore, co-immunoprecipitation assays suggested that COMP bound directly to the endothelial mechanosensitive cation channel PIEZO1. SiRNA-mediated PIEZO1 deficiency blocked COMP-dependent eNOS activation. Meanwhile, PIEZO1 activator Yoda1 reduced the difference of EDR in WT and COMP-/- mice.
CONCLUSIONS Our study demonstrates that COMP plays a critical role in blood pressure homeostasis by interacting with PIEZO1, and increasing intracellular calcium influx, eNOS activity and NO formation.
GW31-e1165
Sai Ma1,2, Ren Jun2
1Jinling Hospital, Medical School of Nanjing University
2University of Wyoming
OBJECTIVES Nicotinamide riboside (NR) is widely used as a NAD+ precursor vitamin. Supplementation with NR has been shown to protect against metabolic disease in mammals. However, the potential effect of NR in alcoholic cardiomyopathy (ACM) has not been well elucidated. This study was designed to examine the effect of NR supplementation on the progression of alcoholic cardiomyopathy.
METHODS Alcoholic cardiomyopathy was established using chronic alcoholic diet containing 36% kcal from ethanol. Echocardiography and IonOptixMyoCam were used to evaluate cardiac contractile function.
RESULTS Our data revealed that NR alleviated alcohol consumption-induced changes in myocardial and cardiomyocyte contractile function as well as cardiac remodeling. To examine the possible involvement of mitophagy in NR-induced beneficial effects, FUNDC1-/- mice with mitophagy deficiency were employed. Interestingly, NR-induced beneficial effect against alcoholic cardiomyopathy was partially attenuated in FUNDC1-/- mice, indicating a role for FUNDC1-mediated mitophagy in NR-offered cardioprotection. In vitro study using H9c2 myoblasts suggested that NR regulated mitochondrial homeostasis through induction of FUNDC1-dependent mitophagy, as suggested by mitophagy-related protein expressions and Mitotracker-LC3 dots overlay. NR treatment enhanced cellular NAD+ level, consequently elevated NAD+-dependent mitochondrial sirtuin SIRT3 activity. Using mass spectrum assay and Co-IP, PGAM5, which functions to phosphorylate FUNDC1 at serine 13 (Ser13), was found to interact with and deacetylated by SIRT3 following NR administration.
CONCLUSIONS Taken together, our results revealed a protective effect of NR supplementation against alcoholic cardiomyopathy possibly associated with a SIRT3-PGAM5-FUNDC1-dependent regulation of mitophagy. These findings suggested the therapeutic potential of the vitamin B3 precursor of NAD+ in the management of alcoholic cardiomyopathy.
GW31-e1173
Jing Cao1, Zhaoya Liu2, Qian Xu3, Guogang Zhang1, Ruizheng Shi4
1Department of Cardiovascular Medicine, The Third Xiangya Hospital, Central South University, 410013 Changsha, China
2Department of Geriatrics, The Third Xiangya Hospital, Central South University, 410013 Changsha, China
3Department of Cardiothoracic Surgery, Xiangya Hospital, Central South University, 410008 Changsha, China
4Department of Cardiovascular Medicine, Xiangya Hospital, Central South University, 410008 Changsha, China
OBJECTIVES It is well known that necrosis due to coronary artery occlusion is the main form of cardiomyocytes loss. Recently, there are a considerable number of studies focusing on non-necrotic cell death of cardiomyocytes stimulated by hypoxia and inflammatory factors in infarction border zone, which contributes a lot to the loss of interventional cells. Therefore, it is urgent to explore the regulatory mechanism of non-necrotizing myocardial cell death to reduce the loss of cardiomyocytes after myocardial infarction (MI). Ferroptosis that depends on iron and reactive oxygen species is a newly discovered way of programmed cell death. Myocardial iron level has been addressed as an important prognostic factor for heart failure after MI in numerous clinical studies. However, the mechanism of ferroptosis in cardiomyocytes after MI has not been fully elucidated. Peroxidasin (PXDN) is a member of the peroxidase family which contain heme groups in its structure domain and can generate hypochlorous acid (HOCl) by consuming hydrogen peroxide (H2O2). Previous studies have found that PXDN level is significantly increased in the plasma of patients with MI, but the specific mechanism of PXDN in MI remains unclear.
METHODS Eight-week-old systemic knockout of PXDN (PXDN-/-) and homologous control mice (PXDN+/+) were used to construct a model of myocardial infarction with left coronary artery ligation. Echocardiography and three-dimensional speckle tracking imaging were applied to assess left ventricular function and fibrosis. Effects of PXDN changes on infarction area, myocardial cell death and degree of myocardial fibrosis after MI were evaluated by Cardiac TTC, live/dead and masson staining. The expression and localization of non-heme, PXDN, heme oxygenase-1 (HO-1), 3-chlorot-yrosine (3-Cl-Tyr) in cells were detected by immunohistochemistry, iron staining, and immunofluorescence. Levels of PXDN, 3-Cl-Tyr, HO-1, ferroptosis-related marker and lipid peroxidation products were confirmed by Real-time PCR and western blot. In vitro experiments were performed in H9C2 cells. H9C2 cells with PXDN knockdown by siRNA were pretreated with 1% O2 or ferric citrate. Cell death, expression of PXDN, 3-Cl-Tyr, HO-1, ferroptosis biomarker and lipid peroxidation products were detected by immunofluorescence, western blot and real-time PCR.
RESULTS Mice after MI had significantly weakened cardiac function, and severe myocardial fibrosis in the infarct area, accompanied with increased ferroptosis. Expression of PXDN, HO-1 and 3-Cl-Tyr were increased in mice myocardium after MI, whereas TfR were decreased. The cardiac iron levels, lipid peroxidation and ferroptosis of cardiomyocytes were decreased in Mice lacking PXDN, resulting in more mild cardiac function injury and fibrosis upon MI and increased survival rate. At the molecular level, mice with PXDN systemic knockout had lower level of non-hem, 3-Cl-Tyr and ferroptosis-related marker and higher expression of TfR. Expression of PXDN, HO-1 and 3-Cl-tyr in H9C2 cells were elevated during hypoxic conditions, as well as the biomarker of ferroptosis (ptgs2, NOX1 and COX2), lipid peroxidation level and cell death. Moreover, intracellular iron levels were increase together with expression of TfR in cell membrane decreased. These results had been found in cells treated with exogenous iron (ferric citrate) and could be reversed after PXDN silence.
CONCLUSIONS Lacking of PXDN in mice reduces the occurrence of myocardial ferroptosis after MI and improves the prognosis of MI.
GW31-e1206
Dan Zhu, Wei Chen, Xinyang Hu, Jianan Wang
Second Affiliated Hospital of Zhejiang University School of Medicine
OBJECTIVES Mesenchymal stem cell (MSC) injection after myocardial infarction promotes recovery of heart function. But how matrix deposition and collagen crosslinking during the healing process affects cardiac stiffness and MSC engraftment are unknown. Cell probes rigidity by applying forces to the ECM through actomyosin-focal adhesions connections. However, whether rigidity influences contractile machinery assembly after initial contact with ECM during cell spreading is unknown. Focal adhesion forms under lamella network after lamellipodia retraction. Cofilin is an actin binding protein best known for its role in actin filament severing and lamellipodia network turnover. Here we investigate the role of cofilin in contractile machinery assembly, rigidity sensing and MSC engraftment.
METHODS Cardiac stiffness was measured by atom force microscopy. MSC engraftment was detected by immunofluorescence staining and PCR. Cell tractions exerted on substrate of different rigidity was detected by traction force microscopy. Actin retrograde flow (ARF) was measured by time-lapse microscopy. Cofilin activation after PIP2 hydrolysis was visualized by fluorescence resonance energy transfer (FRET) microscopy. Integrin–fibronectin interaction was measured by biomembrane force probe (BFP).
RESULTS Cardiac stiffness increased over time after myocardial infarction. MSC engraftment and adhesion increased with substrate stiffness. ARF, traction force and cofilin activity increased with substrate stiffness. PIP2 and cofilin association decreased on stiffer substrate. Tropomyosin enriched in lamellipodia on stiffer substrate. Constitutive activated cofilin promotes ARF, tropmyosin enrichment and traction force on soft substrate, whereas branch dissociation defects decreased ARF, tropmyosin enrichment and traction force on stiffer substrate. Increasing force loading rate prolonged integrin and fibronectin bond lifetime.
CONCLUSIONS Our data demonstrate that increasing substrate rigidity depletes PIP2 level on cell membrane, which leads to cofilin translocation and activation. Cofilin activation induces lamellipodia actin debranching and contractile machinery assembly. Contractile machinery assembled on stiff substrate promotes ARF and force loading rate on integrin–ECM bond, which underlies enhanced MSC adhesion and engraftment after myocardial infarction.
GW31-e1209
Ling Zhang, Kaiqi Lv, Xinyang Hu, Jianan Wang
Second Affiliated Hospital Zhejiang University School of Medicine
OBJECTIVES Cell therapy showed great potentials in the regenerative and related medicine in the past ten years, however, low survival rates and short residence time of transplanted cells staying at disease region after their implantation have hampered their functional outcomes. The purpose of the present study was to investigate whether single cell engineering with surface-conjugated gelatin could improve mechenchymal stem cell survival and therapeutic effects for myocardial infarction (MI).
METHODS Here, we artificially synthesize poly (sialic acid) based biocompatible anchor molecule that can self-assemble in the phospholipid bilayer of mesenchymal stem cell (MSC) membrane and which concatenates with microbial transglutaminase that can lead to catalytic formation of gelatin hydrogel on the surface of single MSCs. Then the cell survival after single cell engineering was detected in the hypoxia and ischemia environment in vitro and in vivo, and the cardiac function after cell transplantation was also investigated by echocardiography and Masson’s trichrome staining. Furthermore, the underlying mechanism of cell protection against apoptosis by hydrogel encapsulation was detected by RNA-seq and western blot.
RESULTS In the present study, we report a novel approach to encapsulating a single MSC in a layer of gelatin by surface engineering with encapsulation efficiencies 94%. Our results show that in a single MSC modification significantly increase the ability to resistance to stress while the modification did not affect the paracrine effects of MSC in vitro. Bioluminescence images indicates that hydrogel engraftation improves MSC survival in vivo, moreover, the cell survival was further confirmed by GFP PCR and GFP immunostaining after transplanting GFP transfected MSCs in MI model. We also observed that hydrogel-engrafted MSCs protect cardiac cells against apoptosis and enhance angiogenesis in heart repair, and the MSCs-Gel transplantation results in better cardiac function recovery. Finally, Tnfrsf19/NFκB pathway was found to be involved in the cell protection against apoptosis by hydrogel encapsulation.
CONCLUSIONS The single cell-hydrogel engraftation provides a completely different vision of cell engineering and should find use in a variety of translational applications and hold great potentials for cell therapy of diseases.
GW31-e1211
Liangpeng Li, Chunyu Zeng
Deparment of Cardiology, Army Medical Center (Daping Hospital), Army Medical University, Chinese People’s Liberation Army
OBJECTIVES G protein coupled receptor kinase 4 (GRK4) plays an important role in the development of hypertension, but its effect on cardiac ischemia injury is not clear. The aim of this study is to investigate the effect of GRK4 on myocardial infarction and its underlying mechanism.
METHODS In C57/BL6 mice, we assessed the expression and distribution of GRK4 in the basal state and at different time after MI. GRK4 A486V (GRK4 486 glycine mutation to valine) is a variant of GRK4 in human. Its distribution frequency in Asian people is about 66.4%, and its enzyme activity is higher than that of GRK4 wild type (GRK4 WT). We generated GRK4 WT and GRK4 A486V transgenic mice, as well as cardiac GRK4 knockout mice. We measured the cardiac function, infarct size, cardiomyocyte apoptosis and autophagy after MI, and examined the interaction between GRK4 and HDAC4 and its effect on Beclin1 expression. We also analyzed the correlation between cardiac function and GRK4 A486V variant in 550 patients with AMI.
RESULTS The mRNA and protein levels of GRK4 in the heart of C57/BL6 mice increased significantly 48 hours after MI, and were mainly distributed in the nucleus. Hypoxia induced the increase of GRK4 mRNA and protein levels and nuclear entry of primary cardiomyocytes. Compared with control group, the GRK4 WT transgenic mice exhibited impaired cardiac function of, enlarged infarct size and increased mortality, these were more significant in GRK4 A486V transgenic mice. On the contrary, cardiac GRK4 knockout mice had better cardiac function and smaller infarct size than the control group. TUNEL assays showed that overexpression of GRK4 WT and GRK4 A486V increased cardiomyocyte apoptosis. Overexpression of GRK4 decreased the LC3 II levels and the GFP puncta in primary cardiomyocyte transfected with GFP-LC3 adenovirus. Rapamycin, an autophagy agonist, partially rescued the autophagy inhibition and apoptosis induced by GRK4 overexpression in primary cardiomyocytes. Overexpression of GRK4 reduces the expression of Beclin1, which is positively regulated by histone deacetylase 4 (HDAC4) in the nucleus, while GRK4 reduces the nuclear entry of HDAC4 after hypoxia. HDAC4 S3A (HDAC4 246/467/632 serine to alanine mutation, which leads to phosphorylation resistance) ameliorated inhibition of autophagy and promotion of apoptosis induced by GRK4. The phosphorylation of HDAC4 632 serine was further confirmed by immunoprecipitation and Western blot after myocardial infarction. HDAC4 S632A (serine to alanine mutation at HDAC4 632, resistant to phosphorylation) decreased the inhibition of GRK4 on autophagy. Immunoprecipitation and proximal ligation assay indicated that GRK4 interacted with HDAC4 in mouse heart. Chip assay showed that HDAC4 bound to the promoter region of Beclin1, while GRK4 weakened this binding. The multiple regression analysis of 550 patients diagnosed with acute myocardial infarction showed that, the cardiac function of GRK4 A486V carriers were worse than that of GRK4 WT genotype carriers, GRK4 A486V genotype is an independent factor for predicting lower cardiac function after myocardial infarction.
CONCLUSIONS The expression of GRK4 increases in infarcted mice heart and it translocates into the cardiomyocyte nuclei. GRK4 inhibits autophagy and promotes apoptosis in cardiomyocyte after MI. This effect is depends on the phosphorylation of HDAC4 on 632 serine by GRK4 and the subsequent nuclear-export of HDAC4 which leads to down regulation of Beclin1.
GW31-e1213
Qingju Li, Yinchuan Xu, Kaiqi Lv, Xinyang Hu, Jianan Wang
Second Affiliated Hospital Zhejiang University School of Medicine
OBJECTIVES The potency of mesenchymal stem cells (MSCs) for treatment of myocardial infarction (MI) in nonhuman primates (NHPs) can be significantly improved by culturing the cells under hypoxic conditions before administration, and most of the improvement is likely caused by increases in the cells’ paracrine activity, which are often transported to their target cells by exosomes. Thus, exosomes have a key role in mechanisms that regulate apoptosis, inflammation, angiogenesis, and many other biological processes that protect and repair the heart after myocardial injury. We have shown that the potency of allogeneic MSCs for myocardial repair and angiogenesis in nonhuman primates (NHPs) can be significantly improved by culturing the cells under hypoxic conditions before administration. Our results also suggested that the improvement is likely mediated by increases in the paracrine activity of the hypoxia-preconditioned cells, but the molecular factors that contributed to this increase were not identified.
METHODS In a murine MI model, cardiac function, infarct size, vascular density were measured by echocardiography, Masson staining and immunofluorescence. Aortic ring and tube formation were performed to evaluate angiogenic activity. Microarray and RNA sequencing were applied to determine the difference between exosomes from hypoxia-preconditioned MSCs (hpEXOs) and normoxia-cultured MSCs (nEXOs) and with these treated heart tissue.
RESULTS Cardiac function, infarct size, and vascular density were significantly greater in mice treated with hypoxia-preconditioned MSCs (hpMSCs) than with normoxia-cultured MSCs (nMSCs) and with hpEXOs than with nEXOs. miR-486-5p levels were significantly greater in hpEXOs than in nEXOs, which also confirmed to play a central role in exosomal mediated cardiac repair. Matrix metalloproteinase 19 (MMP19) levels were significantly lower in tissues from hpEXO-treated than nEXO-treated hearts and significantly greater in cardiac fibroblasts (CFs). miR-486-5p interacted with the 3' untranslated region of MMP19, and downregulated MMP19 expression in CFs, while both miR-486-5p upregulation and MMP19 silencing (MMP19 siRNA) in CFs increased the angiogenic activity of ECs cultured in the CF-conditioned medium. MMP19 silencing in CFs also reduced the cleavage of extracellular vascular endothelial growth factor (VEGF), and MMP19 cleaved VEGF in solution, while miR-486-5p upregulation significantly increased the potency of nEXOs for myocardial recovery and angiogenesis in an NHP MI model.
CONCLUSIONS In summary, the results presented in this report show that exosomal miR-486-5p is one of the primary pro-angiogenic paracrine factors produced by MSCs, and that it functions by downregulating MMP19 expression in fibroblasts and the cleavage of extracellular VEGFA. Furthermore, the exosomes produced by miR-486-5p – overexpressing MSCs significantly improved measures of cardiac function, infarct size, and angiogenesis when delivered to the infarcted hearts of NHPs without increasing the occurrence of arrhythmic complications. Collectively, these observations support additional investigations of the role of miR-486-5p, as well as the exosomes produced by other stem-cell populations, in myocardial regeneration.
GW31-e1214
Zhiru Zeng, Xianbao Liu, Jianan Wang
Second Affiliated Hospital, College of Medicine, Zhejiang University
OBJECTIVES Declined function of aged stem cells diminishes the benefits of autologous cell therapy for myocardial infarction. Mitochondrial dysfunction is associated with stem cell aging and it has received increasing attention as a target to restore aged stem cell function. Silent information regulator 1 (SIRT1) has a regulatory effect on mitochondrial dynamics. We have previously demonstrated SRT1720, a specific SIRT1 activator, protected aged human mesenchymal stem cells (hMSCs) against the extrinsic apoptotic pathway by upregulating FAIM1 and those cells pretreated with SRT1720 exhibited improved survival rate and achieved increased cardiac function in a rat model of myocardial infarction (MI). However, the role of mitochondria in SRT1720 mediated inhibition of apoptosis was not elucidated. The aim of the current study was to investigate the role of mitochondria in the anti-apoptotic effects of SRT1720 pretreatment of aged mesenchymal stem cells.
METHODS To study the effect of SRT1720 pretreatment on mitochondrial function, mitochondrial contents were evaluated by mitotracker staining, mtDNA measurements and expression of mitochondrial components and mitochondrial membrane potential was assessed by TMRM staining. Transmission electron microscopy was applied to assess mitochondrial morphology. As for in vivo studies, immunosuppressed Cynomolgus monkeys were subjected to myocardial infarction and treated with vehicle treated or SRT1720 pretreated aged hMSCs cells or DMEM. Cardiac function, cardiac cell apoptosis, angiogenesis and aged hMSCs engraftment were evaluated.
RESULTS Here we report that SRT1720 protects aged hMSCs against mitochondria apoptosis pathway by increasing mitochondrial biogenesis and function. We showed that SRT1720 pretreated aged hMSCs has reduced release of cytochrome C and caspase9 activation when subjected to H2O2 treatment and mitochondrial morphology was better preserved indicated by transmission electron microscopy. Mitochondria contents were compared between young and aged hMSCs by quantifying mtDNA level. Consistent with previous reports, aged hMSCs had decreased mitochondrial numbers and reduced expression of PGC1A and TFAM. We found that SRT1720 increased mitochondrial biogenesis of aged hMSCs, reflected by enhanced mtDNA levels, mitotracker staining, and expression of mitochondrial components. This resulted in higher mitochondrial respiratory capacity and oxidative phosphorylation (OXPHOS) efficiency. At 3 days after transplanted in the infarcted non-human primate hearts, SRT1720 pretreated aged hMSCs had more retention rate than vehicle treated controls. And histological analysis showed that cardiac cell apoptosis was attenuated in the SRT1720 group at 3 days post MI whereas no effect of the vascular density was observed at 3 months. Concomitantly, the recovery of cardiac contractile function at 3 months post MI in the SRT1720 group tended to be better than that in the vehicle group.
CONCLUSIONS SRT1720 promotes survival abilities of aged hMSCs in infarcted non-human primate hearts by increasing mitochondrial biogenesis and function. Although further research on the efficient strategies to rejuvenate aged hMSCs is required, the present study demonstrates that regulation of mitochondrial function through enhancing mitochondrial biogenesis is a potentially effective, low-cost, and stable treatment method to improve aged mesenchymal stem cell survival.
GW31-e1215
Jiabing Rong, Yinchuan Xu, Jianan Wang
Second Affiliated Hospital, Zhejiang University School of Medicine
OBJECTIVES Sepsis-induced myocardial dysfunction (SIMD) leads to severe mortality. The renin-angiotensin system (RAS) plays an important role in SIMD. Angiotensinogen (AGT) is the only known substrate of the RAS, and here we investigated the effects and mechanisms of AGT on the development of SIMD.
METHODS Male C57BL/6 mice were intraperitoneally injected with lipopolysaccharide (LPS) to induce sepsis and taken down 6 hours later. Cardiac function was evaluated by Vevo 2100 imaging system. Plasma levels of AGT, angiotensin II (AngII) and proinflammatory cytokines were detected by ELISA. Combination of AGT and its receptor, LDP receptor related protein1 (LRP1), was confirmed by ligand blotting in cardiac fibroblasts. Furthermore, RNA sequencing of cardiac tissue was performed to uncover the role of hepatic AGT on hearts after LPS challenge.
RESULTS Intraperitoneal injection of LPS significantly activated AGT expression both in liver and heart and elevated plasma AGT level. We further generated hepatocyte-specific AGT-deficient (hepAGT-/-) mice and cardiomyocyte-specific AGT-deficient (carAGT-/-) mice. Interestingly, we found that hepAGT-/- mice triggered resistance to SIMD. Contrarily, carAGT-/- mice had a similar phenotype with wild-type (WT) littermates. Depressed plasma AGT and AngII concentrations hinted hepatic deficiency of AGT may functioned via a circulatory AngII-dependent manner. However, WT mice infused with an AT1R antagonist, losartan, partially protected from cardiac dysfunction compared to hepatic AGT deficiency, which implied that liver-secreted AGT may have an another effect on heart via an AngII-independent pathway. Furthermore, we found cardiac AngII content of hepAGT-/- mice was comparable with WT littermates while IL-1β mRNA level in heart was lower, which proved there exited a pathway which IL-1β was involved in while AngII was not. Then, we discovered that hepAGT-/- heart showed a remarkably decreased AGT protein level. However, in carAGT-/- mice, cardiac AGT expression was restrained in mRNA level but had no change in protein level compared with their WT littermates. These facts implied that cardiac AGT was mainly derived from circulation rather than synthesized locally. Furthermore, we identified liver-derived AGT entered cardiac fibroblasts via LRP1-mediated endocytosis, which in turn activated NLRP3 inflammasome and improved IL-1β production.
CONCLUSIONS Hepatocyte-specific deficiency of AGT ameliorates SIMD via preventing cardiac fibroblastic LRP1-dependent endocytosis and then decreasing NLRP3 inflammasome expression, which alleviates IL-1β releasing. These findings provide potential therapeutic targets in liver to treat SIMD.
GW31-e1216
Feng Zhang1, Xuyang Fu1, Masaharu Kataoka2, Tian Liang1, Feng Gao1, Ning Liu1, Xiaoxuan Dong1, Xiaoyun Hu2, Wei Zhu1, Hong Yu1, Xingyang Hu1, Jianan Wang1, DaZhi Wang2, Jinghai Chen1
1Second Affiliated Hospital Zhejiang University School of Medicine
2Boston Children’s Hospital, Harvard Medical School
OBJECTIVES Cardiac fibrosis occurs in most cardiac disease, reduces cardiac muscle compliance, impairs both systolic and diastolic heart function and ultimately leads to heart failure. Long noncoding RNAs (lncRNAs) are recently emerging as important regulators of a variety of biological processes. However, little is known about the expression and function of lncRNAs in cardiac fibrosis. We aimed to identify cardiac fibroblasts (CFs)-enriched lncRNAs and investigate their role in regulation of cardiac fibrosis and heart function.
METHODS We performed microarray-based transcriptome profiling on mouse infarcted heart and from Microarray analysis we found a cardiac fibroblasts-enriched lncRNA, namely Cfast. We used lentivirus system to knockdown Cfast expression to investigate the functional regulation of Cfast in cardiac fibrosis in vivo and vitro. We further performed RNA pull down to find its downstream target protein for elucidating the molecular regulation of Cfast in cardiac fibrosis.
RESULTS Knockdown of Cfast in cardiac fibroblast, resulted in a significant down regulation in both mRNA and protein expression of fibrotic ECM-related gene. At the same time, we observed a significant decrease of α-SMA positive cells and a noticeable attenuation of the migratory ability of Cfast-silenced fibroblasts. After 28 days of MI, we found that mice with Cfast depletion exhibited significantly improved cardiac function. Immunohistochemical analysis showed that scars of Cfast knockdown mice contained more cardiomyocytes compared with control hearts. Histological analysis revealed that Cfast inhibition significantly reduced scar formation in infracted heart as well. We found that Cfast depletion significantly prevented isopropanol-induced cardiac pathological fibrosis in the heart. This protective evidence was also supported by the impact on expression of the molecular marker genes, as a significant reduction of Nppa, Nppb, and Myh7 and of cardiac fibrosis associated genes, Col1a1, Col3a1, Eln, and α-Sma upon Cfast depletion. Finally, we performed RNA pull-down assay to elucidate the molecular mechanism. Among analyzed lncRNAs binding partners, five proteins were detected as specifically interacted with Cfast. Nevertheless, considering the subcellular localizations and documented reports, we speculated COTL1 can act as a functional binding protein because COTL1 involves in TGFβ signaling pathway. To confirm the mass spectrometry data, we performed independent batch of Cfast RNA pull-down assay, and then used western blotting experiment to detect COTL1 after the lncRNA pull-down. The binding of COTL1 was clearly observed in Cfast pull-down but not in the control samples. The association of COTL1 with Cfast was further confirmed by RNA immunoprecipitation. Co-immunoprecipitation assays indicated that COTL1 competitively binds with TRAP1, leads to abrogate the interaction of TRAP1 with SMAD4 and thus suppresses the formation of SMAD2/3/4 complex.
CONCLUSIONS We identified a novel cardiac fibroblast-enriched lncRNA, named Cfast, as an important regulator in the process of cardiac fibrosis. Cfast depletion clearly exhibits protective effects from pathological fibrotic remodeling and improves cardiac function upon pathological stress. Cfast exerts its function via binding COTL1 and consequently affects TRAP1/SMAD mediated down cascades of fibrotic signal. Our study indicates that Cfast may represent a potential target in antifibrotic RNA therapy in heart diseases.
GW31-e1217
Yue Tao, Rongrong Wu, Xinyang Hu, Jianan Wang
Second Affiliated Hospital of Zhejiang University School of Medicine
OBJECTIVES Bone marrow mesenchymal stem cells (MSCs) transplantation is a new hope for the treatment of myocardial infarction, but low survival rate of engrafted MSCs mainly limits the therapeutic effect of MSCs. Illustrating mechanism of stem cells survival and improving the therapeutic effect of engrafted cells are primary issues remained to be resolved. In this study, long non-coding RNA was used to improve the anti-apoptotic ability of MSCs and the mechanism involved was explored.
METHODS After 24 hours under normal culture of hypoxia treatment, MSCs were under the screening process of lncRNA sequencing, filtered for lncRNA LCUAT1. By lentivirus editing LUCAT1 level of MSCs and injecting the MSCs into the border zone of the infarcted heart, we aimed to find out whether LUCAT1 influences the apoptosis of MSCs and the therapeutic effect of MSCs for myocardial infarction. GFP staining 3 days after myocardial infarction, echo detection and Masson staining 28 days after myocardial infarction were conducted. Furthermore, through mass spectrometry analysis of proteins that were co-IP by LUCAT1 and RIP experiment, we screened regulatory proteins in the nucleus which LUCAT1 could bind to. Synthesis of the results from mass spectrometry and transcriptome sequencing of MSCs after LUCAT1 knocked down, we singled out the specific target gene regulated by LUCAT1. With CHIP assay and luciferase assay, we explored LUCAT1 may manipulate the apoptosis of MSCs by recruiting regulatory protein to target gene promoter.
RESULTS We found that LUCAT1 knock down decreased cell apoptosis resistance while LUCAT1 over expression showed opposite results. GFP staining showed that survival rate of the MSCs three days post injection was reduced, and the echo results 28 days after the myocardial infarction indicated that the cardiac function was worse than control group. In addition, the Masson staining results after 28 days showed that the scar area of heart was increased. Overexpression of LUCAT1 can enhance the anti-apoptotic ability of cells and the therapeutic effect after myocardial infarction. It was found that LUCAT1 could bind to a lot of regulatory proteins in the nucleus, RIP results showed LUCAT1 had strong binding ability with JMJD6. Through transcriptome sequencing of MSCs after LUCAT1 knock down, the expression of FOXQ1, an anti-apoptotic protein, was significantly decreased compared with control group. When LUCAT1 was overexpressed, FOXQ1 expression increased significantly. Lentivirus knockdown of JMJD6 in mesenchymal decreased FOXQ1 expression, but overexpression had no effect on FOXQ1 expression. Luciferase experiment verified that JMJD6 could bind to the promoter region of FOXQ1. CHIP detection detected that JMJD6 act in FOXQ1 promoter region on the methylation sites H3R2me2a and H4R3me2s reported. It was also found that the methylation level of these two sites increased when LUCAT1 was knocked down, while the methylation enrichment of H4R3me2s decreased when LUCAT1 was overexpressed and H3R2me2a showed no significant change.
CONCLUSIONS LUCAT1 can improve the viability of MSCs and enhance its therapeutic effect on myocardial infarction by recruiting JMJD6 to the anti-apoptotic protein FOXQ1 promoter, affecting the methylation enrichment level of H4R3me2s in the promoter region and thus changing FOXQ1 expression level.
GW31-e1220
Qingnian Liu, Yue Tao, Hao Ding, Changchen Xiao, Yu Zhou, Cheng Ni, Changle Ke, Jingyi Wang, Rongrong Wu, Lin Fan, Xianpeng Wu, Jing Zhao, Yan Wu, Xinyang Hu, Jianan Wang
Second Affiliated Hospital, Zhejiang University School of Medicine
OBJECTIVES Myocardial infarction (MI) is characterized by cardiac dysfunction and increased cardiomyocyte death, induced mainly by apoptosis. Using an unbiased transcriptome analysis, we identified flavin containing monooxygenase 2 (FMO2) as one of the top-ranked genes involved in the process of MI. In this study, we investigate the roles of FMO2 in ischemic injury and its potential mechanisms. FMO2 exhibits the cardiac protection from MI injury.
METHODS Male SD rats receiving either adeno-associated virus serotype 9 containing FMO2 shRNA particles (AAV-shFMO2) or FMO2 (AAV-FMO2), and FMO2 knockout rats were subjected to myocardial infarction surgery. Cardiac function, fibrosis, and apoptosis were examined in these rats and related cellular and molecular mechanisms were investigated.
RESULTS Cardiac ischemia injury was associated with significant increases of FMO2 levels both in ex vivo and in vivo models. Loss of FMO2 significantly enhanced cardiomyocyte apoptosis and deteriorated cardiac function accompanied by augmented infarct size in infarcted rat hearts, while elevated expression of FMO2 exhibited the opposite results. Mechanically, located on the ER membrane, FMO2 inhibited activation of ER stress-initiated apoptotic proteins including caspase 12 and C/EBP homologous protein (CHOP), via down-regulating upstream unfolded protein response (UPR) pathway. Furthermore, we found that FMO2, as a novel chaperone in ER, directly catalyzed disulfide-bond synthesis to facilitate proteins folding. Finally, structure analysis of FMO2 revealed the active site GVSG for disulfide-bond catalysis, which was confirmed by the molecular docking experiment of GSH with FMO2. However, FMO2 with GVSG mutation failed to catalyze disulfide-bond formation and lost protection from ER stress or apoptosis in cardiomyocytes.
CONCLUSIONS FMO2 confers cardiac protection from ischemic damage due to improved cardiomyocyte apoptosis through UPR pathway, which is mediated by disulfide-bond catalysis at GVSG active site. Our findings uncover a novel FMO2-involved regulatory mechanism which could serves as a potential therapeutic target for ischemic cardiovascular diseases.
GW31-e1221
Wangxing Hu, Rongrong Wu, Chenyang Gao, Xianbao Liu, Jianan Wang
Second Affiliated Hospital, Zhejiang University School of Medicine
OBJECTIVES Calcific aortic valve disease (CAVD) is the most common valvulopathy in developed countries and is characterized by inflammation, extracellular matrix (ECM) remodeling and calcification, leading a narrowing of the valve and the consequential obstruction of the cardiac outflow. A lot work has shown that valve interstitial cells within the aortic valve cusps when stimulated could differentiate toward an osteoblast-like cell and deposit bone-like matrix that leads to leaflet stiffening and calcific aortic valve stenosis. However, the mechanisms that promote pathological phenotypes in valve interstitial cells are still not clear. Nuclear receptors regulate the transcription of genes involved in mitochondrial biogenesis in a tissue-specific manner. Accumulating evidence has suggested that NRs make contributions to calcific vascular and valvular disease. Estrogen-related receptor α (ERRα) is an orphan nuclear hormone receptor capable of regulating transcription of genes involved in multiple cellular and physiological processes. However, the role of ERRα in valve calcification has not been investigated to date. The aim of the present study was to examine the role of ERRα in aortic valve calcification.
METHODS Aortic valve leaflets were collected from CAVD patients (N=10) undergoing aortic valve replacement, and Control samples (no calcified aortic valve tissue) were taken from age-matched patients who had severe insufficiency without calcification (n=7) and patients who underwent heart transplantation due to late stage of cardiomyopathy (n=3). We compared the protein level of ERRα by Western blot and immunohistochemistry between the non-calcific valve and the calcific ones. In vitro study, valve interstitial cells (VICs) were isolated from the control aortic valve cusps and cultured in calcifying medium to induce calcification. Treated with 10 μM XCT790 or ERRα small interfering RNA silencing to inhibit ERRα or overexpression of ERRα by lentivirus, the samples were assessed for calcium nodule formation, and calcific markers using alizarin red staining, ALP enzyme activity assay, Western blot and qPCR. The effect of ERRα on osteoblastic transdifferentiation and its downstream pathway were also studied.
RESULTS In this study, we have found that the protein level of ERRα is upregulated in CAVD samples versus the controls; moreover, during osteogenic differentiation of hVICs, the expression of ERRα is increased as the cultural time increased. Furthermore, we found that inhibition of ERRα prevented OM-induced upregulation of Runx2 and ALP. The ALP activity was markedly attenuated by ERRα silencing. Alizarin red staining revealed that shRNA-mediated ERRα knockdown markedly suppressed calcified nodules formation. Meanwhile, the content of calcium was significantly reduced. We also proved that ERRα overexpression enhances osteogenic potential of hVICs. In addition, RNA sequencing results suggested that heme oxygenase-1 (Hmox1) was a downstream target of ERRα and it was further confirmed by western blot. Additionally, we also found that downregulation of Hmox1 with shHmox1 efficiently reversed the inhibition of calcification induced by ERRα shRNA in hVICs. ChIP-qPCR and luciferase assay indicated that Hmox1 was negatively regulated by ERRα.
CONCLUSIONS The present results indicate that knockdown of ERRα could impair the osteoblastic transdifferentiation of VICs, suggesting that inhibition of ERRα is a potential therapeutic strategy for the prevention of aortic valve calcification.
GW31-e1244
Chong Huang1, Xiong Guo1,2, Ling Tao1
1Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi’an, Shaanxi, China
2Department of Cardiology, No. 952 Hospital of People’s Liberation Army, Golmud, Qinghai, China
OBJECTIVES The study is designed to explore the role BCAA catabolic defect plays in the coronary microvascular dysfunction (CMD) and diabetic cardiomyopathy (DCM) in T2DM.
METHODS Genetically BCAA catabolism-defective mice (PP2Cm-/-) and BCAA catabolism-enhanced mice (PP2CmOE/OE) were built and T2DM was induced in 3 groups of mice (WT, PP2Cm-/-, PP2CmOE/OE) by high-fat diet feeding. In the diabetic duration, the cardiac function was monitored in each group of mice. After 12 weeks, the microvascular density (MVD) in heart tissue and leaky phenotype of the myocardial microvessel in each group of mice were detected with immunohistochemical staining. Also, cell survival, growth, migration and vascular permeability were tested in mouse coronary ECs (MCECs) isolated from each group of mice in vitro.
RESULTS PP2Cm-/- mice manifested deteriorated loss of cardiac function compared with WT mice. The MVD in heart tissue was significantly lower and myocardial microvessel leakage was markedly increased in PP2Cm-/-. Meanwhile, In vitro, the MCECs isolated from PP2Cm-/- showed aggravated cell function loss of proliferation, migration and increased vascular permeability compared with WT. However, PP2CmOE/OE mice manifested preserved cardiac function and MCECs function during T2DM. As to the mechanism, BCAA treatment markedly reduced expression of STIM1 and thereby inhibited mTORC2-Akt1 passway in MCECs. Restoration of STIM1 expression and mTORC2-Akt1 regain cell function of MCECs.
CONCLUSIONS BCAA catabolic defect promotes CMD and DCM in T2DM. Downregulation of STIM1 in MCECs induced by catabolic defect of BCAA and resulting inhibition of mTORC2-Akt1 were the potential mechanism.
GW31-e1248
Qi Li, Bochuan Li, Yi Zhu
Tianjin Key Laboratory of Metabolic Diseases; Key Laboratory of Immune Microenvironment and Disease (Ministry of Education); Collaborative Innovation Center of Tianjin for Medical Epigenetics and Department of Physiology and Pathophysiology, Tianjin Medical University, Tianjin, China
OBJECTIVES CEMIP (cell-migration-inducing and hyaluronan-binding protein) is a newly discovered macromolecular protein. It plays an important role in the proliferation and migration of tumors. Bioinformatics analysis suggests that it is highly expressed in smooth muscle cells. Vascular smooth muscle cells are critical in maintaining vascular homeostasis. We aim to investigate the role of CEMIP in proliferation and contraction of vascular smooth muscle cells (VSMCs).
METHODS In vivo, we established tamoxifen-inducible smooth muscle specific CEMIP-deficient (CEMIPΔsmc) mice. Both CEMIPfl/fl mice and CEMIPΔsmc mice were subjected to wire-guided common carotid injury to evaluate the neointima formation. Simultaneously, blood pressure was monitored by non-invasive tail-cuff system. Aortas and second-order mesenteric arteries were isolated from male mice and were used to evaluate the vascular tone by isometric force measurement in wire myograph. In vitro, flow cytometry, ki67 staining, cell count, cell scratch and transwell assay were used to detect elicited human aortic smooth muscle cells (HAoSMCs) proliferation and migration. Contractile gel assay was used to detect contraction. Small interfering RNA (siRNA) transfection, real-time PCR and western blot were utilized for further verifying the detailed effects of CEMIP on VSMCs phenotypes.
RESULTS CEMIP deficiency could significantly inhibit the platelet-derived growth factor-BB (PDGF-BB) induced VSMCs proliferation and migration. Compared with CEMIPfl/fl mice, the neointimal hyperplasia of CEMIPΔsmc mice was dramatically reduced. Meanwhile, loss of CEMIP also reduced VSMCs contractile phenotype-dependent markers expression accompanied with less related contractile area. This phenomenon was also confirmed in CEMIPΔsmc mice aortas. In accordance, the blood pressure of CEMIPΔsmc mice was lower than littermate control mice under physiological state and angiotensin II-induced hypertension. The vascular ring experiments also confirmed that the CEMIP reduction could markedly inhibit phenylephrine-induced vasoconstriction, while not response to ET-1, PGF2α and 5-hydroxytryptamine.
CONCLUSIONS Our results demonstrate the significance of CEMIP in regulating vascular smooth muscle proliferation and vessel tension maintenance.
GW31-e1249
Hao Wang, Peng Zhang, Xiahuan Chen, Meilin Liu
Department of Geriatrics, Peking University First Hospital, Beijing 100034, People’s Republic of China
OBJECTIVES Activin A, a member of the transforming growth factor-β superfamily, has been reported to play important roles in regulating macrophages foam cell formation. However, the underlying mechanisms remain to be elucidated. The purpose of this study is to investigate the effects of activin A on expressions of cholesterol influx/efflux transporters and underlying molecular mechanisms.
METHODS The effects of activin A on Dil-ox-LDL uptake by RAW 264.7 macrophages were analyzed by confocal microscopy and flow cytometry. The mRNA and protein levels of cholesterol influx/efflux transporters were examined by RT-qPCR and western blot analysis, respectively. To investigate whether ALKs are involved in the activin A-mediated inhibition of foam cell formation, macrophages were pretreated with selective ALKs inhibitor, SB431542, then co-treated with ox-LDL and activin A. The involvement of SMAD2, SMAD3 and SMAD4 were confirmed by transfection with specific siRNAs.
RESULTS The results showed that activin A decreased Dil-ox-LDL uptake in RAW 264.7 macrophages. The mRNA and protein levels of SR-A1 was significantly down-regulated in activin A treated macrophages, while the expression of ABCA1 and ABCG1 was up-regulated. Pre-treatment with SB431542 reversed the activin A-mediated inhibition of foam cell formation. Knockdown of SMAD2 reversed the activin A-mediated reduction of Dil-ox-LDL uptake and SR-A1 expression. However, knockdown of SMAD3 or SMAD4 did not have such effect. Meanwhile, knockdown of either SMAD2, SMAD3 or SMAD4 reversed the activin A-mediated upregulation of ABCA1 and ABCG1.
CONCLUSIONS Activin A inhibits ox-LDL-induced foam cell formation in RAW 264.7 macrophages. The molecular mechanism may be related to the down-regulation of SR-A1 and up-regulation of ABCA1 and ABCG1 through ALK-SMAD signaling pathway.
GW31-e1263
Ke Lin, Weijian Huang
The Key Laboratory of Cardiovascular Disease of Wenzhou, Department of Cardiology, The First Affiliated Hospital of WenZhou Medical University, WenZhou
OBJECTIVES LCZ696 was confirmed to be an effective mixture for numerous heart failure, whether it would be effective for alcoholic cardiomyopathy was still unknown. In this study, we aimed to elucidate the role of LCZ696 in ethanol induced cardiac injury and to explore possible mechanisms.
METHODS In vitro model of ethanol induced cardiac injury was established as previously reported. In our study, 10 μM and 20 μM LCZ696 was pretreated for 1 hour, and then 200 mM ethanol was added into the medium for 8 h. Cells were sacrificed for further study. Total protein was collected, and the expression of Bax, Bcl-2, caspase-3, p-p65, p-65 and GAPDH was detected using western blotting. And the expression of TNF-α, IL-1β and IL-6 was detected using qPCR.
RESULTS After the treatment of 200 mM ethanol for 8 h, expression of caspase-3 and Bax was elevated while the expression of Bcl-2 was decreased, indicating the cardiomyocytes apoptosis. However, treatment of LCZ696 could partially reverse the condition with declined expression of caspase-3, Bax and increased Bcl-2. Besides, NF-κB signaling was also detected. Two hundred millimolar ethanol for 6 h could significantly activate NF-κB with the increasing phosphorylation of p65. And the pretreatment of LCZ696 could effectively inhibit the activation, with the decrease of p-p65. Similarly, pretreatment of LCZ696 could also inhibit ethanol induced overexpression of TNF-α, IL-6 and IL-1β.
CONCLUSIONS In this study, we confirmed that LCZ696 could protect cardiomyocytes from ethanol induced cellular apoptosis and inflammation.
GW31-e1264
Jie Yang, Lan Huang
Department of Cardiology, the Second Affiliated Hospital, Third Military Medical University (Army Medical University), Chongqing, China
OBJECTIVES Statins play a major role in reducing circulating cholesterol levels and are widely used to prevent coronary artery disease and stroke. Although they were recently confirmed to up-regulate mitophagy, little is known about the molecular mechanisms underlying statin-induced mitophagy. Whether mitophagy induction contributes to endothelial progenitor cell (EPC) proliferation also remains to be elucidated. Here, we explored the role and mechanism underlying statin (pitavastatin, PTV)-activated mitophagy in EPC proliferation using atherosclerotic mice.
METHODS ApoE-/- mice were fed a high-fat diet for 8–16 weeks to induce atherosclerosis. GFP-mRFP-LC3 and mt-Keima were used to evaluate autophagy and mitophagy flux, respectively. 3-MA and Atg7 silencing were used to inhibit autophagy. MMP assay kit, JC-1 and MitoSox red were employed to measure mitochondrial function. Calcium concentration was analyzed in cytoplasm and mitochondria in intact cells through Fluo-3-AM and Rhod2-AM, respectively. Cell proliferation was checked by the xCelligence Real-Time Cell Analyzer instrument. Mitochondrial morphology and autophagosomes were observed by transmission electron microscopy. Western blots were employed to detect the expression of protein in cytoplasm or mitochondria.
RESULTS EPC proliferation decreased in ApoE-/- mice, and was accompanied by mitochondrial dysfunction and mitophagy impairment via the PINK1-PARK2 pathway in comparison to those of normal diet mice. PTV reversed mitophagy and reduction in proliferation. Mitophagy inhibition by 3-MA treatment or by silencing Atg7 blocked PTV-induced proliferation improvement, suggesting that mitophagy contributed to the EPC proliferation increase. PTV elicited mitochondrial calcium release into the cytoplasm and further phosphorylated CAMK1. CAMK1 served as a PINK1 kinase to reverse the PINK1-PARK2 pathway of mitophagy and mitochondrial dysfunction in EPCs derived from atherosclerotic mice.
CONCLUSIONS Our findings describe a novel molecular mechanism of mitophagy activation, where mitochondrial calcium release promotes CAMK1 phosphorylation of threonine177 before phosphorylation of PINK1 at serine228, which recruits PARK2 and phosphorylates its serine65 to activate mitophagy. Our results further account for the pleiotropic effects of statins on the cardiovascular system and provide a promising and potential therapeutic target for atherosclerosis.
GW31-e1267
Yujie Yang, Jinlong He
Tianjin Medical University
OBJECTIVES Disturbed flow induces endothelial dysfunction and contributes to uneven distribution of atherosclerotic plaque. Emerging evidence suggests that harmine, a natural small molecule in extracts of Peganum harmala, has potent beneficial activities. Here, we investigated whether harmine has an atheroprotective role under disturbed flow and the underlying mechanism.
METHODS Mice of ApoE-/-, LDLR-/- and endothelial cell (EC)-specific overexpression of Yes-associated protein (YAP) in ApoE-/- background were fed with a western diet and given harmine for 4 weeks. Atherosclerotic lesion size, cellular composition and expression of inflammatory gene in the aortic roots were observed. Human umbilical vein ECs (HUVECs) were treated with oscillatory shear stress (OSS) and harmine, and also used for proteomic analysis.
RESULTS Harmine retarded atherogenesis in both ApoE-/- and LDLR-/- mice by inhibiting the endothelial inflammatory response. Mechanistically, harmine blocked OSS-induced YAP nuclear translocation and EC activation by reducing phosphorylation of YAP at Y357. Overexpression of endothelial YAP blunted the beneficial effects of harmine in mice. Proteomic study revealed that tyrosine-protein phosphatase non-receptor type 14 (PTPN14) could bind to YAP. Moreover, harmine increased PTPN14 expression by stabilizing its protein level and inhibiting its degradation in proteasome. PTPN14 knockdown blocked the effects of harmine on YAPY357 and EC activation. Finally, overexpression of PTPN14 mimicked the effects of harmine and ameliorated atherosclerosis in a partial ligation mouse model.
CONCLUSIONS Harmine alleviated OSS-induced EC activation via a PTPN14/YAPY357 pathway and had a potent atheroprotective role.
GW31-e1274
Ruiyu Wang, Jing Huang
Department of Cardiology, the Second Affiliated Hospital of Chongqing Medical University
OBJECTIVES Excessive oxidative stress is a major causative factor of endothelial dysfunction in hypertension. As an endogenous pro-oxidant, thioredoxin-interacting protein (TXNIP) contributes to oxidative damage in various tissues. The present study was aimed to investigate the role of thioredoxin-interacting protein (TXNIP) in mediating hypertension-induced endothelial dysfunction.
METHODS In vivo, 30 rats were divided into the control, sham, and hypertension groups (n=10 per group) randomly, and the experimental model of hypertension was conducted by two kidney one clip (2K1C) surgery. Blood pressure and heart rate were monitored regularly. At 4 weeks after the 2K1C surgery, multiple vascular tissues (carotid artery and aorta) and plasma were harvested for TXNIP and plasma angiotensin (Ang) II concentration detection, respectively. Subsequently, 32 rats were divided into the sham, sham+resveratrol (as a TXNIP inhibitor), hypertension and hypertension+resveratrol groups (n=8 per group). Endothelium-dependent vasodilation, DHE staining and Western blot were performed in the aorta of rats. In vitro, primary human aortic endothelial cells (HAECs) were stimulated with human Ang II to mimic endothelial injury in hypertension. Losartan (an Ang II type I receptor blocker) and PX-12 (an inhibitor of thioredoxin) were administered to HAEC in the presence of Ang II stimulation. In addition, TXNIP-specific siRNA and thioredoxin (TRX)-loaded GV230 plasmid were constructed and transfected into HAEC. The oxidative stress, cell apoptosis, eNOS activation and intracellular nitric oxide (NO) production were evaluated.
RESULTS In vivo, TXNIP was upregulated in vascular endothelial cell in parallel with elevated plasma Ang II in hypertensive rats. Inhibition of TXNIP by resveratrol improved the endothelium-dependent vasodilation, restored eNOS expression/activation and suppressed systemic and vascular oxidative stress in hypertension rats. In vitro, TXNIP was substantially increased in Ang II-stimulated HAECs in a time and concentration-dependent manner, and such increase could be partly reversed by Losartan. TXNIP knockdown by siRNA alleviated Ang II-induced oxidative stress and cell apoptosis as evidenced by a reduction of ROS generation and alterations in the protein levels of NOX2, NOX4, SOD2, ASK1, cleaved-Caspase 3 and the ratio of BCL2/BAX. In addition, inhibition of TXNIP effectively rescued the impaired NO production and maintained the ratios of p-eNOS (ser 1177)/eNOS and p-AKT (ser 473)/AKT in Ang II-treated HAECs. Importantly, TXNIP knockdown significantly upregulated TRX expression and promoted the nuclear translocation of TRX to activate the DNA repair related-transcription factors (AP-1 and REF-1). However, inhibiting TRX with PX-12 completely blunted the protective effect of silencing TXNIP on HAECs, and overexpression of TRX effectively upregulated p-eNOS (ser 1177)/eNOS ratio and improved NO production in HAECs under Ang II stimulation.
CONCLUSIONS Our study, for the first time, suggests that TXNIP contributes to oxidative stress and endothelial dysfunction in hypertension, and these effects are dependent on the antioxidant capacity of TRX. Maintaining vascular TXNIP/TRX homeostasis may be beneficial in the exploration of new treatments for hypertension and other cardiovascular diseases.
GW31-e1281
Yupeng Wang, Yanyan Liu, Lingyun Zu
Peking University Third Hospital
OBJECTIVES We compared the success rate on the first attempt, the procedure time, operator satisfactions with the instruments used, the pain score and complications between intraosseous (IO) access and central venous catheterization (CVC) in critically ill Chinese patients for whom two attempts to establish peripheral venous access had failed or for whom treatment was delayed.
METHODS Experimental minipigs were chosen for this study and were randomly divided into intraosseous access and central venous catheter groups (n=4 for each group). Cecal ligation and puncture was performed to establish the septic shock model. Immediately after the septic shock model was established, the intraosseous access group received norepinephrine via tibial intraosseous access infusion, whereas the central venous catheter group received norepinephrine via intravenous infusion of the internal jugular vein. The vital signs, blood pressure recovery time, post-septic shock survival time, and total survival time of the minipigs were recorded.
RESULTS There were no statistically significant differences between the two groups in terms of their vital signs (blood pressure, heart rate, temperature), blood routine, myocardial enzymes, kidney function, liver function, lactate, interleukin-6 and interleukin-8 at pre-Cecal ligation and puncture, 4 h post-Cecal ligation and puncture, 10 h post-Cecal ligation and puncture, and shock onset (P>0.05). There was no significant difference in the time needed to establish the septic shock model between the intraosseous access and central venous catheter groups [13.5 (10.5–25.5) vs. 14.5 (10.5–20.0) hours, P=0.886]. There was no significant difference in the mean arterial pressure at shock onset between the intraosseous access and central venous catheter groups [59.5 (57.0–62.0) vs. 61.0 (59.0–63.0) mmHg, P=0.343]. There was no significant difference in blood pressure recovery time between minipigs receiving intraosseous access and central venous catheter norepinephrine infusion [40 (20–60) vs. 30 (20–60) minutes, P=0.686]. The cumulative norepinephrine dose during the period of blood pressure recovery time did not differ significantly between the intraosseous access and central venous catheter groups [10.0 (3.0–21.0) vs. 6.5 (3.0–21.0) μg/kg, P=0.686]. There was no significant difference in the mean arterial pressure increase during the period of blood pressure recovery time between the intraosseous access and central venous catheter groups [14.5(10.0–24.0) vs. 13.0 (10.0–17.0) mmHg, P=0.686]. The two groups did not show statistically significant differences at 6 h after norepinephrine infusion in terms of vital signs, blood routine, myocardial enzymes, kidney function, liver function, lactate, interleukin-6, and interleukin-8 (P>0.05). There were no significant differences in the post-septic shock survival time [7.75 (6.50–10.00) vs. 8.75 (6.50–14.00) hours, P=0.686] and total survival time [21.25 (16.50–35.50) vs. 25.75 (19.50–26.50) hours, P=0.686] of minipigs in the intraosseous access and central venous catheter groups.
CONCLUSIONS In the experimental minipig model of septic shock, tibial intraosseous access and central venous catheter infusion of norepinephrine achieved same therapeutic effect on hypotension and survival time. If peripheral vascular access cannot be established during septic shock, intraosseous access norepinephrine infusion can be used to correct the hypotensive state.
GW31-e1286
Shenghuan Yu1,2,3, Long Zeng1,2,3, Wei Wei1,3
1Department of Cardiology, Guangdong Cardiovascular Institute
2Guangdong Cardiovascular Institute, Key Laboratory of Clinical Pharmacology, Guangdong Provincial People’s Hospital Medical Research Center, Guangdong Academy of Medical Sciences
3School of medical, South China University of Technology
OBJECTIVES To investigate the role of co-transcriptional activator p300 in atrial fibrosis caused by hydrostatic pressure.
METHODS Collect the left atrial appendage tissue of three groups of patients with sinus rhythm, simple atrial fibrillation, hypertension and atrial fibrillation, and detect the expression of p300 protein and fibrotic factors Collagen Type I Alpha 1 (Col1A1), Collagen Type III Alpha 1 Col3A1, matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP9), Smad3, p-Smad3 and transforming growth factor-β (TGF-β) in the three groups of left atrial appendage tissue by western blot. Primary cultured atrial appendage fibroblasts of 8–12 weeks C57BL/6 mice, and cultured with 0 mmHg, 20 mmHg and 40 mmHg hydrostatic pressure; further, the fibroblasts cultured under 0 mmHg and 40 mmHg pressure were treated with p300 HAT specific inhibitor curcumin and p300 interfering RNA, respectively, and the changes in the expression of p300 protein and fibrosis indicators in the cells were detected.
RESULTS The expressions of p300 protein and fibrosis indicators in patients with hypertension with atrial fibrillation were significantly higher than those of patients with atrial fibrillation or sinus rhythm (P<0.05). Under high hydrostatic pressure, the expressions of p300 protein and fibrosis indicators of mouse atrial fibroblasts increased and were gradient-dependent (P<0.05). After addition of p300 HAT-specific inhibitor curcumin and p300 interfering RNA, the increase of p300 and fibrosis indicators protein expression was reversed (P<0.05).
CONCLUSIONS p300 is involved in atrial fibrosis induced by high hydrostatic pressure.
GW31-e1289
Yue Yuan, Xuejie Han, Xinbo Zhao, Xin Bi, Yongtai Gong, Xiaoxu Duan, Yue Li
The First Affiliated Hospital of Harbin Medical University
OBJECTIVES Atrial glusose metabolism is dysregulated in atrial fibrillation (AF), but its precise molecular mechanism and its role in the progression of AF still remain unclear. Recent studies have reported that succinylation plays an important role to regulate cellular glucose metabolism. We hypothesize that downregulation of Sirt5 could induce PDH succinylation and suppress its activity, which promotes dysregulation of glucose metabolism resulting in atrial electrical and structural remodeling and the occurrence and persistence of AF.
METHODS We use the metabonomics, transgenic animal model, comprehensive techniques of bioinformational and functional biology to elucidate the role and mechanism of Sirt5 mediated succinylation in atrial abnormal glucose metabolism and progression of atrial fibrillation, by targeting regulation of PDH.
RESULTS We previously found that (1) The level of succinylated PDH, the essential enzyme to glucose metabolism, markedly increased in atrial tissues of AF patients accompanied with the downregulation of Sirt5, the key factor of succinylation. (2) Sirt5 knockout increased the level of PDH succinylation in atrial tissues, which resulted in the incidence and persistence of AF. (3) Overexpression of Sirt5 could prevent tachypacing-induced PDH succinylation, improve glucose metabolism and inhibit apoptosis and ion channel dysregulation in atrial myocytes.
CONCLUSIONS Sirt5 dependent succinylation of PDH drives the dysregulation of glucose metabolism resulting in atrial remodeling and the occurrence and persistence of AF.
GW31-e1291
Bing Dong, Wenhao Xia
The First Affiliated Hospital of Sun Yat-Sen University
OBJECTIVES Previous clinical studies have suggested that beyond the on-target of lowering blood glucose, SGLT2 inhibitors offers a significant reduction in major cardiovascular events in type 2 diabetes mellitus (T2DM) patients. Endothelial progenitor cells (EPCs) has proven great potential for endothelial injury repairing in vascular complications. This study aimed to evaluate the effect of dapagliflozin (DAPA) on EPCs in diabetes and the underlying mechanism involved with a focus on inflammation and oxidative stress.
METHODS This study enrolled 62 T2DM patients (DM group) and 60 matched healthy control participants (Control group). The vasculogenic capacity of EPCs were compared in vitro (migration, adhesion, tube formation abilities) and in vivo (limb ischemia model). Inflammation marker genes (IL-1β, IL-8, TNF-α and MCP-1) and oxidative stress genes (SOD-2, HO-1, TXN and CAT) expression were determined by RT-PCR. The AMPK signaling were tested by Western blot.
RESULTS T2DM EPCs demonstrated a declined in vitro and in vivo vasculogenic capacity of EPCs Inflammation and oxidative stress maker genes expression were significantly elevated in T2DM, which was accompanied with decreased phosphorylation of AMPK. DAPA restored AMP/ATP ratio to trigger AMPK activation and decreased the level of inflammation and oxidative stress, which ultimately leads to restored vasculogenic capacity of EPC. Moreover, the enhanced functions of EPCs by DAPA were blocked by AMPK inhibitors.
CONCLUSIONS Dapagliflozin rescues vasculogenic capacity of endothelial progenitor cells in Diabetic Limb Ischemia via AMPK-mediated inhibition of inflammation and oxidative stress. Up-regulation of AMPK signaling by DAPA may be a novel therapeutic intervention for vasculogenic capacity of EPCs. Our findings provide at least a partial explanation for the mechanism underlying the DAPA off target effect of vascular protective.
GW31-e1294
Hongye Chen, Zhenjie Liu, Yong Sun, XuanHao Wang, HaiQiong Zhen, LianLian Zhu, XiangMin Kong, Wei Zhu, Jianan Wang
The Second Affiliated Hospital, Zhejiang University School of Medicine
OBJECTIVES NADH dehydrogenase (ubiquinone) 1 alpha subcomplex subunit 13 is a subunit of Mitochondrial Respiratory Complexes I, namely NDUFA13. Our previous study shows that a moderate downregulation of NDUFA13 could protect against apoptosis and decrease cardiac ischemia reperfusion injury by generating a mild increase in ROS. This indicates that NDUFA13 might be a potential therapeutic target for treating cardiac injury. However, it remains unexplored regarding its role in vascular homeostasis. Therefore, our present study is designed to investigate the expression profile of NDUFA13 in an existing aortic aneurysm, and smooth muscle cell (SMC) specific NDUFA13 knock-out mice were established to study its function and mechanism in pathological process of vascular injury.
METHODS Immunohistochemistry staining of NDUFA13 was conducted in human and mouse aortic aneurysm tissues compared with normal aorta. Bioinformatic analyze was applied to find the expression profile of NDUFA13 in human vascular disease including aneurysm, dissection and hereditary vascular disease. NDUFA13fl/fl mice were crossed-bred with SM22-CreERT+ mice to generate SM22-CreERT+ NDUFA13fl/fl mice and SM22-CreERT- NDUFA13fl/fl mice. By injecting tamoxifen at 75 mg/kg/d for 5 consecutive days, NDUFA13 was successfully knocked out. Twelve to fourteen weeks old male mice underwent abdominal aneurysm induction by elastase (>4.0 U/mg) perfusion for 10 minutes. After 14 days, aorta were gathered for general imaging, diameter measurement, hematoxylin-eosin (H&E), Verhoeff Van Gieson (EVG) staining and elastin integrity grading score. Another model was also established and utilized to induce thoracic aortic aneurysm and dissection in 3–4 weeks old mice by water intake of 3-Aminopropionitrile fumarate salt (BAPN) at 1 g/kg/Day for 4 weeks. Aneurysm size, formation and death rate, histomorphology data were analyzed. In vitro study was conducted in MOVAS and SMC to explore the underline mechanism in metabolism and programmed cell death.
RESULTS NDUFA13 expression is dramatically increased in aortic aneurysm (P<0.001), both in human and mouse aneurysm tissue. Bioinformatic scan reveals downregulation of NDUFA13 in human Marfan’s syndrome. Aneurysm alleviation is significant in SM22CreERT+ NDUFA13fl/fl mice with a mean diameter of 0.796 mm compared with 1.157 mm in SM22CreERT- NDUFA13flox/flox mice (P<0.01). Better vascular integrity in H&E staining and less elastin fragmentation in EVG staining are observed after smooth muscle cells specific NDUFA13 knock out, and elastin integrity score is significantly decreased in knock-out mice (P<0.01). In BAPN induced aneurysm and dissection, aneurysm attenuation is observed at both aneurysm size (P<0.05), vascular integrity and aneurysm formation rate (P<0.05). After knockdown of NDUFA13 in MOVAS and SMC, the apoptosis and ferroptosis rates are significantly decreased in vitro (P<0.05).
CONCLUSIONS NDUFA13 is highly expressed in both human and mouse aortic aneurysm, and it has different expression profile in human hereditary vascular disease such as Marfan’s syndrome. After NDUFA13 knock out in smooth muscle cells, mouse aortic aneurysm is dramatically attenuated in both elastase and BAPN induced model. The in-vitro study of MOVAS and SMC in programmed cell death has further elucidated its protection role for vascular injury after knockdown of NDUFA13. Our data indicates that NDUFA13 might be a promising therapeutic target for vascular injury.
GW31-e1298
Jinyong Huang1, Tianming Teng1, Yuchen Xue1, Longfei Huang1, Zhelong Xu2, Yuemin Sun1
1Tianjin Medical University General Hospital
2Tianjin Medical University
OBJECTIVES This study evaluated the correlation between Zip14 and expression of some proteins of endoplasmic reticulum stress (ERS) in hypertrophied hearts of rats.
METHODS Dahl salt-sensitive rats were fed a high salt diet to establish a left ventricular hypertrophy (LVH) rat model.
RESULTS Compared with the Control group, Zip14 mRNA and protein expression levels in the LVH rat heart were markedly increased (P<0.01). The zinc content in rat heart tissue was significantly increased in the LVH group compared with the Control group (P<0.05). Activating transcription factor (ATF)4, ATF6, and x box-binding protein 1 (xBP1) mRNA levels were increased in the LVH rat heart compared with Control hearts (P<0.001). Compared with the control group, C/EBP homologous protein (CHOP) and immunoglobulin-binding protein (BiP) mRNA and protein levels were markedly increased in the LVH rat heart (P<0.05, P<0.01). Linear regression models showed that Zip14 mRNA expression levels were positively correlated with zinc concentration, ATF4 and ATF6 mRNA expression levels in Control hearts (P=0.0005, P=0.0052 and P=0.0026, respectively) and LVH rat hearts (P<0.0001, P=0.0119 and P=0.0033, respectively).
CONCLUSIONS In summary, the upregulation of Zip14 in LVH rat hearts correlated with zinc accumulation and the induction of ERS.
GW31-e1326
Xuejun Wang, Lingmei Qian
The First Affiliated Hospital of Nanjing Medical University
OBJECTIVES Doxorubicin is served as a common anti-tumor drugs, however the application is limited by its cardiotoxicity. Circular RNAs (circRNAs) have been proved to be associated with cardiovascular diseases in previous studies. However, the role of circRNAs on DOX-induced cardiotoxicity still needs to be clarified. This research is to investigate the differential expression mode of circular RNAs (circRNAs) in mice cardiomyocytes during doxorubicin induced cardiotoxicity.
METHODS Two groups of mice were injected with equal amount of normal saline and doxorubicin, the isolated mice heart tissues were submitted to next generation RNA-sequencing. Expression profiles of circRNAs and constructed circRNA-miRNA-mRNA networks were analyzed. Overall 48 upregulated circRNAs and 16 downregulated circRNAs in cardiotoxicity apoptosis were assessed. Bioinformatics analysis revealed several potential biological pathways might be related to cardiac toxicity induced by DOX. CircArhgap12 was up-regulated in the DOX treated group.
RESULTS The silencing of CircArhgap12 significantly attenuated DOX-induced apoptosis and oxidative stress. Overexpression of miR-135a-5p in rat primary cardiomyocytes reduced the anti-apoptotic effect of si-circArhgap12 and accelerated oxidative stress. With bioinformatic analysis of miR-135a-5p, it might have a potential target site for ADCY1 mRNA.
CONCLUSIONS Our research demonstrated that expression profiles of circRNAs were significantly modified and circArhgap12 might play a competitive function among endogenous RNAs in DOX-induced cardiotoxicity, which shed a new light on the mechanism of ceRNA network in DOX-induced cardiotoxicity.
GW31-e1330
Jing Lou, Jie Wu, Xing Zhang, Feng Gao
Air Force Medical University
OBJECTIVES The significance of exercise in prevention and treatment of cardiovascular diseases is receiving increasing attention. Cardiovascular protective effects of exercise largely depend on its effect in improving blood perfusion and increasing vascular density of organs. miRNA has well-established functions in cancer, metabolic disorders, and cardiovascular diseases and it can travel around body contained within exosome, but the mechanism of exercise-induced angiogenesis by miRNA has not been fully elucidated. In this research, we are aim to identify the specific miRNA that cause angiogenesis during exercise and elucidate the mechanism underlying specific miRNA induced angiogenesis and explore the application potential of the selected miRNA as a pro-angiogenic drug.
METHODS We established aerobic exercise model with a combination of 3-day adaptive training and 9-day treadmill training. AgomiR was adopted to up-regulate the selected miRNA by skeletal muscle point injection, and the AntagomiR was injected through tail vein to interfere with the circulating miRNA level. To determine whether liver derived specific miRNA contributed to exercise-induced angiogenesis, mice were injected with AAV8 through tail vein 1 month before treadmill training, which carried specific antisense sequence. Wound scratch, tube formation and aortic ring assay were used to estimate angiogenesis effect in vitro. Wound healing and matrigel plug assays were used to detect angiogenesis in vivo.
RESULTS Nine days treadmill training significantly increased the expression of VEGF and CD31 in the quadriceps muscle of mice. According to cell proliferation assay, we screened miR-122-5p which was significantly promoted endothelial cell proliferation. Endothelial cell migration, tube formation and aortic ring assay confirmed that miR-122-5p had pro-angiogenic capacity. When we point injected AgomiR-122-5p in skeletal muscle, WB and IF results showed that the protein levels of VEGF and CD31 as well as the CD31 fluorescence density were increased, suggested that miR-122-5p can increase local vascular density in vivo. Then we injected AntagomiR-122-5p or AAV(8)-Anti-122-5p in tail vein to decrease circulatory or liver-derived miR-122-5p, the WB and IF results showed that the VEGF and CD31 expression increased in skeletal muscle of exercise group compared with the sedentary group, while it was not significantly different after AntagomiR-122-5p or AAV(8)-Anti-122-5p injection between the exercise group and the sedentary group, suggested that liver-derived miR-122-5p play a key role in exercise-induced angiogenesis. We used website to predicted and screened the downstream target and found Agpat1 (an enzyme participate triglyceride biosynthesis) is the key gene for this miRNA. CPT1A & CD36 were increased and Agpat1 was decrease when miR-122-5p was promoted in vitro or in vivo. We found that aerobic exercise can effectively increase lipid metabolism through miR-122-5p to promote angiogenesis.
CONCLUSIONS Our research found miR-122-5p as an angiogenic factor. miR-122-5p promoted angiogenesis by up-regulating the expression of CD36 & CPT1A and down-regulating the key target gene Agpat1, which in turn increases endothelial cell fatty acid utilization and VEGF expression. The exercised-induced angiogenesis partially depends on the elevation of circulating miR-122-5p, indicating that miR-122-5p may work as an exerkine and potentially help with the pro-angiogenesis and wound healing treatment.
GW31-e1331
Mengqi Su, Yue Li
The First Affiliated Hospital, Harbin Medical University
OBJECTIVES we noticed that adipocytes from the white adipose tissue of patients with HUA were hypertrophied and had decreased UCP1 expression. To test the effects of UA on adipose tissue.
METHODS we built both in vitro and in vivo HUA models and elucidated that a high level of UA could induce hypertrophy of adipocytes, inhibit their hyperplasia and reduce their beige-like characteristics. According to mRNA-sequencing analysis, UA significantly decreased the expression of leptin in adipocytes, which was closely related to fatty acid metabolism and the AMPK signalling pathway, as indicated by KEGG pathway analysis.
RESULTS Our observations confirm that UA is involved in the aetiology of metabolic abnormalities in adipose tissue by regulating leptin-AMPK pathway, and metformin could lessen HUA-induced serum FFA elevation and insulin resistance by improving adipose tissue function via AMPK activation.
CONCLUSIONS metformin could represent a novel treatment strategy for HUA-related metabolic disorders.
GW31-e1339
Yu Kang1, Xiaojing Chen1, Qiaowei Chen1, Mian Wang1, Mingqiang Rong2, Qing Zhang1
1Department of Cardiology, West China Hospital, Sichuan University
2The National & Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha, Hunan, China
OBJECTIVES To explore the phenomenon of mitral leaflet growth and its relationship to the dynamic change of ischemic mitral regurgitation (IMR) in rhesus monkey by using transthoracic echocardiography (TTE) and proteomic methods.
METHODS Rhesus monkeys who had completed left circumflex artery occlusion surgery received TTE at baseline, 1-, 2-, 3-, 4-, 8- and 16-week after surgery. Days within one week post-surgery was defined as the acute phase, and after that to the 16-week post-surgery was the chronic phase. The global remodeling parameters included left ventricular end-diastolic volume (LVEDV), end-systolic volume (LVESV) and ejection fraction (LVEF). The parameters to evaluate mitral annular remodeling [mitral annular diameter plus tenting height (MAD+TH)] and the length of anterior mitral leaflet (AML) and the posterior mitral leaflet (PML) were measured at the parasternal left ventricular long axis view. The IMR severity were measured as well. All monkeys who survived at 16-week post-surgery were sacrificed to obtain samples of mitral leaflets. The corresponding tissues of 2 normal monkeys were taken as the control. Differential protein analysis, gene enrichment analysis, functional annotation and pathway and network analysis were performed.
RESULTS A total of 13 monkey survived for 16 weeks after surgery. In the acute phase, 10 monkeys developed IMR (mild IMR, n=5; moderate IMR, n=5) which were defined as IMR (+) group. In IMR (+) group, LVESV (8.7±2.5 vs. 12.1±4.4 mL, P=0.006), (MAD+TH) (16.2±1.3 vs. 18.4±2.4 mm, P=0.003) and TA (0.22±0.04 vs. 0.38±0.09 cm2, P=0.001) were significantly increased, but LVEF (59.1±4.9 vs. 48.4±11.5%, P=0.006) decreased when compared with baseline. PML slightly increased (6.3±0.8 vs. 6.9±0.6 mm, P=0.024), but the AML did not change (P=0.052) (Table 1). During the chronic phase, there were 8 monkeys in the IMR (+) group with stable/reduced IMR, but their LVEDV (P=0.064), LVESV (P=0.054) and LVEF (P=0.265) showed no statistically difference at each time point during the chronic period. However, (MAD+TH) increased further (1- vs. 4- vs. 16-week 16: 18.4±2.4 vs. 20.7±3.5 vs. 19.1±3.4, P=0.009). AML (1- vs. 4- vs. 16-week: 14.7±1.7 vs. 15.8±2.3 vs. 15.7±2.2, P=0.001) and PML (1- vs. 4- vs. 16-week: 6.9±0.6 vs. 7.9±1.0 vs. 8.0±1.3, P=0.013) increased correspondingly (Table 2). By protemetic analysis, 118 proteins were up-regulated and 52 proteins were down-regulated in AML, most of these differential expressed proteins riched in cellular mitochondrial components. The number of up-regulated proteins was 50, and the number of the down-regulated proteins was 10 in PML, most of these differential expressed proteins riched in extracellular matrix components.
CONCLUSIONS The growth of mitral leaflets failed to adapt to the structural and functional changes of LV and mitral apparatus in the acute phase might be one of the mechanisms of acute IMR. The further growth of mitral leaflets adapted to the degree of cardiac remodeling might be the reason for the IMR reduction in the chronic phase. This study further confirmed the existence of the differential expression of protein of the mitral leaflet in the rhesus monkey IMR model. Further study will be helpful to the explore the growth mechanism behind mitral leaflet growth and to develop new therapeutic targets.
GW31-e1341
Qinchao Wu, Baochen Bai, Xianming Chu
Department of Cardiology, The Affiliated hospital of Qingdao University
OBJECTIVES In recent years, there has been a revolutionary decrease in cancer-related mortality and prolonged survival due to the induction of novel targeted drugs. Nevertheless, targeted therapies also bring up inevitable adverse events and there is a lack of precise mechanisms and therapeutic measures about novel targeted agents. This review focuses on the molecular mechanisms of cardiotoxicity induced by some targeted agents. The purpose of this article is to stimulate awareness of the emerging cardiotoxicity of novel targeted anticancer drugs and provide novel insights into cardiovascular disease.
METHODS “Cardiotoxicity”, “targeted drugs”, “HER2”, “trastuzumab”, “angiogenesis inhibitor”, “VEGF inhibitor” and “tyrosine kinase inhibitors” were used as keywords for article searches. We reviewed related articles and presented updated knowledge of the potential mechanisms of cardiotoxicity induced by targeted drugs.
RESULTS HER2-targeting drugs significantly increases the risk of heart failure. The mechanisms are involved in the inhibition of HER-2 and downstream signaling pathways including PI3K/Akt, MEK/Erk and Src/FAK pathways. Block of the downstream effectors result in mitochondrial dysfunction and cell injury, which predisposes to the development to heart failure. In addition, bevacizumab, an angiogenesis inhibitor, brings up inevitable adverse events, particularly hypertension. The potential mechanisms are related to the inhibition of the PI3K/Akt pathway and a decreased production of nitric oxide, prostacyclin, and increased level of endothelin-1, which promotes vasoconstriction, resulting in endothelium dysfunction. Besides, some tyrosine kinase inhibitors including sunitinib, sorafenib, and vandetanib are associated with higher incidence of QT interval prolongation. The inhibition of PI3K/Akt activity leads to disorders of related ion channels, which prolongs ventricular repolarization and gives rise to a prolonged QT interval.
CONCLUSIONS In this review, we concentrate on the molecular mechanisms of cardiotoxicity induced by certain targeted drugs and find several important signal pathways including PI3K-Akt and MEK-Erk signaling, as well as some potential mechanisms. Notably, the PI3K-Akt pathway exerts complicated biological effects by increasing NO production via activation of eNOS, inhibiting apoptosis, and modulating multiple ion channels. Currently, the management of cardiotoxicity induced by antitumor agents mainly depends on early detection, careful monitoring, and empirical treatment, there is still a lack of specific recommendations by guidelines. Future treatments for targeted drug-induced cardiotoxicity depend on a better understanding of mechanisms and further clinical evidence.
GW31-e1349
Jiang He, Yumin Qiu, Bin Dong, Bingbo Yu, Zhe Zhou, Zhichao Wang, Wenhao Xia
Department of Hypertension and Vascular Disease, The First Affiliated Hospital, Sun Yat-sen University
OBJECTIVES Decline in angiogenic capacity is the leading cause of age-related cardiovascular disease. Mitochondrial oxidative stress is implicated in the abnormalities of vascular formation. Sirtuin 5 (SIRT5) plays a critical role in maintenance of mitochondrial redox homeostasis through regulating the succinylome; however, the exact function of SIRT5 in angiogenesis and upstream regulated mechanisms remain largely unknown. The present study aim to investigate the role of microRNAs in SIRT5-mediated mitochondrial oxidative stress via desuccinylation of antioxidant enzymes.
METHODS Cellular signaling was analyzed in primary aortic endothelial cells (PAEC). miRNA array was used to identify differential expression of miRNAs. Electron spin resonance was used to detect mitochondrial superoxide anion (O2.- ) generation. Proteomic analysis, luciferase assays, immunoprecipitation were performed.
RESULTS Duplicate senescence in PAEC was associated with decreased angiogenic capacity and excessive mitochondrial O2.- production. miR-19b was highly enriched in senescent PAEC. Induction of miR-19b in PAEC impeded the formation of normal vascular structures along with downregulation of SIRT5 and global protein succinylation. Proteomic analysis of senescent PAEC revealed the specific lysine succinylation of a central mitochondrial antioxidant enzyme, SOD1. miR-19b downregulated SIRT5 expression through direct 3′-untranslated region targeting. SIRT5 provoked desuccinylation of SOD1 associated with enhanced enzymatic activity, leading to the elimination of O2.- . Treatments with miR-19b mimics or SIRT5 silencing augmented mitochondrial oxidative damage with subsequent inhibition of angiogenesis. Suppression of miR-19b with antimiR oligonucleotides attenuated mitochondrial oxidative damage and rescued senescence-related abnormity in vascular formation via SIRT5-mediated SOD1 Desuccinylation.
CONCLUSIONS We demonstrated for the first time that miR-19b involves mitochondrial oxidative stress via direct suppression of sirt5 and enzymatic inhibition of SOD1. Blockage of MiR-19b may be a novel therapeutic target for age-related disorder of vascular formation.
GW31-e1352
Chunyan Yang, Ping Yang
China-Japan Union Hospital, Jilin University, Changchun, China
OBJECTIVES Ghrelin, a novel growth hormone-releasing peptide, potentially improves cardiac function, but the mechanisms remain unclear. Angiotensin II (Ang II), an important neurohormonal factor during heart failure, can induce myocardial fibrosis. However, the molecular mechanism of Ang II inducing myocardial fibrosis remains unclear, especially its role and mechanism in the progression of myocardial fibrosis in heart failure after myocardial infarction (MI). Galactosin-3 (Gal-3) is a member of the galactin family and is closely related to myocardial fibrosis. We hypothesized that Ang II may induce Gal-3 expression through its type 1 receptor (AT1R), thereby promoting myocardial fibrosis of heart failure after myocardial infarction, while Ghrelin can inhibit the above useful effect.
METHODS In the study, the left anterior descending coronary artery was ligated to establish a rat model of heart failure (HF), and then treated with ghrelin (100 μg/kg, subcutaneous injection, bid); the cardiac fibroblasts from neonatal rats were cultured and stimulated with Ang II (0.1 μM) and ghrelin (0.1 μM) to explore the role and mechanism of ghrelin in myocardial fibrosis. Hemodynamic and serum brain natriuretic peptide (BNP) concentrations were measured to assess cardiac function. Left ventricular mass index (LVMI), hematoxylin and eosin (H&E) staining, and Masson’s trichrome staining were performed to evaluate myocardial fibrosis. To further explore whether ghrelin inhibits myocardial fibrosis by inhibiting Ang II/AT1R/Gal-3 axis, the levels of AT1R and Gal-3 were examined by immunohistochemistry, real-time quantitative PCR and ELISA in vivo and in vitro. Finally, to investigate the effect of Gal-3 on myocardial fibrosis, the primary rat cardiac fibroblasts (CFs) were cultured and treated with Gal-3, the proliferation of CFs was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Also, the expressions of type I and type III collagen (Col I and Col III) were measured by real-time quantitative PCR.
RESULTS The results showed that ghrelin significantly improved cardiac function and hemodynamics in HF rats after myocardial infarction (MI), and inhibited type I and type III collagen expression in vivo; inhibited the CFs proliferation and type I and type III collagen expression induced by Ang II in vitro. The results also showed that Gal-3 levels were significantly increased in the MI group, and ghrelin administered downregulated Gal-3 expression. Furthermore, ghrelin decreased Ang II-induced Gal-3 expression in primary rat cardiomyocytes in vitro by downregulating AT1R expression. In addition, Ang II upregulated gal-3 expression through AT1R, and Gal-3 treated cultured primary rat cardiac fibroblasts (CFs) showed increased proliferation and enhanced expressions of type I and type III collagen.
CONCLUSIONS These data suggest that Ang II can induce Gal-3 expression through AT1R, thereby promoting the proliferation of CFs and the expression of type I and III collagen to promote HF progression, while Ghrelin can inhibit myocardial fibrosis and protect heart failure by inhibiting Ang II/AT1R/Gal-3 axis.
GW31-e1366
Alex Chia Yu Chang
Department of Cardiology and Shanghai Institute of Precision Medicine, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine
OBJECTIVES Dilated Cardiomyopathy (DCM) is a cardiac disease characterized by dilatation of the ventricular chamber resulting in decreased systolic function. DCM mostly occurs between the ages of 20–60 affecting approximately 1 in 2500. Between 30–50% of DCM cases are inherited with ~33 known DCM mutations classified as genetic DCM. DCM is the most common cause of heart failure after coronary artery disease and hypertension, as well as the leading indication for heart transplantations. Initially, therapy includes medications such as ACE inhibitors, angiotensin receptor antagonists, beta-blockers, aldosterone antagonists, digoxin, and diuretics. Some patients require surgery to have a pacemaker or defibrillator placed. For patients whose symptoms do not respond to these treatments, waiting for a heart transplant is the only option. We recently observed that telomeres, DNA-repeat sequences that protect the ends of chromosomes, are reduced in length by >40% in cardiomyocytes of genetic DCM patient hearts lacking dystrophin, Troponin t2, or Titin. We seek to test if telomere shortening is a hallmark of genetic cardiomyopathy.
METHODS Typically, telomeres shorten at every cell division due to replication insufficiency and are markers of cellular aging, but in healthy postnatal cardiomyocytes, telomere lengths are maintained throughout life, presumably because these cells do not proliferate. Here we used human induced pluripotent stem cell derived cardiomyocytes as model to study the mechanism of telomere shortening.
RESULTS This unexpected telomere shortening is recapitulated in cardiomyocytes differentiated from human induced pluripotent stem cells (hiPSC-CM) derived from DCM patients, enabling mechanistic studies. We observed aberrant calcium handling and decreased contractility using bioengineered micropatterned hydrogel traction force microscopy. Here we present new evidence where aberrant contraction results in telomere deprotection and resection in in non-dividing hiPSC-CMs. Induction of DNA damage response culminated in mitochondrial dysfunction and apoptosis.
CONCLUSIONS The proposed model enables the study of cause and effect and tests of interventions.
GW31-e1367
Honghui Wang1, Dongjiu Li2, Chia Yu Alex Chang1,2
1Department of Cardiology, Shanghai Institute of Precision Medicine, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, China
2Department of Cardiology, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, China
OBJECTIVES To study whether there is an accelerated aging (LTL shortening and increased inflammatory response) in the younger population that leads to higher incidence of CHD.
METHODS Blood samples and clinical data of more than 2000 patients with CHD undergoing coronary angiography or PCI in the department of cardiology of Shanghai ninth people’s hospital from 2016 to 2019 were analyzed. Based on age inter quartile range (IQR) of all patients, CAG and PCI patients were divided into four age groups: <60 y, 60–65 y, 66–73 y and ≥73 y. LTL and clinical related factors in each group were measured and analyzed.
RESULTS Compared with CAG patients, PCI patients exhibited significantly shorter LTL. Interestingly, shorter LTL did not result in decrease in mitochondrial copy number. Importantly, <60 y PCI patients showed significant decrease in LTL similar to older PCI patients.
CONCLUSIONS These results suggest that LTL shortening is strongly associated with more severe CHD that required PCI treatment. Moreover, regardless of age, PCI patients overall exhibited shorter LTL compared to CAG cohort and provides the necessary samples to further characterize and define molecular aging in CHD.
GW31-e1372
Yangyang Liu1,2, Felix Jansen2
1Department of Internal Medicine Cardiac Disease, Shanghai Ninth People’s Hospital, Shanghai JiaoTong University School of Medicine
2Heart Center Bonn, Department of Internal Medicine II, University Hospital Bonn, University of Bonn Venusberg-Campus 1, 53127 Bonn, Germany
OBJECTIVES Extracellular vesicle (EV)-incorporated long noncoding RNAs (lncRNAs) have emerged as biomarkers and regulators of cardiovascular disease. Our aim was to study the expression pattern of circulating small EV (sEV)-incorporated lncRNAs in patients with and without coronary artery disease (CAD).
METHODS PCR-based human lncRNA array analysis revealed that certain sEV-lncRNAs are significantly different in patients with CAD compared to patients without CAD. Four atherosclerosis-related lncRNAs (PUNISHER, GAS5, MALAT1, and H19) were quantified in the circulating sEV by using real-time quantitative PCR (RT-qPCR) in 60 patients with (n=30) or without (n=30) CAD.
RESULTS Four atherosclerosis-related lncRNAs (PUNISHER, GAS5, MALAT1, and H19) were quantified in the circulating sEV by using real-time quantitative PCR (RT-qPCR) in 60 patients with (n=30) or without (n=30) CAD. Among these, PUNISHER (P=0.002) and GAS5 (0.02) were significantly increased in patients with CAD. In vitro, atherosclerotic stimuli upregulated PUNISHER levels in endothelial cells (EC) and in the corresponding sEV. Labeling of sEV and RT-qPCR demonstrated the transportation of PUNISHER into recipient ECs, which accelerated cell migration, proliferation, and tube formation. Mechanistically, the RNA-binding protein hnRNPK was identified to regulate PUNISHER loading into sEV. Knockdown of PUNISHER abrogated the sEV-mediated effects on EC migration, proliferation, and tube formation. PCR-based gene profiling showed that the expression of VEGFA RNA was increased in ECs by sEV treatments. Knockdown of PUNISHER in sEV abrogated the EV-mediated promotion of VEGFA gene- and protein expression.
CONCLUSIONS The circulating lncRNA PUNISHER is increased in CAD patients. Intercellular transfer of sEV-incorporated PUNISHER promotes a pro-angiogenic phenotype via a VEGFA-dependent mechanism.
GW31-e1377
Hui Zhang1, Liuzhi Hao2, JunFeng Zhang1, Changqian Wang1, Aizheng Chen2, Huili Zhang1
1Department of Cardiology, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, PR China
2Fujian Provincial Key Laboratory of Biochemical Technology, Institute of Biomaterials and Tissue Engineering, Huaqiao University, Xiamen 361021, PR China
OBJECTIVES Pulmonary drug delivery offers tremendous opportunity, both systemically and locally, for treating pulmonary disorders. Hydrogen sulfide (H2S) has recently emerged as a novel gaseous mediator with protective actions in treating pulmonary arterial hypertension (PAH). However, the therapeutic potential of H2S has been substantially hampered due to the lack of appropriate donors that could mimic the tightly controlled endogenous production in response to specific biological conditions. Therefore, an innovative formulation based on inhalable porous poly(lactic-co-glycolic acid) (PLGA) microspheres containing H2S-releasing aspirin derivative (ACS14) (ACS14 MSs) is fabricated using the microfluidic technology. In addition to physicochemical attributes, cytotoxicity, lung deposition characteristics, and H2S release profile, the protective effect of ACS14 MSs on PAH and the underlying mechanism concerning the process of endothelial-to-mesenchymal transition (EndMT) are evaluated both in vitro and in vivo.
METHODS ACS14 MSs were fabricated using microfluidic technology. The characterizations of ACS14 MSs including the surface morphology, the aerodynamic properties, the chemical functionalities, the crystal characteristics, the thermal properties were analyzed. Drug loading and encapsulation efficiencies were measured by HPLC. The cytotoxicity were analyzed by CCK-8 cell viability detection, TUNEL staining and Annexin V/PI flow cytometry. The lung deposition characteristics were detected using in vivo fluorescence imaging. Eight-week-old, male SD rats (200±20 g) were randomly given monocrotaline (60 mg/kg, i.p.) or saline. Seven days after the injection of MCT, rats were daily administered with sildenafil (25 mg/kg, i.g.), ACS14 MSs (46.5 mg/kg, i.t.), PLAG MSs (46.5 mg/kg, i.t.), or ACS14 (12 mg/kg, i.p.) daily for 14 days. Pulmonary artery pressure and the right-heart function were observed 21 days after MCT injection. Pulmonary arterial remodeling was examined by HE staining. EndMT was investigated by western blot immunoblotting and immunofluorescence. NFκB p65 DNA binding activity was also assayed. HPAECs were pre-incubated with PLGA MSs (50 μM, 2 h), sildenafil (1 μM, 30 min), ACS14 (25 μM or 50 μM, 2 h) or ACS14 MSs (25 μM or 50 μM, 2 h) and then stimulated with TGF-β1 (10 ng/mL, 1 h, 3 d or 10 d) to evaluate morphology changes, ultrastructural transformation in HPAECs, the process of EndMT and its underlying mechanism.
RESULTS These porous ACS14 MSs displayed excellent aerodynamic properties, high drug loading, and entrapment efficiency with slow drug release behavior and negligible cytotoxicity, indicating their suitability for delivering ACS14 via pulmonary administration. In human pulmonary artery endothelial cells (HPAECs), ACS14 MSs inhibited TGF-β1-induced EndMT, an important process in vascular remodeling of PAH and the activation of NF-κB-Snail pathway in a dose-dependent manner. Moreover, in a rat model of monocrotaline-induced PAH, daily inhalation of ACS14 MSs significantly inhibited the process of EndMT by suppressing the induction of NF-κB-Snail pathway and thus improved the severity of PAH with comparable efficacy to oral administration with sildenafil, a conventional PAH treatment.
CONCLUSIONS In conclusion, our findings demonstrated that the designed microfluidics-assisted porous ACS14 MSs have shown great potential for the inhalation treatment of PAH.
GW31-e1378
Jianan Pan, Hao Lin, Jianying Yu, Huili Zhang, Junfeng Zhang, Changqian Wang, Jun Gu
Department of Cardiology, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, People’s Republic of China
OBJECTIVES A relationship between the abundance of epicardial adipose tissue (EAT) and the risk of atrial fibrillation (AF) in diabetes mellitus (DM) has been reported. And browning of EAT might be a novel approach for prevention or treatment in AF by adjusting atrial fibrosis. MicroRNA-21 (miR-21) have shown been a regulatory factor in atrial fibrosis. The aim of this study was to examine the role of miR-21-3p in EAT browning in hyperglycemia conditions.
METHODS In vivo, Normal C57BL/6 wild type (WT) and miR-21 knockout (KO) mice were used to establish the diabetic model by intraperitoneal injection of streptozotocin (STZ). In vitro, The EAT adipocytes from miR-21 KO mice were cultured and transfected with miR-21-3p mimic or miR-21-5p mimic in both HG or LG conditions. The browning of EAT were assessed by western blotting and immunofluorescence, and the release of inflammatory factors were assessed by ELISA. The gain- and loss-of-function experiments were used to identified fibroblast growth factor receptor 1 (FGFR1) as the target gene of miR-21-3p, and the regulatory pathway of miR-21-3p FGFR1, fibroblast growth factor 21 (FGF21) and peroxisome proliferator-activated receptor gamma (PPARγ) that controlled EAT browning under hyperglycemia conditions.
RESULTS MiR-21 KO clearly ameliorated the atrial fibrosis in the diabetic mice. miR-21-3p as a key regulator that controls EAT browning and participates in atrial fibrosis under hyperglycemia conditions. Moreover, our gain- and loss-of-function experiments showed that FGFR1, as a direct target of miR-21-3p identified a regulatory pathway in EAT adipocytes consisting of miR-21-3p, FGFR1, FGF21 and PPARγ.
CONCLUSIONS MiR-21-3p regulated EAT browning and the inflammatory factors releasing i under hyperglycemia conditions by targeting FGFR1/FGF21/PPARγ pathway.
TRANSLATIONAL RESEARCH OF CARDIOVASCULAR DISEASE
GW31-e0030
Chuannan Zhai1,2, Hongliang Cong1,2
1NanKai University, School of Medicine
2Tianjin Chest Hospital, Department of Cardiology
OBJECTIVES Recent studies have shown that blood-based miRNAs are dysregulated in patients with AMI and are therefore a potential tool for the diagnosis of AMI. However, the diagnostic value of miRNAs is still inconsistent due to differing results among previous studies. Therefore, this study summarized and evaluated studies focused on microRNAs as novel biomarkers for the diagnosis of AMI from the last ten years.
METHODS MEDLINE (including PubMed), the Cochrane Central database, and EMBASE were searched between January 2010 and December 2019. Studies that assessed the diagnosis accuracy of circulating microRNAs in AMI were chosen. The number of significantly dysregulated miRNAs identified, patients demographics, type of clinical sample, method of miRNA detection, and type of normalization were included. The pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio, and area under the curve were used to assess the overall test performance of miRNAs.
RESULTS A total of 58 studies that included 8206 participants assessed the diagnosis accuracy of circulating miRNAs in AMI. The main results of the meta-analyses are as follows: Total miRNAs: the overall pooled sensitivity and specificity were 0.82 (95% CI: 0.79–0.85) and 0.87 (95% CI: 0.84–0.90), respectively. The AUC value was 0.91 (95% CI: 0.88–0.93) in the overall summary receiver operator characteristic (SROC) curve. Results from the panel of two miRNAs: sensitivity: 0.88 (0.77–0.94), specificity: 0.84 (0.72–0.91), AUC: 0.92 (0.90–0.94); The panel of three miRNAs: sensitivity: 0.91 (0.85–0.94); specificity: 0.87 (0.77–0.92), AUC: 0.92 (0.89–0.94); miRNA-1: sensitivity: 0.78 (0.71–0.84), specificity: 0.86 (0.77–0.91), AUC: 0.88 (0.85–0.90); miRNA-133a: sensitivity: 0.85 (0.69–0.94), specificity: 0.92 (0.61–0.99), AUC: 0.93 (0.91–0.95); miRNA-208b: sensitivity: 0.80 (0.69–0.88), specificity: 0.96 (0.77–0.99), AUC: 0.91 (0.88–0.93); miRNA-499: sensitivity: 0.80 (0.69–0.88), specificity: 0.96 (0.77–0.99), AUC: 0.91 (0.88–0.93).
CONCLUSIONS MiRNAs may be used as potential biomarkers for the detection of AMI. For single, stand-alone miRNAs, miRNA-499 may have better diagnostic accuracy compared to other miRNAs. We propose that a panel of multiple miRNAs with high sensitivity and specificity should be tested.
GW31-e0041
Xingxing Cai1, Yuli Yang1, Wei Wang1, Yichen Shen1, Li Qian2, Zhixing Wei1, Taizhong Chen1, Jing Cai1, Runmin Chi1, Shunxuan Yu1, Keke Li1, Mawei Jiang1, Yigang Li1
1Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine
2Affiliated Hospital of Nantong University
OBJECTIVES Noninvasive stereotactic body radiotherapy (SBRT) is a novel technique for renal denervation (RDN). The purpose of this study was to investigate the chronological evolution and mechanism of SBRT-induced renal nerve injury, as well as its antihypertensive effect and safety within 6 months in a hypertensive swine model.
METHODS Bama swine were randomly divided into 1-month group (n=4), 3-month group (n=4), 6-month group (n=5) according to the duration of follow-up post-RDN with a single dose of 25 Gy delivered by SBRT, and additional 5 swine served as control. Hypertension was induced by subcutaneous implantation of deoxycorticosterone acetate (DOCA) pellets in combination with a high-salt diet. Blood pressure and renal function were measured at baseline, 1 month, 3 months and 6 months after treatment. Abdominal contrast-enhanced CT scan was performed at the predetermined endpoint. The concentration of renal norepinephrine was measured by high performance liquid chromatography-mass spectrometry. The characteristics of injured nerves were analyzed by standard semi-quantitative scoring method. Nerve apoptosis was determined by TUNEL and immunofluorescence. Function of sympathetic nerves and nerve regeneration were evaluated with immunostains against tyrosine hydroxylase (TH) and growth-associated protein 43 (GAP43).
RESULTS All animals survived to the predetermined endpoint. Blood pressure at baseline, 1 month, 3 months and 6 months post-RDN were comparable to control, whereas renal norepinephrine was significantly lower at 6 months (384.2±103.6 ng/g vs. 700.9±121.8 ng/g, P<0.05). Compared with control (1.4±0.4), nerve injury score was greatest at 6 months (3.2±0.8), followed by 3 months (2.5±0.4) and 1 month (2.3±0.3) (P<0.001). The proportion of TUNEL positive nerves at 3 months and 6 months were both significantly higher than control (14.2±3.3%, 23.0±7.3% vs. 1.3±2.5%, P<0.05). Compared with 1 month, arteriolar injury score was significantly greater at 3 and 6 months (2.5±0.4, 2.4±0.2 vs. 1.6±0.6, P<0.05), and arteriolar occlusion due to intimal hyperplasia and thrombosis was observed at 3 months and more common at 6 months. Although there seems to be a trend showing more severe soft tissue injury characterized by denatured collagen, fat necrosis and fibrosis at 3 and 6 months, no statistical difference was found between groups (3.0±0.8, 3.1±0.2 vs. 2.0±1.4, P>0.05). Compared with control, TH scores were significantly lower (1.6±0.5, 1.4±0.5, 1.6±0.5 vs. 2.6±0.5, P<0.05) and GAP43 scores were similar (1.8±0.4, 2.0±0.0, 1.6±0.5 vs. 1.6±0.5, P>0.05) after RDN. CT images and renal function remained normal, notwithstanding 1 apparent focal pathological renal injury at 6 months.
CONCLUSIONS SBRT delivering 25 Gy for RDN maintained a promising safety profile with sustainable reduction of sympathetic activity up to 6 months, despite failure to lower BP in the DOCA-salt hypertensive swine model. Radiation-induced renal nerve injury aggravated over time, resulting from direct effects of irradiation and secondary effects of regional fibrosis and damaged microvasculature which induced nerve apoptosis and necrosis, as well as inhibition of nerve regeneration.
GW31-e0141
Chuannan Zhai1,2, Hongliang Cong1,2
1School of Medicine, NanKai University, Tianjin, China
2Tianjin Chest Hospital, Tianjin, China
OBJECTIVES MicroRNAs (miRNAs) have been shown to play a critical role in the development of and morbidity from cardiovascular diseases. However, their value as novel prognostic biomarkers in coronary artery disease (CAD) remains unclear. The current study evaluated the value of circulating miRNAs in the prognosis of CAD over the past 10 years.
METHODS MEDLINE (including PubMed), Embase and the Cochrane Central database were searched from January 2010 to April 2020. Studies that assessed the prognosis values of circulating microRNAs in CAD were chosen. A meta-analysis was conducted to evaluate the correlation between circulating miRNAs expressions and mortality, major adverse cardiovascular events (MACEs), and left ventricular adverse remodeling (LVAR) among these studies.
RESULTS Twenty studies involving 4513 patients were included in this meta-analysis. Total miRNAs were associated with high mortality (combined HR: 1.77, 95% CI: 1.47–2.13), MACEs (combined HR: 1.22, 95% CI: 1.03–1.45; combined OR: 1.80, 95% CI: 1.47–2.20), and LVAR (combined OR: 1.18, 95% CI: 1.12–1.26) in patients with CAD. An elevated level of miRNA-208b was associated with high mortality (pooled HR: 2.55, 95% CI: 1.36–4.78) and MACEs (pooled OR: 1.80, 95% CI: 1.32–2.45). MiRNA-133a (pooled HR: 1.31, 95% CI: 0.93–1.83) might be related to the occurrence of MACEs. MiRNA-150 (pooled OR: 1.22, 95% CI: 1.12–1.34) was correlated with LVAR after acute myocardial infarction (AMI).
CONCLUSIONS The present study showed that circulating miRNAs may be valuable predictors of clinical outcomes in CAD. miRNA-133a, miRNA-208b, and miRNA-150 show potential as useful prognostic biomarkers in the follow-up of patients with CAD.
GW31-e0397
WenHui Wu1,2, Geraldine Vitry2, Valerie Nadeau2, Junichi Omura2, Mark Orcholski2, David Marsolais2, Eve Tremblay2, Sandra Martineau2, Sandra Breuils Bonnet2, Roxane Paulin2, Steeve Provencher2, Olivier Boucherat2, Sebastien Bonnet2
1Department of Cardio-Pulmonary Circulation, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai 200433, China
2Pulmonary Hypertension research Group, IUCPQ Research Centre, 2725, Chemin Sainte-Foy, QC, Canada G1V 4G5
OBJECTIVES Pulmonary fibrosis (PF) is an important commonality in pathogenesis of interstitial lung diseases, and pulmonary hypertension (PH) is a well-recognized severe complication of PF. Both significantly impact patients’ survival and functional status, but without validated treatments. In the lung, parenchymal and vascular remodeling share pathomechanisms, mainly including excessive proliferation and resistance to apoptosis of fibroblasts (LFs) and pulmonary arterial smooth muscle cells (PASMCs). In cancer, sustained cell proliferation is ensured, in part, by a fine tuning of cell cycle and DNA repair machinery, of which checkpoint kinases (CHK1 and CHK2) are critical regulators. They are upregulated in cancer and their inhibition are currently tested in clinical trials. However, these pathways have never been explored in PF. We thus hypothesized that CHK1/2 are upregulated in PF-PH and contribute to both fibrotic and vascular lesions in PF-PH patients.
METHODS Lung tissues, LFs and PASMCs were obtained from PF patients and health control. CHK1/2 and other protein expression levels were monitored by western blotting, and their location were confirmed by immunofluorescence. To study the effect of CHKs on HLFs and PASMCs proliferation and apoptosis in vitro, cultured human cells were exposed to 10% FBS or 0.1% FBS in presence of MK-8776, LY2606368 or vehicle for 48 hours.
RESULTS Increased DNA damage (γH2Ax and p(S4/S8)-RPA32) as well as augmented expression and activity of CHK1/2 was observed by WB in lung and isolated LFs from PF patients compared to controls (P<0.05, n=8–15 per group) with marked expression in fibroblastic foci. Similarly, increased levels of CHK1/2 and DNA damage were detected by WB and IF in remodeled pulmonary arteries (PA) of PF patients with or without PH. Similar findings were noted in mice exposed to bleomycin. In isolated PF-LFs and PASMCs, dual inhibition of CHK1/2 using MK-8776 and LY2606368 significantly reduced expression of TCTP and RAD51 (two factors required for efficient DNA repair), leading to exacerbation of DNA damage and resulted in reduced cell proliferation (Ki67 labeling, WB PCNA) and resistance to apoptosis (Annexin V assay, WB Survivin). Furthermore, inhibition of CHK1/2 mitigated the hyper-activated state of PF-LFs, as illustrated by reduced expression of FN, CTGF, and pSTAT3 (WB). Similar results were observed in control LFs exposed to TGF-β1; all P<0.05.
CONCLUSIONS Our data provide compelling evidence that CHK1/2 are involved in lung fibrogenesis and PA remodeling in PF. Current experiments aim to determine whether CHK1/2 inhibitors elicit beneficial effects in animal models of PF-PH.
GW31-e0456
Quanbin Dong1, Huihui Bao1, Liang Liu1, Lingyong Zhu2, Xiaoshu Cheng1
1The Second Affiliated Hospital of Nanchang University
2East China University of Science and Technology
OBJECTIVES Cardiovascular implantable electronic device infections are associated with high hospitalization, mortality, increased costs, and adverse outcomes. However, many clinical studies have shown that neither pocket irrigation nor intravenous antibiotics can effectively prevent CIED infection. Moreover, the wide use of antibiotics to prevent CIED infection has led to escalating antibiotic-resistant associated with various pathogens.
METHODS Therefore, we first obtained cross-linked HA (CHA) with BDDE crosslinking agent, in which the degradation time in vivo could cover the acute infection period of 7–10 days after implantation. Then, we designed an injectable hydrogel via electrostatic interaction between carboxyl groups of CHA and amino groups of CHX, which can control the release of CHX.
RESULTS In vitro study with CHA/CHX hydrogel demonstrated CHX release effectively against (99.999% reduction) S. aureus and E. coli. Furthermore, pocket anti-infection was evaluated in vivo using a rabbit model and examined 1 week later. The results showed that CHA/CHX hydrogel group had no purulence and the S. aureus colonies were much lower than those in the bacterial only group (P<0.01). We have also shown that CHX/CHA hydrogel is relatively non-hemolytic and has excellent biocompatibility in vivo.
CONCLUSIONS In summary, CHA/CHX hydrogel may be excellent candidates as anti-infection materials for CIED pocket infection treatment.
GW31-e1115
Qian Xue1, Ruiyu Wang1, Liang Wang1, Bo Xiong1, Lingjiao Li1, Jun Qian1, Lan Hao2, Zhigang Wang2, Dichuan Liu1, Changming Deng1, Shunkang Rong1, Yuanqing Yao1, Yonghong Jiang1, Que Zhu1, Jing Huang1,3
1Department of Cardiology, The Second Affiliated Hospital of Chongqing Medical University
2Chongqing Key Laboratory of Ultrasound Molecular Imaging, Chongqing Medical University
3Institute of Ultrasound Imaging, Department of Ultrasound, The Second Affiliated Hospital of Chongqing Medical University
OBJECTIVES The purinergic P2X3 receptor in the carotid body (CB) is considered a new target for treating hypertension, although approaches for targeted regulating P2X3 receptor expression are lacking. Here, we explored the feasibility of targeted P2X3 receptor down-regulation in CBs by localized low-intensity focused ultrasound (LIFU)-mediated gene delivery to reduce the blood pressure.
METHODS Thirty-two Kunming canines were randomly assigned to the treatment group (n=14), negative control group (n=10), LIFU+cationic microbubbles group (n=4), and LIFU-only group (n=4). Plasmid-loaded cationic microbubbles were injected and bilateral CBs were irradiated with a LIFU-based transducer.
RESULTS Flow cytometry showed that 33.15% of transfected cells expressed the green fluorescent protein reporter gene. T7 endonuclease I assays showed an insertion-deletion rate of 8.30%. The P2X3 receptor mRNA- and protein-expression levels in CBs decreased by 56.31% and 45.10%, respectively, in the treatment group. Mean systolic (152.5±3.0 versus 138.0±2.9 mmHg, P=0.003) and diastolic (97.8±1.5 versus 87.2±2.3 mmHg, P=0.002) blood pressures reduced on day 14 in the treatment group, compared with the baseline values, whereas no effects were observed with LIFU treatment or cationic microbubbles injection alone. Canines treated with this strategy exhibited no local or systemic adverse events.
CONCLUSIONS Thus, LIFU-mediated gene delivery to CBs successfully modulated CB function and reduced blood pressure in a canine model, suggesting a new possibility for treating hypertension and further clinical translation.
GW31-e1118
Wenjun Yan1, Youhu Chen1, Yunlong Xia1, Xinliang Ma2, Ling Tao1
1Department of Cardiology, Xijing Hospital, Fourth Military Medical University
2Department of Emergency Medicine, Thomas Jefferson University
OBJECTIVES Poor engraftment of intramyocardial stem cells limits their therapeutic efficiency against myocardial infarction (MI)-induced cardiac injury. Transglutaminase cross-linked Gelatin (Col-Tgel) is a tailorable collagen-based hydrogel that is becoming an excellent biomaterial scaffold for cellular delivery in vivo. Here, we tested the hypothesis that Col-Tgel increases retention of intramyocardially-injected stem cells, and thereby reduces post-MI cardiac injury.
METHODS Adipose-derived mesenchymal stem cells (ADSCs) were co-cultured with Col-Tgel in a three-dimensional (3D) system in vitro, and Col-Tgel encapsulated ADSCs were observed using scanning electron microscopy and confocal microscopy. Vitality, proliferation, and migration of co-cultured ADSCs were evaluated. In addition, mice were subjected to MI and were intramyocardially injected with ADSCs, Col-Tgel, or a combination thereof. ADSCs engraftment, survival, cardiac function, and fibrosis were assessed.
RESULTS In vitro MTT and Cell Counting Kit-8 assays demonstrated that ADSCs survive and proliferate up to 4 weeks in the Col-Tgel. In addition, MTT and transwell assays showed that ADSCs migrate outside the edge of the Col-Tgel sphere. Furthermore, when compared with ADSCs alone, Col-Tgel-encapsulated ADSCs significantly enhanced the long-term retention and cardioprotective effect of ADSCs against MI-induced cardiac injury.
CONCLUSIONS In the current study, we successfully established a 3D co-culture system using ADSCs and Col-Tgel. The Col-Tgel creates a suitable microenvironment for long-term retention of ADSCs in an ischemic area, and thereby enhances their cardioprotective effects. Taken together, this study may provide an alternative biomaterial for stem cell-based therapy to treat ischemic heart diseases.
GW31-e1152
Jialin Abuzhalihan, Yi Tong Ma
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, P.R. China
OBJECTIVES The aim of this study was to investigate whether DNA methylation of NPC1L1, LIMA1 and Numb genes were associated with CHD.
METHODS Patients with CHD were screened from The First Affiliated Hospital of Xinjiang Medical University between July 2012 and March 2016. DNA methylation levels of the candidate genes NPC1L1, LIMA1 and Numb were measured in peripheral blood leukocytes (PBLs) from 99 patients diagnosed with CHD and 89 control subjects. A total of 55 CPG sites around promoter regions of them were examined. The data were analyzed by IBM SPSS Statistics Version 23.0 (Armonk, NY: IBM Corp.). The measurement data are shown as the Means±SD, and the differences between CHD and control subjects were assessed using an independent-sample t-test. Differences in the enumeration data, such as the frequencies of smoking, drinking, hypertension and diabetes between CHD and control subjects were analyzed using the chi-square test. The methylation levels of LIMA1, NPC1L1, and NUMB did not meet the normality assumption, they were described as median (interquartile range) and compared with Mann–Whitney U test.
RESULTS The average age of the 188 analyzed patients was 59.46±10.61 years, and 127(67.6%) of them were male. There were 60(31.9%) patients with hypertension and 18(9.6%) with DM. Compared with patients without CHD, those with CHD had higher prevalence of HTN (40.4 vs. 22.5%, P=0.008) and T2DM (14.1 vs. 4.5%, P=0.025). Patients with CHD had higher TC (4.52 vs. 4.02 mmol/L, P=0.001), LDL (2.74 vs. 2.59 mmol/L, P=0.013), and TG (1.65 vs. 1.43 mmol/L, P=0.034) levels. The methylated levels of 4 sites in LIMA1, 1 site in NPC1L1, and 4 sites in Numb, respectively, were higher in CHD group than the control group. Compared with control subjects, patients with CHD had higher methylation levels of Numb (P=0.024). After adjustment of male, age, smoking, drinking, hypertension, diabetes, TG, TC, HDL-C and LDL-C, NUMB methylation levels were still associated with CHD (P=0.003, OR=0.260, 95% CI: 0.154–0.568).
CONCLUSIONS Our study showed that Numb methylation may be associated with the development of CHD, and high methylation of Numb gene may be a risk factor for CHD. Patients with higher methylation levels in Numb may have increased risks for CHD. Because our study was an observational study, further studies on the specific mechanisms of this relationship are needed.
GW31-e1154
Dilare Adi, Jialin Abuzhalihan, Yi Tong Ma
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, P.R. China
OBJECTIVES IDOL (inducible degrader of LDLR), also known as MYLIP, is an E3 ubiquitin ligase. It can stimulate the ubiquitination and degradation of the LDLR in the lysosome by interacting with its cytoplasmic domain. In this study, we aimed to explore whether some genetic variants of the IDOL gene were associated with CAD among Chinese population in Xinjiang.
METHODS We designed two independent case-control studies. The first one included in the Han population (448 CAD patients and 343 controls), and the second one is the Uygur population (304 CAD patients and 318 controls). SNP genotyping was performed based on the iMLDR (improved multiplex ligation detection reaction). We genotyped three SNPs (rs2072783, rs2205796, and rs909562) of the IDOL gene. Analyses were carried out using SPSS version 22.0 (SPSS, Chicago, IL). All data were assessed for normality (Kolmogorov-Smirnov test) and equal variance tests. A P value <0.05 was considered to be statistically significant.
RESULTS The genotype distributions of these SNPs met the Hardy-Weinberg equilibrium balance (all P>0.05). Our results showed that, in Han population, the differences in SNP rs2072783 between CAD patients and control subjects were not significant. However, there were significant differences in dominant model (TT vs. GG+GT) and overdominant model (GG+TT vs. GT) of SNP rs2205796 CAD between the two groups (P=0.048 and P=0.029; Table 2). Meanwhile, the dominant model (AA vs. GG+GA) of the rs909562 SNP was significantly different in two groups (P=0.032). Nevertheless, the three SNPs distribution did not show differences in Uygur population. According to the results of the multivariate adjustments for the confounders in Han population, the rs2205796 SNP is an independent risk factor for CAD [TT vs. GG/GT: odds ratio=0.834, 95% confidence interval=0.701–0.993, P=0.042]. Whereas after adjustment for other confounders, rs2072783 and rs909562 SNPs are not the independent risk factors for CAD (P=0.698 and P=0.098). The three SNPs do not represent the independent risk factors for CAD in Uygur population (all P>0.05).
CONCLUSIONS The rs2205796 polymorphism of the IDOL gene is associated with CAD in the Chinese Han population. Subjects with GG/GT genotype or G allele of rs2205796 were related to an increased risk of CAD.
GW31-e1157
Jing Ming, Yi Tong Ma
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, P.R. China
OBJECTIVES Coronary artery disease (CAD) is a multifactorial disease. In this study, we used Xinjiang Han and Uygur populations as the research objects to screen and study the correlation between the genetic polymorphism of the RNF145 gene and CAD.
METHODS We designed two independent case-control studies. The first one included in the Han population (508 CAD patients and 512 controls), and the second one is the Uygur population (410 CAD patients and 330 controls). We used the modified multiple connection detection response (iMLDR) technology to label three SNPs (rs17056583, rs12188266, rs7732603) of RNF145 genotyping.
RESULTS In Uygur group and Han group, for three genotypes of three SNPs, we discovered that there was a difference between CAD and controls in the dominant model, recessive model, and additive model (All P<0.05). The recessive model (GC+GG vs. CC) of rs17056583 and rs7732603 CC genotype remain significantly associated with CAD after adjustment for confounders (OR=1.506, 95% CI=1.319–1.779, P=0.025; OR=1.782, 95% CI=1.636–2.027, P=0.032).
CONCLUSIONS The three SNPs of the RNF145 gene are associated with CAD in the Chinese Han and Uygur population.
GW31-e1194
Feiran Qi1, Yanzhenzi Zhang1, Ke Xu1, Huinan Zhao1, Yan Liu1, Jianxin Chen2, Yulin Li1, Jie Du1
1Beijing Anzhen Hospital of Capital Medical University and Beijing Institute of Heart Lung and Blood Vessel Diseases
2Beijing University of Chinese Medicine
OBJECTIVES Thoracic aortic aneurysms and dissections (TAAD) are highly lethal diseases without effective drug therapeutic strategies. Multiple previous studies in animals and humans indicated a major role of matrix metalloproteinases (MMPs) in TAAD pathogenesis, but clinical trials of MMPs inhibitors for aneurysms all had negative results, which might result from the disadvantages of synthetic compounds. By using an artificial intelligence technology, we discovered that Crocin, a small molecule compound from herbal drug saffron crocus, is a novel MMPs inhibitor. And then we used a mouse model to investigate whether Crocin could attenuate TAAD progression.
METHODS We used surflex-Dock technology to select potential MMP inhibitors from natural products. The effect of crocin on MMPs activity was evaluated by generic MMPs activity kit, gelatin zymography and transwell tumour invasion assay. Biofilm interference technique (BLI) was used to indicate the possible way of interaction between crocin and MMPs. TAAD mouse model was established by 3-aminopropionitrile (BAPN) treatment (1 g/kg d) for 28 days in C57BL/6 mice. Crocin (50 mg/kg d, 80 mg/kg d) was given by intraperitoneal injection from the first day or the 14th day of BAPN treatment to the end of experiments. Aortic structure was evaluated by elastic tissue stain and situ MMPs activity on the aorta wall was detected by the Gelatinase/Collagenase Assay Kit.
RESULTS Crocin was estimated to have a high structural compatibility with MMP1, MMP2, MMP3, MMP10 and MMP14. Then we found crocin inhibited the activity of generic MMPs in Hep G2 cells and remarkably reduced the invasion amount of SL4 tumor cells. The result of BLI showed that crocin had a high affinity for MMP2 on the structure. It also inhibited the activity but not the mRNA or protein expression of MMP2 in HepG2 cells. Furthermore, we observed that either 28 days or 14 days of crocin treatment could suppress TAAD occurrence and rupture. Meanwhile, crocin significantly decreased aortic diameter, ameliorated integrity of vascular structure, inhibited elastin fragmentation and MMPs activity on the aorta wall.
CONCLUSIONS These results indicate that Crocin was a novel broad-spectrum MMPs inhibitor and attenuated TAAD progression in a mouse model, which demonstrated that Crocin was a promising agent for the pharmacological treatment of TAAD.
GW31-e1231
Chen Lin, Yongzhen Guo, Yunlong Xia, Wenjun Yan, Ling Tao
Department of Cardiology, Xijing Hospital, the Fourth Military Medical University, Xi’an, Shaanxi, China
OBJECTIVES Irisin, the cleaved protein of fibronectin type III domain containing 5 (FNDC5), plays a regulatory role in metabolism and inflammation. Recent findings indicate that irisin is involved in cardiovascular physiology and pathology. In the present study, we investigated the effect of FNDC5/irisin on diabetic cardiomyopathy (DCM) in type 2 diabetes mellitus (T2DM) model, db/db mice.
METHODS Myocardial FNDC5 overexpression in 16-week-old db/db mice was achieved by intramyocardial injection of adenovirus encoding FNDC5. Sixteen-week-old db/db mice received recombinant human irisin administration through peritoneal implant osmotic pumps for 4 weeks. Systemic insulin resistance was evaluated by glucose tolerance test. Cardiac diastolic function was evaluated by Doppler echocardiography. Systolic function was assessed by M-mode echocardiography. Left ventricular tissue was collected for histology and gene/protein expression analysis. Cardiac mitochondria and ultrastructure were evaluated by transmission electron microscope. Primary cardiomyocytes or H9C2 cells were treated with high glucose (25 mM)/high fat (palmitate, 300 μM) with or without irisin (1000 ng/mL) for 24 hours. DHE staining and DHR-123 staining were used to detect reactive oxygen species (ROS) and ONOO-, respectively. JC-1 staining was applied to evaluate mitochondrial membrane potential. Expression of 3-nitrotyrosine was measured by western blot to evaluate nitrosative damage in different groups of cells.
RESULTS Compared with db/+ hearts, db/db hearts showed decreased FNDC5 level and exhibited normal cardiac systolic function but impaired diastolic function with adverse structural remodeling, including increased myocardial fibrosis, cardiac hypertrophy and myocardial apoptosis. As compared to db/+ hearts, db/db hearts showed excessive mitochondrial fission and abnormal mitochondrial morphology. Both myocardial FNDC5 overexpression or exogenous irisin administration markedly attenuated cardiac diastolic dysfunction and structural remodeling observed in db/db hearts. The mRNA and protein expression of FNDC5 were significantly decreased in high glucose/high fat treated primary cardiomyocytes and H9C2 cells as compared with control groups. Oxidative/nitrosative stress was evident in high glucose/high fat treated H9C2 cells, as evidenced by elevated iNOS, NOX2, 3-nitrotyrosine, ROS and ONOO- levels. Irisin alleviated oxidative/nitrosative stress induced by high glucose/high fat treatment. Moreover, mitochondria membrane potential (ΔΨm) was decreased and cytochrome C was released from mitochondria with increased caspase-3 activities in high glucose/high fat treated H9C2 cells, indicating the presence of mitochondrial dependent apoptosis. Irisin treatment partly reversed the mitochondrial dependent apoptosis in high glucose/high fat treated H9C2 cells.
CONCLUSIONS Our data demonstrate that FNDC5/irisin exerts a cardioprotective role in diabetic cardiomyopathy in T2DM by alleviating insulin resistance, inhibiting myocardial apoptosis, myocardial fibrosis, and cardiac hypertrophy. FNDC5/irisin restored the diastolic function in T2DM. The attenuation of oxidative/nitrosative stress and apoptosis may be the possible mechanism involved in the process. These findings suggest that FNDC5/irisin may be a potential therapeutic intervention for DCM, especially in type 2 diabetes mellitus.
GW31-e1276
Ruiyu Wang, Jing Huang
Department of Cardiology, The Second Affiliated Hospital of Chongqing Medical University
OBJECTIVES Baroreflex sensitivity (BRS) is impaired in patients with hypertension and is closely related to the increased oxidative stress that occurs in hypertension. As a major redox protein, thioredoxin (TRX) exerts regulatory effects on vascular biology by maintaining redox homeostasis in many cardiovascular diseases. However, the potential association between TRX and BRS under hypertension conditions is currently unknown.
METHODS We evaluated hemodynamic indexes, including pulse arterial pressure (PAP), mean blood pressure (MBP), heart rate (HR), heart rate variability (HRV), and cardiac BRS index in the two-kidney one-clip (2K1C) hypertensive rat model. Several representative biomarkers of oxidative stress and TRX system-related proteins were also evaluated. Additionally, the quantitative expression of TRX in carotid artery homogenates and plasma was detected, and the correlations among TRX, BRS and MBP were identified by regression analyses.
RESULTS BRS was considerably impaired in hypertensive rats, in association with a systemic and vascular oxidative imbalance. In addition, a dysfunctional vascular TRX redox system was observed in hypertensive rats. Additionally, the expression and distribution of TRX protein around the carotid sinus was significantly downregulated in hypertensive rats. The regression analysis showed that the protein level of TRX in the carotid artery homogenates was positively correlated with cardiac BRS index in all rats (|r|=0.69, P<0.001), and negatively correlated with MBP in hypertensive rats (r=-0.43, P<0.05).
CONCLUSIONS TRX is closely related to BRS, and TRX may serve as a potential regulatory protein in attempts to control blood pressure through baroreflex modulation.
CLINICAL RESEARCH ON CARDIOVASCULAR DISEASES
CORONARY HEART DISEASE
GW31-e0020
Ru Liu1, Tianyu Li1, Deshan Yuan1, Yan Chen1, Xiaofang Tang1, Lijian Gao1, Ce Zhang1, Sida Jia1, Pei Zhu1, Ou Xu2, Runlin Gao1, Bo Xu1, Jinqing Yuan1
1Fuwai Hospital, Chinese Academy of Medical Sciences
2Department of Pulmonary Vascular and General Medicine, Fuwai Yunnan Cardiovascular Hospital
OBJECTIVES This study analyzed the association between on-treatment platelet reactivity and long-term outcomes of patients with acute coronary syndrome (ACS) and thrombocytopenia (TP) in the real world.
METHODS A total of 10,724 consecutive cases with coronary artery disease who underwent percutaneous coronary intervention (PCI) were collected from January to December 2013. Cases with ACS and TP under dual anti-platelet therapy were enrolled from the total cohort. Five-year clinical outcomes were evaluated among cases with high on-treatment platelet reactivity (HTPR), low on-treatment platelet reactivity (LTPR) and normal on-treatment platelet reactivity (NTPR), tested by thromboelastogram (TEG).
RESULTS Cases with HTPR, LTPR and NTPR accounted for 26.2, 34.4 and 39.5%, respectively. Cases with HTPR were presented with the most male sex, lowest hemoglobin level, highest erythrocyte sedimentation rate and most LM or three-vessel disease, compared with the other two groups. The rates of 5-year all-cause death, major adverse cardiovascular and cerebrovascular events (MACCE), cardiac death, myocardial infarction (MI), revascularization, stroke and bleeding were all not significantly different among three groups. Multivariable Cox regression indicated that, compared with cases with NTPR, cases with HTPR were not independently associated with all endpoints, as well as cases with LTPR (all P>0.05).
CONCLUSIONS In patients with ACS and TP undergoing PCI, 5-year all-cause death, MACCE, MI, revascularization, stroke and bleeding risk were all similar between cases with HTPR and cases with NTPR, tested by TEG, in the real world. The comparison result was the same between cases with LTPR and cases with NTPR.
GW31-e0021
Ru Liu1, Yang Hu2, Jingang Yang1, Qingsheng Wang3, Hongmei Yang3, Zhifang Wang4, Shuhong Su4, Jinqing Yuan1, Yuejin Yang1
1Department of Cardiology, Fuwai Hospital, Chinese Academy of Medical Sciences
2Statistics Medical Research and Biometrics Center, Fuwai Hospital, Chinese Academy of Medical Sciences
3Department of Cardiology, The First Hospital of Qinhuangdao City
4Department of Cardiology, The Central Hospital of Xinxiang
OBJECTIVES This study analyzed the association of baseline thrombocytopenia (TP) with long-term outcomes of patients with acute ST-segment elevated myocardial infarction (STEMI).
METHODS A total of 16,957 consecutive cases with STEMI from multiple centers that participated in the China Acute Myocardial Infarction registry was included. Two-year clinical outcomes were evaluated between patients with TP and those with a normal platelet count. Possible confounders in baseline were adjusted in multivariable Cox regression. A propensity score matching (PSM) analysis was applied to control baseline differences.
RESULTS Cases coexisting with baseline TP accounted for 2.1%, and cases with moderate or severe TP accounted for 0.3%. The rates of 2-year all-cause death and major adverse cardiovascular and cerebrovascular events (MACCE) were significantly higher in cases with TP (21.4 and 11.4%, P<0.001; 23.6 and 13.9%, P<0.001), compared with those with a normal platelet count. After multivariate adjustment, compared with cases with a normal platelet count, cases with TP was independently associated with 2-year all-cause death (HR: 1.28; 95% CI: 1.02–1.60; P=0.037). After PSM, the rates of 2-year all-cause death and MACCE were significantly higher in cases with TP (20.5 and 14.0%, P=0.022; 22.8 and 16.0%, P=0.021), compared with those with a normal platelet count. Kaplan-Meier survival curves before and after PSM revealed the consistent results. Multivariable Cox regression after PSM showed baseline TP was an independent predictor of all-cause death (HR: 1.59; 95% CI: 1.11–2.29; P=0.013) and MACCE (HR: 1.51; 95% CI: 1.07–2.13; P=0.019).
CONCLUSIONS Baseline TP was an independent predictor of long-term ischemic outcomes in patients with STEMI.
GW31-e0035
Xiaoqing Cai1,2, Feng Tian1, Jing Jing1, Yundai Chen1
1Department of Cardiology, The First Medical Center of PLA General Hospital, Beijing, China
2Department of Cardiology, The 940th Hospital of Joint Logistics Support Force of PLA, Lanzhou, China
OBJECTIVES The study aimed to scientifically evaluate the risk factors of recurrent restenosis after drug-coated balloon (DCB) angioplasty for drug-eluting stent (DES) restenosis among patients enrolled in the RESTORE ISR China randomized trial.
METHODS Patients undergoing the RESTORE ISR China randomized trial with the follow-up angiography were enrolled and classified into the recurrent restenosis group and the non-recurrent restenosis group. The binary classifications followed the QCA standards of in-stent restenosis (ISR): diameter stenosis ≥50% in the in-segment area at follow-up angiography as recurrent restenosis. Clinical and angiographic characteristics of these two groups were analyzed, and the QFR value both before lesion preparation and after final DCB angioplasty were also measured and compared.
RESULTS Two hundred and twenty-six lesions in 208 patients with a follow-up angiography at 9 months were enrolled. Recurrent restenosis was detected in 43 patients (20.7%) and in 49 lesions (21.7%). QFR value after DCB angioplasty (odds ratio [OR] 0.88; 95% confidence interval [CI] 0.83–0.93; P<0.0001 for 0.01 increase), lesion length (OR: 1.08; 95% CI: 1.01–1.15; P=0.017 for 1 mm increase), and vessel diameter (OR: 0.35; 95% CI: 0.13–0.89; P=0.027 for 1 mm increase) were independently associated with recurrent restenosis after DCB angioplasty.
CONCLUSIONS QFR value after DCB angioplasty, lesion length and vessel caliber were independent risk factors of recurrent restenosis after DCB angioplasty. Furthermore, QFR value after DCB angioplasty was a novel and promising predictor in evaluating prognosis after DCB angioplasty of DES ISR.
GW31-e0037
Yuzhou Xue, Jian Shen, Zhou Wei, Xiang Li, Jing Xiang, Zhenxian Xiang, Yuansong Zhu, Haonan Yang, Suxin Luo
The First Affiliated Hospital of Chongqing Medical University
OBJECTIVES Coronary atherosclerotic burden in patients with ST-segment elevation myocardial infarction (STEMI) is recognized as the main predictor of prognosis. However, the association of lipoprotein (a) [Lp(a)] with atherosclerotic burden is uncertain for patients with STEMI.
METHODS Patients with STEMI were continuously enrolled in our study. Multivariate logistic regression analysis was used to assess the relationship between Lp(a) and atherosclerotic burden after adjusted for traditional cardiovascular risk factors. Generalized additive model and restricted cubic spline analyses were employed to visualize the association of Lp(a) with Gensini score and no-reflow phenomena respectively. Kaplan-Meier curve was conducted to explore whether Lp(a) is a predictor of long-term follow up.
RESULTS One thousand and three hundred and fifty-nine patients underwent PCI for STEMI were ultimately included in our study. Patients in the highest tertile of Lp(a) group had increased incidence of acute kidney injury and heart failure during hospitalization. Furthermore, patients with high levels of Lp(a) (>19.1 mg/dL) had sharply increased risk for higher Gensini score (Pfor trend=0.03) and no-reflow phenomena (Pfor trend=0.002) after adjusted. During a median follow up of 930 (interquartile range: 579–1243) days, 132 deaths (9.95%) were registered. Patients with high values of Lp(a) (>19.1 mg/dL) had the worst long-term prognosis (Pfor trend<0.0001). In the subgroup analysis, the patients with high Lp(a) still had the highest all-cause mortality.
CONCLUSIONS This was a new finding that Lp(a) was independently associated with coronary atherosclerotic lesions and prognosis in STEMI patients treated with PCI.
Clinical Trial Registration: identifier ChiCTR1900028516 (http://www.chictr.org.cn/index.aspx).
GW31-e0038
Yuzhou Xue, Yuansong Zhu, Jian Shen, Wei Zhou, Jing Xiang, Zhenxian Xiang, Linbang Wang, Suxin Luo
The First Affiliated Hospital of Chongqing Medical University
OBJECTIVES Cardiogenic shock (CS) is the leading cause of the death in patients with ST elevation myocardial infarction (STEMI). Thyroid dysfunction is related to prognosis of patients with myocardial infarction. Hence, the aim of this study is to explore the relationship between thyroid hormones (free triiodothyronine [FT3] and free thyroxine [FT4]) and CS.
METHODS One thousand two hundred and seventy STEMI patients treated with percutaneous coronary intervention (PCI) were consecutively enrolled in our study. Patients were classified into two groups depend on with or without CS during hospitalization. Stepwise multivariate logistic analysis was conducted to investigate the association of thyroid hormones with CS. Restricted cubic spline method was employed to further explore relationship between CS and thyroid hormones.
RESULTS Patients who developed CS (n=103) had lower FT3 and higher FT4 on admission. The stepwise logistic analysis showed both FT3 (P=0.038) and FT4 (P=0.024) were independently related to CS. Restricted cubic splines indicated that lower FT3 (<2.25 pg/mL) or higher FT4 (>1.25 ng/dL) was correlated with higher prevalence of CS. Over 2.5 years follow-up, patients (n=294) with low FT3 (<2.85 pg/mL) and high FT4 (>=0.88 ng/dL) had the highest all-cause mortality (18.2%), whereas patients (n=293) with high FT3 and low FT4 had the lowest all-cause mortality (3.8%) (Pfor trend<0.0001).
CONCLUSIONS Both FT3 and FT4 are independently associated with in-hospital CS development in STEMI patients treated with PCI. Patients with lower range of FT3 and upper range of FT4 had the worst outcomes in a long-term follow-up.
Clinical Trial Registration: identifier: ChiCTR1900028516 (http://www.chictr.org.cn).
GW31-e0046
Qi Guo1,2, Maoxiong Wu1,2, Hongwei Li1,2, Huijun Ouyang1,2, Runlu Sun1,2, Junjie Wang1,2, Zhijian He1,2, Yuling Zhang1,2, Jingfeng Wang1,2
1Department of Cardiology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University, Guangzhou, China
2Guangdong Province Key Laboratory of Arrhythmia and Electrophysiology, Guangzhou, China
OBJECTIVES Myocardial infarction (MI) is a leading cause of mortality worldwide, and prognostic prediction is critical for identifying high-risk patients and making decisions regarding treatment. We aimed to develop and validate a prognostic nomogram in order to improve the prediction of survival of critically ill MI patients.
METHODS Critically ill patients with MI were identified from the Medical Information Mart for Intensive Care (MIMIC)-III database. Factors included in the nomogram were determined by univariate and multiple Cox proportional hazard analyses based on the primary cohort. The receiver operating characteristic (ROC) and calibration curves were used to assess the predictive accuracy and discriminative ability of the nomogram. The clinical utility of the nomogram was evaluated using decision curve analysis (DCA) and survival curve analysis in an independent validation cohort.
RESULTS Independent prognostic factors, including age, heart rate, white blood cell count (WBC), blood urea nitrogen (BUN) level, and bicarbonate level, were identified and used in the nomogram. Good agreement between the prediction by the nomogram and the actual observation was indicated by the calibration curve for 30-day survival. In the primary cohort, the area under the ROC curve [AUC, 95% confidence interval (CI)] and the C-index (95% CI) were 0.803 (0.771–0.835) and 0.787 (0.757–0.817), respectively. In the validation cohort, the nomogram still exhibited excellent discrimination [AUC (95% CI), 0.765 (0.716–0.814)] and good calibration [C-index (95% CI), 0.758 (0.712–0.804)]. DCA demonstrated that the nomogram was clinically beneficial. Additionally, participants could be classified into two risk groups (low and high) by the nomogram, and the 30-day survival probability was significantly different between these groups (P<0.001).
CONCLUSIONS This 5-factor nomogram can accurately predict 30-day survival in critically ill MI patients and might be helpful for risk stratification and decision making for MI patients undergoing clinical treatment.
GW31-e0047
Qi Guo1,2, Hongwei Li1,2, Huijun Ouyang1,2, Runlu Sun1,2, Junjie Wang1,2, Maoxiong Wu1,2, Yue Pan1,2, Jingfeng Wang1,2, Yuling Zhang1,2
1Department of Cardiology, Sun Yat-sen Memorial Hospital of Sun Yat-sen University
2Guangdong Province Key Laboratory of Arrhythmia and Electrophysiology, Guangzhou, China
OBJECTIVES Whether heart rate (HR) fluctuation after admission has an impact on the outcomes of critically ill myocardial infarction (MI) patients remains unknown.
METHODS A total of 2031 MI patients were enrolled from the Medical Information Mart for Intensive Care (MIMIC-III) database. HR fluctuation was calculated as the maximum HR minus the minimum HR in the initial 24 hours after admission. Participants were divided into 3 groups, namely, low HR fluctuation (<30 beats per minute (bpm)), medium HR fluctuation (30–49 bpm), and high HR fluctuation (≥50 bpm). The main outcomes were 30-day and 1-year mortality. Cox regression and restricted cubic spline model were used.
RESULTS Each 10-bpm increase in HR fluctuation was associated with a higher risk of 30-day mortality and 1-year mortality, with hazard ratios of 1.214 (95% CI, 1.179–1.250) and 1.193 (95% CI, 1.164–1.222), respectively. Compared with the low HR fluctuation group, the high HR fluctuation group suffered a significantly higher risk of mortality after adjustment, with hazard ratios of 1.858 (95% CI, 1.277–2.701) for 30-day mortality and 1.620 (95% CI, 1.221–2.151) for 1-year mortality. A typical J-type curve was observed in restricted cubic splines for the association between HR fluctuation and 30-day or 1-year mortality of MI patients, with the lowest risk on the HR fluctuation of 30 bpm. Sensitivity analyses emphasized the robustness of our results.
CONCLUSIONS This retrospective cohort study revealed an independent positive association between HR fluctuation and 30-day and 1-year mortality in critically ill MI patients, which warrants further investigation.
GW31-e0051
Litao Wang1,2, Yining Dai2, Jiyan Chen2, Ling Xue2, Pengcheng He1,2, Yuanhui Liu2, Ning Tan1,2
1Guangdong Provincial People’s Hospital, School of Medicine, South China University of Technology, Guangzhou, China
2Department of Cardiology, Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Coronary Heart Disease Prevention, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China
OBJECTIVES Thrombocytopenia has been reported to be associated with several adverse events, however, there is little data on the prognostic value of thrombocytopenia for infection in patients with acute myocardial infarction (AMI) undergoing percutaneous coronary intervention (PCI). We evaluated the association between thrombocytopenia and infection in such patients.
METHODS A total of 1401 consecutively admitted AMI patients treated with PCI were enrolled between January 2010 and June 2016, with a median follow-up of 2.85 years. All patients were divided into two groups according to the presence (n=186) or absence (n=1215) of thrombocytopenia based on the platelet counts at admission. The primary endpoint was the development of infection during hospitalization, and the major adverse clinical events (MACE) and all-cause death were considered as secondary endpoints.
RESULTS During hospitalization, 186 (13.3%) patients with AMI following PCI were diagnosed with thrombocytopenia (platelet count <150×109/L) in our study. The prevalence of in-hospital infection was significantly higher in the thrombocytopenic group (30.6 versus 16.2%, P<0.001) compared to non-thrombocytopenia group. Similarly, the incidences of in-hospital MACE (30.1 versus 16.4%, P<0.001) and in-hospital death (8.1 versus 3.8%, P=0.008) revealed an increased trend among thrombocytopenia patients. The multivariate analysis indicated that thrombocytopenia was independently associated with in-hospital infection (odds ratio (OR), 2.09; 95% confidence interval (CI), 1.32–3.27; P=0.001). Similar consequences were detected in MACE (OR, 1.92; 95% CI, 1.27–2.87; P=0.002) but not all cause death (OR, 1.87; 95% CI, 0.88–3.78; P=0.091). After follow up with a median time of 2.85 years, patients with thrombocytopenia were not associated with all-cause death (adjusted hazard ratio (HR)=1.19, 95% CI, 0.80–1.77, P=0.383).
CONCLUSIONS Thrombocytopenia, as a common symptom in patients with AMI undergoing PCI, is significantly correlated with infection and in-hospital MACE, and may be used as a prognostic tool in these patients.
GW31-e0053
Lei Guo, Lei Zhong, Jian Wu, Haichen Lv, Huaiyu Ding, Jiaying Xu, Rongchong Huang
First Affiliated Hospital of Dalian Medical University
OBJECTIVES Limited data exist regarding the clinical outcomes of the three potential therapeutic strategies of percutaneous coronary intervention (PCI), coronary artery bypass grafting (CABG), and medical therapy (MT). The aim of this study was to compare the clinical outcomes of coronary chronic total occlusion (CTO) patients according to the initial treatment strategy.
METHODS Consecutive patients with at least one coronary CTO were included and categorized as managed either by PCI, CABG, or MT. Propensity-score matching was also performed to adjust for baseline characteristics. The primary outcome was a major adverse cardiac event (MACE), including cardiac death, myocardial infarction, and target vessel revascularization.
RESULTS A total of 1655 patients with 1944 CTOs were enrolled in this study. A CTO was treated by MT in 800 (48.3%) patients and PCI in 734 (44.4%), while 121 (7.3%) underwent CABG. The median overall follow-up duration was 3.6 years. Patients referred for MT had a higher incidence of MACE (28.0 vs. 17.3 and 12.4%, respectively; all P<0.001) than those managed by PCI and CABG. After propensity-score matching analysis, the rate of PCI was lower than that of MACE (19.2 vs. 28.9%, HR=0.62, 95% CI=0.45–0.85, P=0.003) when compared with MT. 286 matched pairs of patients were created for patients undergoing successful PCI or MT. There was no significant difference in the prevalence of MACE (HR 0.76, 95% CI=0.53–1.09, P=0.130) or cardiac death (HR 0.51, 95% CI=0.23–1.15, P=0.104) between the successful PCI and MT groups.
CONCLUSIONS As an initial management strategy for patients with CTOs, PCI or CABG reduced the risk of MACE, as compared with MT alone on follow-up.
GW31-e0062
Yuhong Peng, Leisheng Ru, Haoliang Li, YuYing Zhao, XiaoYing Guo, Yanzhuo Ma
The 980th Hospital of the PLA Joint Logistics Support Force
OBJECTIVES To validate the use of DAPT score for dual antiplatelet therapy in patients with chronic total occlusion (CTO) after percutaneous coronary intervention.
METHODS Patients with CTO who underwent PCI with stent placement and were treated with DAPT for 12 months or prolonged to 12–58 months. The incidence of major cardiovascular and cerebrovascular events (MACCE) and bleeding were valued.
RESULTS A total of 504 patients with CTO who underwent PCI with stent placement were included in the final analysis, median follow-up 34 (28, 44) months. In patients with a DAPT score ≥2, the incidence of MACCE was significantly lower in prolonged group compared with standard group (5.5 vs. 14%, P=0.040). Kaplan-Meier analysis showed that the prolonged group had lower MACCE-free survival rate compared with the standard group (P=0.046). The incidences of cardiac death and target vessel revascularization were significantly lower than standard treatment group (1.8 vs. 8.6%, P=0.046; 1.8 vs. 8.6%, P=0.046, respectively). In patients with a DAPT score <2 points, there was no significant difference in the incidence of MACCE between the two groups. But bleeding events were significantly lower in the standard treatment group than in the prolonged DAPT group (3.4 vs. 12.8%, P=0.018). Kaplan-Meier analysis showed that there was a lower bleeding survival rate in the standard treatment group according to BARC criteria (type 2, 3 or 5) (P=0.034).
CONCLUSIONS DAPT score can be used for dual antiplatelet therapy in patients with CTO after percutaneous coronary intervention. CTO patients with DAPT ≥2 points after intervention with a higher risk of ischemic events, might benefit from prolonged DAPT, while patients with DAPT <2 points might benefit from standard treatment, with a lower risk of bleeding events.
GW31-e0064
Xinya Dai1,2, Yingying Zheng1,2, Jinying Zhang1,2
1Department of Cardiology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, P.R. China
2Key Laboratory of Cardiac Injury and Repair of Henan Province, Zhengzhou, China
OBJECTIVES Alkaline phosphatase (ALP) and albumin (ALB) have been shown to be associated with coronary artery disease (CAD), and it has been reported that alkaline phosphatase to albumin ratio (AAR) is a novel independent predictor for the prognosis of pancreatic ductal adenocarcinoma. However, to our knowledge, the relationship between AAR and long-term adverse outcomes in CAD patients after undergoing percutaneous coronary intervention (PCI) has not been investigated. Therefore, we aim to access the relation between AAR and long-term adverse outcomes in post-PCI patients with CAD.
METHODS Three thousand three hundred and seventy eight post-PCI patients with CAD were enrolled in the retrospective CORFCHD-ZZ study from January 2013 to December 2017. The median duration of follow-up was 37.59 months. The primary endpoint was long-term mortality including all-cause mortality (ACM) and cardiac mortality (CM). The secondary endpoints were major adverse cardiac events (MACEs) and major adverse cardiac and cerebrovascular events (MACCEs).
RESULTS Kaplan-Meier analyses showed that an increased AAR was positively correlated with incidences of long-term ACM (log-rank, P=0.014), CM (log-rank, P=0.011), MACEs (log-rank, P=0.013) and MACCEs (log-rank, P=0.006). Multivariate Cox regression analyses showed that the elevated AAR was an independent predictor of long-term ACM (adjusted HR=1.488 [1.031–2.149], P=0.034), CM (adjusted HR=1.837 [1.141–2.959], P=0.012), MACEs (adjusted HR=1.257 [1.018–1.551], P=0.033) and MACCEs (adjusted HR=1.237 [1.029–1.486], P=0.024).
CONCLUSIONS An elevated AAR is a novel independent predictor of long-term adverse outcomes in CAD patients following PCI.
GW31-e0076
Yong Wang, Aijie Hou, Bo Luan
The People’s Hospital of Liaoning Province
OBJECTIVES Guiding support plays an important role in guidewire and microcatheter coronary channel (CC) tracking in retrograde percutaneous coronary intervention (PCI) therapy for chronic total occlusion (CTO) patients. However, the feasibility and safety of retrograde active use of a mother-and-child catheter are still unclear.
METHODS A total of 271 consecutive CTO patients undergoing retrograde PCI between January 2015 and January 2020 were prospectively analyzed. Clinical data of two groups were compared to explore their feasibility and safety.
RESULTS A total of 271 CTO patients underwent retrograde interventional therapy; 69.0% (187/271) underwent therapy through the septal branch, 31.0% (84/271) through the epicardial collateral channel, and 47.6% (129/271) underwent active retrograde extra backup with a mother-and-child catheter to facilitate retrograde microcatheter collateral channel tracking. The time of wire CC tracking was shorter in the active retrograde backup (ARB) group than in the non-ARB group (25.4±8.5 vs. 26.4±9.7, P=0.348), but there was no significant difference. The time of retrograde microcatheter tracking (10.2±3.8 vs. 15.5±6.8, P=0.012) and time of retrograde approach (62.8±20.3 vs. 70.4±24.3, P=0.026) in the ARB group were significantly shorter than those in the non-ARB group. The radiation dose (223.6±112.7 vs. 295.2±129.3, P=0.028), fluoroscopy time (50.6±21.3 vs. 62.3±32.1, P=0.030), and contrast volume (301.8±146.7 vs. 352.2±179.5, P=0.032) in the ARB group were significantly lower than those in the non-ARB group. There were no serious life-threatening procedural complications in either group. Complications unrelated to ARB included 2 cases of donor vessel dissection, 1 case of CC perforation, and 2 cases of target vessel perforation. There was no statistically significant difference in major adverse cardiac and cerebrovascular events (MACCE) between the groups during hospitalization (P>0.05).
CONCLUSIONS ARB is feasible, safe, and conducive to guidewire and microcatheter coronary channel tracking in recanalization of coronary CTO. It improves the procedural efficiency and is worthy of further promotion.
GW31-e0077
Yong Wang, Aijie Hou, Bo Luan
The People’s Hospital of Liaoning Province
OBJECTIVES Retrograde microcatheter collateral channel (CC) tracking after successful wiring of septal CC is crucial for retrograde revascularization of coronary chronic total occlusion (CTO). However, the incidence, predictors, and strategies for failure of retrograde microcatheter CC tracking after successful wiring of septal CC remain unclear.
METHODS In total, 298 CTO patients who underwent retrograde septal CC PCI between January 2015 and May 2019 were retrospectively analyzed. Clinical data were compared to investigate predictors for initial microcatheter tracking failure.
RESULTS The initial and final microcatheter tracking success rates were 79.2% (236/298) and 96.6% (288/298), respectively. The procedural success rate was 94.0% (280/298). The RCA to LAD septal ratio (48.4 vs. 33.1%, P=0.037) and CC tortuosity (34.6 vs. 20.8%, P=0.045) were significantly higher in the initial microcatheter CC tracking failure group than in the success group. Multivariate logistic regression analysis revealed that severe collateral tortuosity (OR: 13.241, 95% CI: 3.429–27.057, P=0.038), CC entry angle <90° (OR: 4.921, 95% CI: 1.128–9.997, P=0.002), CC exit angle <90° (OR: 5.037, 95% CI: 2.237–11.182, P=0.004), Finecross MG as initial microcatheter (OR: 1.826, 95% CI: 1.127–3.067, P=0.035), and initial retrograde application of guidezilla (OR: 0.321, 95% CI: 0.267–0.915, P=0.024) were independent predictors of initial microcatheter CC tracking failure in patients with CTO undergoing retrograde septal CC PCI.
CONCLUSIONS The initial microcatheter CC tracking failure was 20.8% in total. Severe collateral tortuosity, CC entry and exit angle <90°, Finecross MG as initial microcatheter, and initial retrograde application of guidezilla were independent predictors of initial microcatheter CC tracking failure in patients with CTO undergoing retrograde septal PCI.
GW31-e0078
Yong Wang, Aijie Hou, Bo Luan
The People’s Hospital of Liaoning Province
OBJECTIVES Patients with coronary chronic total occlusion (CTO) require effective antiplatelet therapy after percutaneous coronary intervention (PCI). Ticagrelor has more pronounced platelet inhibition than clopidogrel. However, the most appropriate dose of ticagrelor in East Asian populations remains unclear.
METHODS We compared ticagrelor (180 mg loading dose, 90 mg twice daily thereafter and 120 mg loading dose, 60 mg twice daily thereafter) and clopidogrel (300 mg loading dose, 75 mg daily thereafter) for prevention of cardiovascular events in 525 patients with CTO undergoing PCI.
RESULTS The rate of in-hospital major adverse cardiac and cerebral events (MACCE) was not different between the groups. At 1-year follow-up, target vessel revascularization (TVR) in both ticagrelor groups were significantly lower than that in the clopidogrel group (P=0.047); TVR was significantly decreased in 60 mg ticagrelor compared to standard dose clopidogrel (P=0.046). At 1-year follow-up, overall MACCE in both ticagrelor groups were significantly lower than that in the clopidogrel group (P=0.023). Kaplan–Meier analysis showed MACCE-free survival was significantly higher in both ticagrelor groups than in the clopidogrel group (P=0.024). During hospitalization, minor bleeding was significant increased in the 90 mg ticagrelor group (P=0.021). At 1-year follow-up, risk of major and minor bleeding were significantly increased in the 90 mg ticagrelor group.
CONCLUSIONS In East Asian patients with CTO undergoing PCI, 60 mg ticagrelor was as effective as 90 mg, at the same time significantly reduced risk of bleeding.
GW31-e0079
Yong Wang, Aijie Hou, Bo Luan
The People’s Hospital of Liaoning Province
OBJECTIVES The incidence and prognosis of coronary slow flow (CSF) and no reflow phenomenon (NRP) in patients with coronary chronic total occlusion (CTO) who underwent percutaneous coronary intervention (PCI) remains unclear.
METHODS We conducted a single-center prospective study to investigate the incidence of CSF/NRP during CTO interventional therapy, determine predictors of CSF/NRP, and evaluate the effect on patient outcomes.
RESULTS In this study, 552 patients with CTO who underwent PCI were included. CSF/NRP occurred in 16.1% of the patients. They had higher incidence rates of diabetes mellitus (53.9 vs. 36.3%, P=0.002) and hypertension (50.6 vs. 37.1%, P=0.018), and a lower incidence rate of retrograde filling of grade >2 (34.8 vs. 47.1%, P=0.036). Patients with CSF/NRP had a higher neutrophil ratio (55.6±19.4 vs. 52.4±18.3, P=0.038) and levels of low-density lipoprotein (LDL; 3.0±0.8 vs. 2.8±0.6, P=0.029), fasting glucose (8.3±1.3 vs. 6.8±1.1, P=0.005), uric acid (332.6±82.9 vs. 308.2±62.8, P=0.045), and high-sensitivity C-reactive protein (Hs-CRP; 9.8±4.8 vs. 7.3±3.9, P=0.036). A multivariate logistic regression analysis revealed that diabetes mellitus (odds ratio [OR], 95% confidence interval [CI]: 1.962, 1.198–2.721, P=0.042), mean platelet volume (MPV; OR: 1.284; 95% CI: 1.108–1.895, P=0.046), LDL cholesterol (LDL-C; OR: 1.383, 95% CI: 1.105–2.491, P=0.036), fasting glucose (FG; OR: 2.095, 95% CI: 1.495–2.899, P=0.018), Hs-CRP (OR: 2.218, 95% CI: 1.556–3.519, P=0.029), and retrograde filling of grade >2 (OR: 0.822, 95% CI: 0.622–0.907, P=0.037) were independent predictors of CSF/NRP in the CTO patients who underwent PCI. Kaplan-Meier analysis revealed that the patients in the CSF/NRP group had a significantly lower cumulative major cardiac and cerebrovascular events (MACCEs)-free survival than those in the non-CSF/NRP group (P<0.0001).
CONCLUSIONS Of the patients with CTO who underwent PCI, 16.1% developed CSF/NRP and they had a significantly lower cumulative MACCE-free survival rate. Diabetes mellitus; higher levels of MPV, LDL-C, FG, and Hs-CRP; and lower incidence of retrograde filling grade >2 were independent predictors of CSF/NRP in CTO patients who underwent PCI and can therefore be used for risk stratification.
GW31-e0097
Siyuan Li1,2, Yifang Yuan1,2, Ping Zhang2
1Tsinghua University, School of Clinical Medicine
2Beijing Tsinghua Changgung Hospital
OBJECTIVES To compare the differences in cardiopulmonary exercise test (CPET) key variables in healthy participants and patients with increasing atherosclerotic burden, especially non-obstructive coronary artery disease (NOCAD).
METHODS Six-hundred and fourteen symptomatic patients (mean age 60 years old, 42% female) and one-hundred and one healthy volunteers (mean age 37 years old, 58% female) were included and all conducted CPET. Symptomatic patients were divided into NOCAD, obstructive coronary artery disease (OCAD) and acute myocardial infarction (AMI) according to the degree of coronary stenosis on coronary angiography. We compared the differences in CPET key variables among NOCAD, OCAD, AMI and healthy participants.
RESULTS Significant associations were observed between increasing atherosclerotic burden and unfavorable CPET variables, including those related to ischemia (O2 pulse trajectory), cardiac function (CO, HR, HR/WR, HR exercise/rest ratio), prognosis (VO2, predicted% VO2 peak, OUES, VE/VCO2 slope). NOCAD patients had significantly lower VO2 AT (β for NOCAD where the healthy as reference, -1.35; 95% CI -2.16 to -0.54), VO2 peak (β for NOCAD where the healthy as reference, -2.05; 95% CI -3.18 to -0.93), OUES (β for NOCAD where the healthy as reference, -0.24; 95% CI -0.36 to -0.11) in pairwise comparison after adjustment for confounders. NOCAD patients might be associated with higher odds for flattening/downward O2 pulse trajectory pattern (OR, 3.62; 95% CI, 1.08, 12.17). The association between increasing atherosclerotic burden and unfavorable CPET variables might be stronger in male NOCAD.
CONCLUSIONS Unfavorable CPET variables were associated with increasing atherosclerotic burden. There were associations between NOCAD patients and myocardial ischemia as well as malignant prognosis in CPET. CPET can be a potential tool for early identification and assessment of NOCAD.
GW31-e0102
Jieyun You, Liming Gao, Yunli Shen, Xingxu Wang, Qing Wan, Wei Guo, Qi Zhang
Department of Cardiovascular Medicine, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China
OBJECTIVES Left ventricular aneurysm (LVA), as a common complication of acute myocardial infarction (AMI), relates to worse prognosis in patients with AMI. Although primary percutaneous coronary intervention (PCI) has been adopted as the first-line treatment for AMI patients in contemporary era, the predictor and prognosis of LVA in AMI patients treated with primary PCI were still limited, especially in those involved with anterior wall infarction. We aimed to investigate the predictors and prognosis of LVA in patients with acute anterior myocardial infarction treated with primary PCI in contemporary era.
METHODS Altogether 942 consecutive patients with acute anterior myocardial infarction undergoing primary PCI were prospectively enrolled, among which 15.92% were in the LVA group and the rest were in the non-LVA group. The primary endpoint of major adverse cardio-cerebral events (MACCEs) was defined as a composite of cardiac death, cardiogenic shock, target vessel revascularization, and ischemic stroke. Baseline characteristics and 1-year clinical outcomes were compared by Chi-square test, t-test, or Kaplan-Meier survival analysis as appropriate. Multiple logistic regression was applied to predict LVA formation. Receiver operating characteristic (ROC) curves were plotted for the accuracy of the multivariate analysis model.
RESULTS At 1-year clinical follow-up, the primary endpoint of MACCEs was significantly increased in the LVA group than in the non-LVA group (23.33 vs. 7.45%, P<0.01), which was mainly driven by higher incidence of cardiac death (8.00 vs. 2.78%, P<0.01), cardiogenic shock (16.00 vs. 3.03%, P<0.01), target vessel revascularization (5.33 vs. 2.27%, P=0.04), and ischemic stroke (4.00 vs. 1.39%, P=0.03). Longer symptom-to-balloon time [odds ratio (OR): 1.15, 95% confidence interval (CI): 1.10–1.21, P<0.01], increased baseline SYNTAX Score (OR: 1.20, 95% CI: 1.16–1.25, P<0.01) and residual SYNTAX Score (OR: 1.54, 95% CI: 1.37–1.74, P<0.01), impaired left ventricular ejection fraction (OR: 0.89, 95% CI: 0.86–0.93, P<0.01), and persistent ST segment elevation (OR: 1.92, 95% CI: 1.03–3.56, P=0.04) were independent predictors of LVA formation in patients with acute anterior myocardial infarction undergoing primary PCI by multivariate analysis.
CONCLUSIONS In contemporary era, patients with LVA still had worse clinical outcomes. Promote reperfusion response with reduction of symptom-to-balloon time, and achieve complete revascularization may help to prevent LVA and improve the prognosis of patients with acute anterior myocardial infarction treated by primary PCI.
GW31-e0123
Wei Wei, Wang Xingxu, Huang Zhenghao, Luo Yu
Department of Cardiovascular Medicine, East Hospital, Tongji University School of Medicine, Shanghai, China
OBJECTIVES Coronary artery aneurysm (CAA) and coronary artery ectasia (CAE) may be two different types of coronary artery dilatation with unknown etiology. This study aimed to compare the differences between CAA and CAE and to investigate their pathogenesis and the necessity of antiplatelet therapy.
METHODS It was a case-control study. One hundred patients each with confirmed CAA, CAE, and normal coronary artery (NCA) from September 2017 to July 2019 were included. All patients completed examinations of the ankle-brachial index (ABI), pulse wave rate, and carotid ultrasonography; were tested for routine blood, lipid, and immune parameters; and given a routine oral loading dose of antiplatelet drugs before coronary angiography. Blood samples were collected 1 week after the withdrawal of antiplatelet drugs, and vascular inflammatory indexes, platelet activation indexes, thromboelastography, and the platelet aggregation rate were measured. Analysis of variance and the chi-square or Fisher exact test was used in statistical analysis.
RESULTS The mean age and prevalence rate of hypertension were significantly higher in CAA and CAE than in NCA (P<0.05). The perinuclear anti-neutrophil cytoplasmic antibody (ANCA), endothelial-1, matrix metallopeptidase-9, and tumor necrosis factor-α were significantly higher in CAE than in NCA, while cytoplasmic ANCA was appreciably higher in CAE than in CAA (P<0.05). Myeloperoxidase and growth/differentiation factor-15 were significantly higher in CAE than in CAA and NCA (P<0.05). ABI was significantly lower in CAA and CAE than in NCA (P<0.05), low-density lipoprotein/high-density lipoprotein was significantly higher in CAA than in NCA (P<0.05), and the detection rate of carotid artery thickening was significantly higher in CAA than in CAE and NCA (P<0.05). The Gensini and SYNTAX scores were significantly higher in CAA than in CAE (P<0.05). The percentages of CD62P and PAC-1 were significantly higher in CAA and CAE than in NCA (P<0.05). The arachidonic acid aggregation rate in CAA and adenosine 5′-diphosphate aggregation rate in CAE were significantly higher than in NCA (P<0.05). The values of K and R were significantly lower in CAE than in NCA (P<0.05), and α angle was significantly higher in CAE than in NCA. There were no significant differences in the values of K, α angle, and R between CAA and NCA (P<0.05).
CONCLUSIONS CAE was closely related to inflammation, whereas CAA was closely related to atherosclerosis. Platelet activation was present in both diseases; therefore, antiplatelet therapy is recommended.
GW31-e0125
Shuai Zhao2, Qin Wang1, Chenxiang Li2, Kun Lian2
1Department of Pharmacogenomics, Fourth Military Medical University
2Department of Cardiology, Xijing Hospital, Fourth Military Medical University
OBJECTIVES Branched-chain amino acids (BCAA) is an important nutrient which can preserve heart function. Previous study reported that BCAA catabolic defect can promote the development of heart failure. However, the role of plasma BCAA in patients with heart failure with reduced ejection fraction (HFrEF) remains largely unknown. The aim of our study is to evaluate the relationship between plasma BCAA level with Chinese HFrEF patients and whether plasma BCAA level is associated with long-term all-cause mortality.
METHODS We conducted a prospective study on a cohort of HFrEF patients (n=168) who was admitted to our hospital and assessed survival at 5 years. Another 107 healthy people who underwent routine physiological examination were selected as control subjects. Plasma BCAA level and other biochemical and clinical parameters were assayed in the studied population. The end-points were adverse cardiac events, including all-cause mortality and re-hospitalization.
RESULTS Plasma BCAA level was significantly increased as well as total adiponectin (total APN) and higher molecular weight adiponectin (HMW APN) (P<0.01) in HFrEF patients. HFrEF patients with myocardial infarction (MI) indicated a significant elevation of plasma BCAA level when compared with HFrEF but not accompanied with MI patients (r=0.164, P=0.033). In all-population, plasma BCAA level correlated with Total APN (r=0.187, P<0.01), HMW APN (r=0.129, P<0.05), left ventricular ejection fractions (LVEF) (r=0.272, P<0.01), N-terminal pro brain natriuretic peptide (NT-proBNP) (r=0.269, P<0.01) and blood uric acid (BUN) (r=0.269, P<0.01). After a follow-up of 5 years, 125 (74.4%) adverse cardiac events occurred, including 84 (50.0%) all-cause mortality and 41 (24.4%) re-hospitalization. Additionally, we observed that plasma BCAA level was not associated with all-cause mortality or adverse cardiac events in HFrEF patients.
CONCLUSIONS Our study finds that plasma BCAA level is increased in HFrEF patients. However, plasma BCAA level is not an independent predictor of long-term all-cause mortality or adverse cardiac events in HFrEF patients.
GW31-e0133
Kun Lian, Chenxiang Li
Department of Cardiology, Xijing Hospital, The Forth Military Medical University
OBJECTIVES To evaluate the safety and efficiency of extracorporeal membrane oxygenation (ECMO) combined with intra-aortic balloon counter pulsation (IABP) assisted selective percutaneous coronary intervention (PCI) in extremely complex and high-risk coronary heart diseases.
METHODS Fourty three consecutive patients who underwent ECMO combined with IABP assisted selective PCI were reviewed from May 2018 to February 2020. Detailed baseline clinical, angiography, revascularization and follow-up data were collected.
RESULTS A total of 43 patients including 36 (83.7%) male, with the mean age of (66.89±11.47) years and ejection fraction (EF) of (40.23±12.10)%. Previous myocardial infarction was 19 cases (44.19%), previous PCI was 10 cases (23.26%), previous CABG (Coronary Artery Bypass Grafting) was 2 cases (4.65%). There were 26 patients with left main severe lesions (60.47%), 32 patients with chronic total occlusion lesions (74.42%), 16 patients with severe 3-vessel lesions (37.21%). Coronary perforation and death occurred in 1 cases (2.33%). Overall procedural success was 97.67% and 4 cases performed complete revascularization (9.30%). The average procedure time was (346.8±77.03) min, ECMO assisted time was (7.60±3.91) h and IABP assisted time was (71.2±86.9) h. Additionally, postoperative complications was 5 cases (11.63%) and in-hospital major adverse cardiovascular events (MACE) occurred in 2 cases (4.65%). During a median follow-up of (6.55±5.22) months, 30 d MACE rate was 16.28%; EF was markedly improved (40.50 vs. 48.94%, P=0.031) and NYHA was decreased significantly (P=0.008), 2–13 m MACE-free was 100%.
CONCLUSIONS In highly selected patients ineligible for CABG, ECMO and IABP supported selective PCI can be performed with a promising clinical outcome.
GW31-e0137
Kun Lian1, Wei Wang2, Qin Wang2, Shuai Zhao1, Genrui Chen1, Chengxiang Li1
1Department of Cardiology, Xijing Hospital, Fourth Military Medical University
2Department of Pharmacogenomics, Fourth Military Medical University
OBJECTIVES This study set out to assess the long-term outcome of chronic total occlusion treated with percutaneous coronary intervention (CTO-PCI) in Chinese patients with low left ventricular ejection fraction (LVEF) (<40%).
METHODS Consecutive patients undergoing elective PCI of CTO were included at Xijing Hospital from Jan 2012 to Feb 2018. Patients with acute ST-segment elevation myocardial infarction were excluded. Patients were subdivided into 3 groups: group A (LVEF≥50%), group B (LVEF 40–50%), and group C (LVEF<40%). Detailed baseline clinical, angiography, revascularization and long-term follow-up data were collected. Accordingly, data were analyzed for long-term outcome and predictors of successful CTO-PCI in patients with low LVEF.
RESULTS A total of 1197 patients (mean 61.92±10.71 years of age, 83.88% men) underwent CTO PCI attempts. Baseline group C was present in 247 (20.63%) patients. The angiographic success was high (overall 91.9%), but lower in group C (93.84 vs. 94.03 vs. 84.21%, respectively; all P<0.0125). Mean clinical follow-up of 42.25±28.96 months duration was available in 1029 (85.96%) patients including those with group C. The major cardiac events (MACE) was higher in group C when compared with group A (16.41 vs. 30.83%; P<0.001), and all-cause mortality was also higher in group C (11.04 vs. 28.57%; P<0.001). In patients with LVEF<40%, cardiac function improved significantly (P<0.001) after a successful PCI. Further, the independent risk factors of MACE in successful revascularization patients with LVEF <40 patients were history of diabetes (OR=3.813, 95% CI 1.212–11.993, P=0.022) and using long stent (OR=13.633, 95% CI 2.597–71.565, P=0.002).
CONCLUSIONS PCI could represent an effective revascularization strategy achieving good long-term outcome in CTO patients with low LVEF.
GW31-e0143
Wenjie Zuo, Genshan Ma
Department of Cardiology, Zhongda Hospital, School of Medicine, Southeast University
OBJECTIVES Although the J-CTO score is widely used in grading the complexity of chronic total occlusions (CTOs), there remains uncertainty regarding its comparative performance with other predictive scores.
METHODS We searched PubMed, EMBASE, and Cochrane Library systematically for relevant studies reporting diagnostic accuracy of J-CTO from the year 2011 onwards. Estimates of discrimination and calibration would be pooled with a random-effects model if possible. The methodological quality of included studies was assessed using the PROBAST system. When analyzing the relative performance of models, we examined the potential for optimism bias in studies favoring new models and models developed by the same authors.
RESULTS Twenty-one eligible studies (31,410 lesions) and 11 scoring systems were included. The J-CTO score exhibited a strong discriminatory capacity for 30-min wire crossing (pooled C-statistic: 0.76, 95% CI 0.68–0.84) while performing moderately for technical success (0.68, 95% CI 0.61–0.74). Forty-three pairwise comparisons were based on discrimination but only 18 of them (42%) reported a statistical result. Of 23 independent comparisons, the J-CTO score performed better (relative difference >5%) in 10 cases and worse in another 7. However, calibration was inappropriately assessed in most studies, leading to potential risk of bias.
CONCLUSIONS The J-CTO score have useful discrimination in both time-efficient wire crossing and final angiographic success but its validation on calibration needs to be strengthened. To date, there is insufficient evidence supporting the superiority of newly introduced models over the J-CTO score.
GW31-e0166
Baxrom Alyavi, Jamol Uzokov, Akbar Abdullaev
Republican Specialized Scientific and Practical Medical Center of Therapy and Medical Rehabilitation
OBJECTIVES To determine the frequency of development of clopidogrel resistance in patients with coronary heart disease and to identify risk factors for the development of resistance to clopidogrel in patients with coronary heart disease.
METHODS One hundred and twenty patients were enrolled in this study with stable forms of ischemic heart disease who received a standard dose of clopidogrel 75 mg/day for at least a year after PCI. Patients were divided into 3 groups according to the prescription of clopidogrel. Group 1 consisted of patients who did not receive clopidogrel, the second group consisted of patients taking clopidogrel up to 1 year, and the third group consisted of patients taking clopidogrel more than 1 year. An addition, according to the results of determining clopidogrel resistance, patients were divided into 2 groups. Platelet aggregation was measured using a laser analyzer: adenosine diphosphate (ADP) with 1.0 and 5.0 μmol was used. Resistance criterion: ADP ≥72% at 5.0 μmol.
RESULTS 19.0% of patients who took clopidogrel in a standard dose of 75 mg/day were resistant to clopidogrel, and a third of them took clopidogrel for more than a year. According to the results of a study of platelet aggregation activity, the average degree of platelet aggregation with 5.0 μmol of ADP was 81.0% in patients with no reaction to clopidogrel. An inadequate response to clopidogrel was observed more in women and the elderly (55.0 and 68.0%). When analyzing the results of routine laboratory studies in patients with clopidogrel resistance, there was a tendency to higher levels of cholesterol and glucose (P<0.05). Possible risk factors for the development of clopidogrel resistance are hypercholesterolemia and hyperglycemia, since clopidogrel resistance was more common in patients with diabetes (12 out of 18) and obesity (10 out of 19).
CONCLUSIONS Clopidogrel resistant is high in patients with coronary artery disease after PCI, especially on those who suffer from hypercholesterolemia and hyperglycemia with type 2 diabetes mellitus and obesity.
GW31-e0173
Masimova Aysel1, Mekhman Mamedov2
1Central Clinical Hospital of the Armed Forces of Azerbaijan, Baku, Azerbaijan
2National Medical Research Center for Therapy and Preventive Medicine of the Ministry of Health, Moscow, Russia
OBJECTIVES To study the behavioral and biological risk factors for CHD among male military personnel.
METHODS The study included 100 men with CHD. Patients were divided into 2 groups according to their social status. Of these, 60 patients were civilians (group I), and the remaining 56 patients (group II) were military personnel. All patients were treated in the cardiovascular department of the Main Hospital of the Armed Forces (Baku, Azerbaijan). The age range was 30–82 years (average age 56±4 years). The following risk factors were studied: smoking, obesity, arterial hypertension, hypercholesterolemia, stress and anxiety/depression.
RESULTS In the study group, the most common risk factor was hypercholesterolemia – 80% (n=93): 46 were military personnel and 47 civilians. AH was detected in 78% of military personnel and 68% among civilians. Smoking was reliably often detected among military personnel – 68% compared with the control group – 50%. Abdominal obesity and diabetes mellitus were statistically significantly detected among civilians with coronary artery disease (57 and 36% versus 38 and 20%). Psychosomatic disorders (chronic stress, anxiety and depression) were 50% or more pronounced among military personnel compared with civilians.
CONCLUSIONS Thus, in both groups of men with CHD, the most common risk factors are hypertension and hypercholesterolemia. Moreover, between the groups there are some differences in a number of risk factors. Among the military, smoking, chronic stress, anxiety/depression predominated, while among civilians, diabetes and abdominal obesity were more often detected. The data obtained can be used to determine the treatment tactics and secondary prevention of CVD among military personnel.
GW31-e0185
Lingyun Gu1,2, Ruolong Zheng1, Wenlong Jiang1
1Department of Cardiology, Jiangyin People’s Hospital
2Department of Cardiology, Zhongda Hospital, Medical School of Southeast University
OBJECTIVES Patients with acute myocardial infarction (AMI) often result in adverse ventricular remodeling and cardiac arrhythmia. The innate immune system is an important mechanism involved in this process. In this study, we tried to investigate the predictive value of high-mobility group box 1 (HMGB1), a protein involved in the innate immune system, on cardiac dysfunction and new-onset atrial fibrillation in patients with AMI.
METHODS Three hundred and forty eight AMI patients underwent primary percutaneous coronary intervention and 312 control subjects without apparent coronary artery disease were enrolled between January 2016 and December 2018. Serum HMGB-1 level was determined by Enzyme-linked immunosorbent assay (ELISA). Echocardiography, N-terminal prohormone of brain natriuretic peptide and continuous electrocardiographic monitoring were recorded for evaluation of in-hospital cardiac function and new-onset atrial fibrillation (NOAF).
RESULTS During hospitalization, elevated serum HMGB1 expression was associated with increased myocardial necrosis and reduced cardiac function in AMI patients. AMI patients with NOAF presented higher serum HMGB1 levels than AMI patients without NOAF. Patients with higher serum levels of HMGB1 were at increased risk of suffering from NOAF.
CONCLUSIONS In our study, we demonstrated that serum level of HMGB1 is positively related to the levels of LDL, cardiac troponin I, NT-proBNP, LVEF and time of symptom onset to balloon dilatation in AMI patients. We also found that an increased prevalence of post-MI NOAF was observed in patients with higher serum HMGB1 levels, serum HMGB1 level could predict post-MI NOAF. Higher levels of serum HMGB1 may be a new predictive factor for in-hospital cardiac dysfunction and NOAF in AMI patients.
GW31-e0189
Sulan Huang, Guo Ning, Ge Liangqing
The First People’s Hospital of Changde
OBJECTIVES To investigate the risk factors of nosocomial complications in young patients with acute myocardial infarction.
METHODS In this study, 122 young patients with acute myocardial infarction from January 2018 to December 2019 were included according to the study’s inclusion criteria. There were 30 cases with complications and 92 cases without complications. The medical history, test index and coronary angiography were compared between the two groups. Multivariate Logistic regression was used to analyze the risk factors of nosocomial complications in young patients with acute myocardial infarction, and the regression equation of nosocomial complications was established. ROC curve was drawn to evaluate the efficacy of regression equation in evaluating the nosocomial complications in young patients with acute myocardial infarction.
RESULTS Young patients with complications of acute myocardial infarction compared with patients without complications, between the two groups of history of high blood pressure, creatinine, fibrinogen, b-type brain natriuretic peptide precursor, heart ejection fraction and coronary artery lesion number type and scope of coronary lesions and acute myocardial infarction, the difference was statistically significant (P<0.05). Logistic regression analysis showed that creatinine (OR=1.01, 95% CI: 1.001–1.019, P=0.036), number of coronary artery lesions (OR=2.519, 95% CI: 1.185–5.357, P=0.016) and fibrinogen (OR=14.984, 95% CI: 3.863–58.113, P<0.001) were risk factors for nosocomial complications in young patients with AMI. ROC curve analysis results showed that the maximum value of Youden index, AUC, sensitivity and specificity of the regression equation for the assessment of nosocomial complications in young patients with AMI were 0.644, 0.873 (95% CI: 794–0.952, P<0.001), 0.769 and 0.875 respectively.
CONCLUSIONS Creatinine, coronary lesion number and fibrinogen were risk factors for nosocomial complications in young patients with acute myocardial infarction. The established regression equation has a high predictive value for the occurrence of nosocomial complications in young patients with acute myocardial infarction.
GW31-e0190
Bing Huang, Hong Jiang
Department of Cardiology, Renmin Hospital of Wuhan University
OBJECTIVES The outbreak of coronavirus disease 2019 (COVID-19) has rapidly spread worldwide. This study sought to share our experiences with in-hospital management and outcomes of acute myocardial infarction (AMI) before and during the COVID-19 epidemic.
METHODS We retrospectively analyzed consecutive AMI patients, including those with ST-elevation myocardial infarction (STEMI) and non-STEMI (NSTEMI), from February 1, 2020, to April 15, 2020 (case group), and from January 1, 2019 to December 31, 2019 (control group) and conducted a 1:1 ratio-matched case-control study.
RESULTS Fifty-three AMI (31 STEMI, 22 NSTEMI) patients during the COVID-19 epidemic were matched to 53 AMI patients before the epidemic. Baseline characteristics were comparable between the groups. STEMI patients in the case group had a longer delay time, less primary or remedial PCI and more emergency thrombolysis than those in the control group. Less coronary angiography and stenting were performed in AMI patients in the case group than in the control group. Although there were no statistically significant differences in clinical outcomes between the two groups, STEMI patients in the case group had more than a three-fold increase in mortality rates. AMI combined with COVID-19 infection was associated with higher rates of mortality than AMI alone.
CONCLUSIONS The COVID-19 epidemic has resulted in significant reperfusion delays in STEMI patients and has a marked impact on the treatment options selection in AMI patients. This epidemic also results in more than a three-fold increase in mortality rates in STEMI patients, although the differences were not statistically significant.
GW31-e0195
Yuanhui Liu1, Litao Wang2, Ji Yan Chen1, Peng Cheng He1, Chong Yang Duan3, Ning Tan1
1Department of Cardiology, Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Coronary Heart Disease Prevention, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China
2Guangdong Provincial People’s Hospital, School of Medicine, South China University of Technology, Guangzhou, China
3Department of Biostatistics, School of Public Health, Southern Medical University, Guangzhou, China
OBJECTIVES Post-acute myocardial infarction (P-AMI) infection is a serious complication in patients with ST-segment elevation myocardial infarction (STEMI) undergoing percutaneous coronary intervention (PCI). We aimed to develop and validate a new risk score for early prediction of P-AMI infection in such patients.
METHODS A total of 1842 STEMI patients undergoing PCI enrolled between January 2010 and May 2016 served as a derivation cohort, and 1270 STEMI patients enrolled between June 2016 and May 2018 served as external validation cohort. We conducted multivariable logistic regression treating a novel risk score as an independent variable predictive for the primary outcome of interest, P-AMI infection during hospitalization.
RESULTS The incidence of P-AMI infection was 11.8% in development cohort. Seven clinical variables were included to establish the risk score: Age, Killip classification, WBC count, serum albumin, insulin use, diuretic use, and transfemoral approach. The risk score demonstrated high discrimination, with a C-statistic of 0.851 (95% CI, 0.824–0.877) in the development cohort and 0.851 (95% CI, 0.818–0.884) in validation cohort, and showed adequate calibration in both cohorts. The incidence of P-AMI infection increased steadily across risk score groups in both development and validation groups. The risk score also performed well in subgroup analysis for infection and in the outcomes of in-hospital all-cause death and major adverse cardiovascular events.
CONCLUSIONS The present risk score is easily applicable in clinical practice for identifying patients at high risk of infection and in-hospital outcomes in patients with STEMI undergoing PCI and may help clinical decision-making allowing for timely measures to improve clinical outcomes.
GW31-e0196
Boda Zhou, Ping Zhang
Beijing Tsinghua Changgung Hospital
OBJECTIVES Trimethylamine N-oxide (TMAO), a metabolite of gut microbes, has been reported to participate in the development of atherosclerosis and CVD. The purpose of our study was to assess whether the occurrence of in-stent restenosis (ISR) is associated with plasma TMAO level in acute coronary syndrome patients after drug eluting stent (DES) implantation.
METHODS This was a single retrospective case-control study performed in 1371 acute coronary syndrome (ACS) patients underwent percutaneous coronary intervention (PCI) and DES implantation. Of them, 15 symptomatic patients were found to suffer from ISR and included in this study (ISR group), 16 gender and age matched patients without ISR were included in this study (non-ISR group). High-performance liquid chromatography with tandem mass spectrometry was used to measure plasma TMAO levels.
RESULTS This study found no significant difference in plasma TMAO (247.40±181.30 ng/mL vs. 204.86±172.39 ng/mL, P=0.04) between ISR and non-ISR groups. Plasma TMAO level showed no significant correlation with ISR, but was significantly positively correlated with diabetes mellitus (r=0.45, P=0.01), serum HbA1c level (r=0.39, P=0.03) and serum creatinine level (r=0.47, P=0.007); significantly negatively correlated with female gender (r=–0.39, P=0.03). ISR was significantly positively correlated with diabetes mellitus (r=0.43, P=0.02), fasting blood glucose level (r=0.45, P=0.01), neutrophil to lymphocyte ratio (r=0.39, P=0.03) and syntax score (r=0.37, P=0.04); significantly negatively correlated with platelet (r=–0.39, P=0.03). Logistic regression analysis indicated that fasting blood glucose was the only independent predictors for ISR (B=0.669, 95% CI: 1.065–3.579, P=0.030).
CONCLUSIONS This study provides an information that plasma TMAO may not be related to ISR and plaque burden in ACS patients after DES implantation, while FBG may predict the development of ISR in these patients.
GW31-e0212
Ou Zhang, Yajun Xue, Ping Zhang
Beijing Tsinghua Changgung Hospital
OBJECTIVES To determine whether the pandemic of coronavirus disease 2019 (COVID-19) may affect clinical outcome of ST segment elevation myocardial infarction (STEMI) patients in North of Beijing, China, the epidemiological low risk area.
METHODS It was a single center retrospective observational study. Clinical characteristics and outcomes of all STEMI patients treated with primary percutaneous coronary intervention (PPCI) from January 24, 2020 to April 24, 2020 (group 2020) and in the same period in 2019 (group 2019) were compared.
RESULTS Totally 90 STEMI patients were included in our study (group 2020, n=51 vs. group 2019, n=39). No confirmed COVID-19 case in group 2020 tended to have longer door-to-balloon (DTB) time (135 (90, 184) vs. 76 (59, 102), P<0.0001), system delay time (from first medical contact to wire crossing time (206.5 (118, 313) vs. 92 (73, 114), P=0.003) and total delay time (from symptoms to wire crossing time) (333 (204, 581) vs. 173 (150, 362), P<0.0001)). There was no significant difference in the incidence of composite outcome of in-hospital death, cardiogenic shock, heart failure and use of mechanical circulatory support between group 2020 and 2019 (17 (33.33%) vs. 11 (28.21%), P=0.60). However, the rate of left ventricular aneurysm was significantly increased in group 2020 (27.45 vs. 5.13%, P=0.006) compared with 2019.
CONCLUSIONS COVID-19 may delay the treatment time of PPCI for STEMI patients, but no significant difference was observed on clinical outcome.
GW31-e0215
Yintang Wang1,2, Weihua Song1
1Chinese Academy of Medical Sciences and Peking Union Medical College, National Center for Cardiovascular Diseases, Fuwai Hospital
2Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University
OBJECTIVES Coronary artery ectasia (CAE) bears high risk of death and myocardial infarction. Risk stratification in CAE patients is crucial for their management, but there are no risk score systems intended for risk evaluation of CAE patients so far. This study is designed to investigate a user-friendly tool for risk evaluation, in order to help the clinical decision of the patients with CAE.
METHODS In a retrospective cohort of 595 hospitalized CAE patients from January 2009 and December 2013, the baseline characteristics (clinical history, biomarkers and quantitative coronary angiography variables) were collected. Follow-up via telephone interview were conducted by investigators blind for patients’ baseline characteristics and the endpoint event was the composite of all-cause death and non-fatal myocardial infarction. The candidate predictors of end-point event were analyzed using Cox proportional hazards regression models to derive a risk score in the form of nomogram. The predictive performance and discriminative ability of the novel nomogram were determined by concordance index (C-index) and calibration curve, that were validated internally with bootstraps resampling and leave-one-out cross-validation. According to the 60PthP and 90PthP pencentiles of the nomogram-derived score, the CAE patients were divided into three risk groups and risk stratification was further evaluated. The decision curve analysis (DCA) was conducted to evaluate clinical usefulness of the nomogram-assisted decisions to help improve CAE patients’ outcomes.
RESULTS During a median follow-up time of 62.3 (interquartile range 46.9–79.3) months, 26 all-cause deaths and 37 non-fatal myocardial infarctions were identified. In univariable analysis, age, BNP (Brain natriuretic peptide), hs-CRP (high sensitivity C-reactive protein), maximum dilated area of ectatic lesions, erythrocyte sedimentation rate, lymphocyte count, red blood cell count and hemoglobin were associated with the composite endpoint events. The final risk-prediction model named ABCD-CAE score included four items: age (A), BNP (B), hs-CRP (C) and maximum Dilated area of ectatic lesions (D). The nomogram yielded a C-index for end-point event of 0.72 (95% confidence interval, 0.64–0.79). The calibration curve demonstrated that there was good agreement between risk prediction by nomogram and actual observation of endpoint events. Compared with the low-risk group (score ≤100), the risk of composite events was significantly increased in the intermediate-risk group (score: 100–130) and high-risk group (score >130) [hazard ratio (95% confidence interval): 2.23 (1.23–4.06), P=0.008 and 7.02 (3.81–12.97), P<0.001 respectively]. Consistently, Kaplan-Meier curves were separated very well by the three groups both for composite events and all-cause death (log-rank test, both P<0.001). Compared with a model including clinical variables (age, sex, smoker, diabetes, hypertension, body mass index, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol ratio) alone, the ABCD-CAE model yielded a substantial net benefit when the decision thresholds approximately ranged from 25 to 65% in the DCA.
CONCLUSIONS The ABCD-CAE score is a simple four-item risk score, that provides a clinically useful tool for the risk prediction of all-cause death and myocardial infarction in patients with CAE. This user-friendly tool might support clinical decision making for the management of CAE.
GW31-e0217
Kokozheva Madina, Mekhman Mamedov
National Medical Research Center for Preventive Medicine, Moscow, Russia
OBJECTIVES Determination of the clinical course of CHD and risk factors in patients with diabetes mellitus.
METHODS A comparative clinical study included 200 patients with CHD (men and women aged 35–70 years). Depending on the glycemic status, patients were divided into two groups: with (n=100) and without (n=100) T2DM. All patients were questioned, conducted a clinical examination, instrumental studies, including ECG alone, ECHO KG, treadmill test and coronary angiography. The presence of concomitant diseases was also analyzed.
RESULTS In the group of people with diabetes, 40% smokes, and in the control group, the percentage of smokers was 60%. Alcohol abuse in the group of people with CHD and diabetes was 7%, and in the control group 10%. All patients in both groups had hypertension. In the group with diabetes, concomitant diseases were recorded 30% more compared to the group of people with coronary heart disease without diabetes and obesity, nephropathy, COPD and cerebrovascular diseases dominated. According to resting ECG and ECG, in both groups, most patients had left ventricular hypertrophy. The frequency of systolic dysfunction in patients with diabetes is detected by 50% more compared to the group without diabetes, and diastolic dysfunction is comparable and averages 80%. Among people with diabetes, stenosis of the LAD and proximal interventricular branch LCA are detected more often in comparison with patients without diabetes, while the frequency of stenosis up to 50 and 70% in both groups is comparable.
CONCLUSIONS Thus, a high frequency of risk factors is detected in both groups. In people with CHD and diabetes, the comorbidity of somatic diseases is detected more often due to nephropathy, cerebrovascular disease, COPD and obesity. Despite the comparable frequency of coronary artery stenosis in patients with diabetes mellitus, damage to some coronary arteries was more pronounced, which is associated with systolic dysfunction of the left ventricle.
GW31-e0218
Xue Wang, Jianjun Ruan, Heyu Meng, Lihong Li, Fanbo Meng
Department of Cardiology, The Third Hospital of Jilin University
OBJECTIVES To evaluate the correlation between the expression level of EGR1 gene in peripheral blood leukocytes and the risk of acute myocardial infarction (AMI); combined with the baseline data analysis of the study subjects, to explore the role of EGR1 gene in AMI.
METHODS In this study, we selected 86 patients with AMI as the case group and 77 patients with coronary heart disease as the control group. Both patients were diagnosed by coronary angiography. The clinical data of the two groups were analyzed and compared. In the peripheral venous blood, real-time fluorescence quantitative PCR was used to detect the mRNA expression of EGR1 gene in peripheral blood.
RESULTS The relative expression of EGR1 gene mRNA level in the group of patients with AMI and coronary heart disease showed that the relative expression of EGR1 gene mRNA in peripheral blood leukocytes of AMI group was significantly lower than that of control group, and its relative expression level was control group 0.49 times. The clinical data analysis results of the study objects indicated that there was no significant difference between the two groups in terms of age, smoking history, triglyceride level, total cholesterol level, low-density lipoprotein cholesterol level, high-density lipoprotein cholesterol level, hypertension diagnosis, type-II diabetes diagnosis. Further analysis of the relationship between clinical data and EGR1 gene expression results: the relative expression level of EGR1 gene mRNA level was independent of age (P=0.207), triglyceride levels (P=0.470), high density lipoprotein levels (P=0.949), low density lipoprotein levels (P=0.138); related to total cholesterol levels (P=0.017). Binary Logistic regression analysis showed that the low expression of mRNA in peripheral blood leukocytes of EGR1 gene is an independent risk factor for the development of coronary heart disease to AMI. Compared with the high expression of EGR1 gene, the risk of AMI with low expression of EGR1 gene is increased by 2.389 times.
CONCLUSIONS EGR1 gene is significantly under-expressed in peripheral blood of patients with AMI. Low expression of EGR1 gene is an independent risk factor for the occurrence of AMI. EGR1 gene may be used as a genetic marker to assess the risk of AMI.
GW31-e0238
Yuexi Wang, A. Rong
Department of Cardiology, 1st Hospital Affiliated to Inner Mongolian Medical University, Huhhot 010050, China
OBJECTIVES Indobufen as a new antiplatelet drug has been considered for antiplatelet replacement therapy before thrombolysis in patients with ST-segment elevation myocardial infarction who are intolerant of aspirin in the gastrointestinal tract. The aim of this trial was to investigate the efficacy and safety of antiplatelet therapy with indobufen in patients with troponin negative coronary artery stent implantation within one year compared with aspirin.
METHODS This trial is a prospective, open, non-inferiority, 1:1 randomized controlled exploratory study. We selected some patients with coronary artery disease (CAD) who underwent percutaneous coronary intervention (PCI) from June 2018 to February 2019 in the Department of Cardiovascular Medicine of our hospital. 60 patients who met the inclusion and exclusion criteria were selected and randomly assigned to the experimental group and the control group with 30 patients each. Within 24 hours after stent implantation, the experimental group received indobufen (100 mg bid) plus clopidogrel (75 mg qd), and the control group received aspirin (100 mg qd) plus clopidogrel (75 mg qd) dual antiplatelet therapy regularly for 12 months. The primary end point of net adverse clinical and cerebral events (NACCE)
RESULTS (1) The demographic data and baseline condition of the experimental group and the control group were balanced. There was no significant difference in the incidence of the end point events between the experimental group and the control group during the 12 month follow-up period. (2) The primary endpoint was analyzed by Kaplan-Meier method. The incidence of NACCE was 6.7% in the experimental group and 16.7% in the control group (HR, 0.238 [95% CI, 0.041–1.375]; Log-rank P=0.16). The choice of two dual antiplatelet regimens was not statistically associated with the occurrence of NACCE. (3) The incidence of major bleeding (BARC type 2, 3 and 5) at 12 months was 3.33% and 13.33% (HR, 0.238 [95% CI, 0.041–1.375]; Log-rank P=0.16) in the experimental group and the control group, and there was no statistical correlation between the two treatment regimens and the occurrence of major bleeding.
CONCLUSIONS Indobufen combined with clopidogrel as dual antiplatelet therapy did not significantly reduce the incidence of death, non-fatal myocardial infarction, ischemic stroke and major bleeding in troponin negative patients with CAD who received percutaneous coronary intervention and implanted at least one drug-eluting stent, but could play a stable antiplatelet role without increasing stent thrombosis Risk and occurrence of upper gastrointestinal adverse events.
GW31-e0239
Yuexi Wang, Nari Han
Department of Cardiology, Affiliated Hospital of Inner Mongolian Medical University, Huhhot 010050, China
OBJECTIVES Acute myocardial infarction (AMI) is one of the main causes of morbidity and mortality worldwide. The myocardial cells in patients with AMI are hypoxic and ischemic, and the structure and function of myocardium are changed. Some patients even eventually develop heart failure. Early diagnosis of AMI Treatment and prognosis play an important role. At present, clinical detection of troponin T (cTnT), creatine phosphokinase (CK), creatine phosphokinase isoenzyme (CK-MB) and myoglobin (MYO), the degree of increase is similar to the development and prognosis of the disease Related. Existing studies have confirmed that peripheral blood heat shock protein 70 (HSp70) is significantly abnormal in patients with acute myocardial infarction. Whether heat shock protein 70 can be used as a new diagnostic indicator for acute myocardial infarction needs further confirmation. Therefore, this study will explore the diagnostic significance of HSP70 in AMI.
METHODS This study selected 160 patients with acute myocardial infarction (male 112, female 48) in the Department of Cardiology, Inner Mongolia Medical University Affiliated Hospital from December 2017 to December 2018 and 40 healthy controls who were examined at the hospital’s physical examination center during the same period. Cases were divided into two groups: the Acute and convalescent phase. Serum heat shock protein 70 was detected by enzyme-linked immunosorbent assay (double antibody sandwich method) for statistical analysis.
RESULTS The serum HSP70 in the acute phase of the test group was significantly higher than that in the recovery period of the same group [25.37 (10.32, 50.08) μg/L vs. 0.088 (0.215, 0.557) μg/L and 0.092 (0.027, 0.976) μg/L, P<0.01]. The area under the ROC curve of the test group HSP70 was 0.722 (95% CI: 0.982–0.999), CK 0.883 (95% CI: 0.837–0.929), CK-MB 0.929 (95% CI: 0.894–0.964), cTNT 0.949 (95% CI: 0.918–0.980), BNP 0.853 (95% CI: 0.923–0.982), MY 0.775 (95% CI: 0.705–0.845). Serum HSP70 was positively correlated with CK, CK-MB, cTNT, MYO (P=0.0000). Logistic regression analysis showed that HSP70 was independently associated with the incidence of AMI (OR=3.201, 95% CI: 0.248–0.414).
CONCLUSIONS This study found that the serum HSP70 level of patients in the acute phase of the experimental group was significantly higher than that of the control group. Moreover, HSP70 levels were positively correlated with myocardial necrosis markers CK-MB and cTnT.
GW31-e0252
Yue Ma, Zhuoxuan Yang, Yue Zhou, Shubin Qiao
Department of Coronary Heart Disease, National Center for Cardiovascular Diseases, Fuwai Hospitai, CAMS & PUMC
OBJECTIVES Percutaneous coronary intervention (PCI) is the treatment of myocardial ischemia perfusion by using cardiac catheter technique to dredge the stenosis or occlusion of the coronary artery. There is a certain proportion of in-stent restenosis (ISR) after stent implantation that causes problems clinically. ISR involves many pathologic mechanisms, including intraplaque inflammation, lipid deposition, proliferation of vascular smooth muscle cells, endoluminal thrombus formation, and intraplaqu angiogenesis. Endothelins are 21-amino acid vasoconstricting peptides produced primarily in the endothelium having a key role in vascular homeostasis. Endothelin 1 (ET-1) is a potent vasoconstrictor that in patients with coronary artery disease is related with the occurrence of cardiovascular events. However, the low circulating concentration and the short half-life of ET-1 make it difficult to measure, big endothelin-1 (Big ET-1) is the precursor of ET-1 and has the same measurement value. The aim of this study is to investigate the relationship between big endothelin-1 and the risk of in-stent restenosis in patients with coronary artery disease undergoing PCI.
METHODS After retrieving the patient database, 3098 patients who diagnosed with coronary artery disease and underwent percutaneous coronary intervention (PCI) at Fuwai hospital from October 2014 to March 2015 were enrolled. All patients received standard dual therapy with aspirin 100 mg/day and clopidogrel 75 mg/day for 12 months following PCI. The patient’s level of plasma big endothelin-1 (Big ET-1), general condition, related risk factors of admission, other laboratory examination, coronary lesion characteristics, interventional therapy and other baseline data was recorded. All patients were followed up for the occurrence of in-stent restenosis (ISR) and other cardiovascular events. ISR was defined as the presence of >50% diameter stenosis in the stented segment. Statistical software SPSS22.0 was used for data analysis of the clinical epidemiological characteristics and prognosis. The study followed the principles outlined in the Declaration of Helsinki and it was approved by the ethics committee.
RESULTS In all participants, the level of plasma big endothelin-1 (Big ET-1) is associated with the risk of in-stent restenosis (ISR). Patients with a higher level of plasma Big ET-1 (>0.25 pmol/L) had a significantly higher incidence of ISR (2.89 vs. 0.44%, P=0.011) and incidence of angina after stent implantation (13.2 vs. 2.8%, P=0.007) during the follow-up. In logistic multivariable analysis, high level of plasma Big ET-1 (HR 1.968, 95% CI 1.083–5.022, P=0.031) was significantly associated with incidence of ISR.
CONCLUSIONS Elevated level of plasma big endothelin-1 is associated with the risk of in-stent restenosis in patients undergoing percutaneous coronary intervention.
GW31-e0253
Lihong Li, Xue Wang, Fanbo Meng
The Third Hospital of Jilin University
OBJECTIVES ZCCHC9 gene is a CCHC-type zinc-containing finger protein that has the function of binding DNA and RNA, and sometimes mediates protein interactions, and participates in various cellular processes in physiological conditions and diseases. This study aimed to assess whether the expression of ZCCHC9 gene can be used as a biomarker to predict the occurrence of acute myocardial infarction.
METHODS In this study, we selected 126 patients with acute myocardial infarction (AMI) as the experimental group, 117 patients with stable coronary artery disease (CAD) as the control group, collected peripheral venous blood, after total RNA extraction, cDNA synthesis, Real-time fluorescence quantitative PCR test to measure the expression level of ZCCHC9 gene at the mRNA level in peripheral blood.
RESULTS (1) The relative expression level of ZCCHC9 gene mRNA level in the objects showed that ZCCHC9 gene was differentially expressed between the experimental group and the control group. The relative expression level of ZCCHC9 gene mRNA level in the peripheral blood of AMI patients was significantly lower than that of the control group, and its relative expression was 0.753 times of the CAD group. (2) The analysis of the clinical date of the objects showed that there were no significant differences in age, gender, smoking history, serum lipid levels, previous history of hypertension, and diabetes between the AMI group and the stable CAD group. However, there was a significant difference in the fasting blood glucose levels between the two groups (Z=-3.168, P=0.002). (3) The analysis of whether there was correlation between clinical date and the relative expression level of ZCCHC9 gene mRNA level showed that the relative expression level of ZCCHC9 gene mRNA level was no related to age, serum triglyceride, total cholesterol, high density lipoprotein, low density lipoprotein and the fasting blood glucose. (4) Binary Logistic regression analysis showed that the low expression of ZCCHC9 gene is an independent risk factor of AMI. Compared with the high expression of ZCCHC9 gene, the risk of AMI in the low expression of ZCCHC9 gene group was increased by 2.597 times. The low expression of ZCCHC9 gene in peripheral blood can be a standard to diagnose AMI. And its sensitivity was 27.4%, its specificity was 88.9%. (5) The relationship between the relative expression level of ZCCHC9 gene mRNA level and concentration of serum troponin I showed that the relative expression level of ZCCHC9 gene mRNA level had no correlation with serum troponin I concentration (P>0.05), that is, the relative expression level of ZCCHC9 gene may not predict the size of AMI.
CONCLUSIONS The expression of ZCCHC9 gene in peripheral blood of patients with AMI was significantly lower than that of patients with stable CAD. The low expression of ZCCHC9 gene is an independent risk factor for AMI. The low expression of ZCCHC9 gene in peripheral blood may used as a genetic marker to predict the occurrence of AMI.
GW31-e0254
Lihong Li, Xue Wang, Fanbo Meng
The Third Hospital of Jilin University
OBJECTIVES NUMB is an endocytic adaptor protein that has the function of controlling the physiological development process and plays a key role in the occurrence of diseases. To evaluate whether the low expression of NUMB gene in peripheral blood can be used as a molecular marker to predict the early diagnosis of acute myocardial infarction.
METHODS In our study, we selected 124 patients with acute myocardial infarction (AMI) as the experimental group, and 115 patients with stable coronary artery disease (CAD) as the control group, collected their peripheral venous blood, after total RNA extraction, cDNA synthesis, real-time fluorescence quantitative PCR test was used to measure the expression level of NUMB gene at mRNA level in peripheral blood.
RESULTS (1) The relative expression level of NUMB gene mRNA level of objects indicated that NUMB gene was differentially expressed between the experimental group and the control group, the relative expression level of NUMB gene in peripheral blood of AMI patients was significantly lower than that of the control group, and its relative expression quantity was 0.932 times of that in the control group. (2) The analysis of the clinical date of objects showed that there were no significant statistical differences between the two groups in terms of gender, history of hypertension, smoking history, diabetes, serum triglycerides, total cholesterol, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol. patients in the AMI group were significantly older than the control group (t=-2.318, P=0.020), and there was a significantly difference in the fasting blood glucose between the two groups (z=-2.505, P=0.012). (3) The analysis of whether there was correlation between clinical data and the relative expression level of NUMB gene mRMA level showed that the expression level of NUMB gene mRNA level was no related to the fasting blood glucose (P=0.551), and it was no related to age (P=0.645). There was no correlation between the expression of NUMB gene and fasting blood glucose and age. (4) Binary Logistic regression analysis indicated that low expression of NUMB gene is an independent risk factor of AMI. Compared with the high expression of NUMB gene group, the risk of AMI in the low expression of NUMB gene group was increased by 3.287 times. Advanced age was also an independent risk factor of AMI, which can increase the risk of AMI by 1.853 times. Fasting blood glucose was not an independent risk factor of AMI. (5) The relationship between the relative expression of NUMB gene and serum concentration of troponin I showed that there was no correlation between the expression of NUMB gene in peripheral blood and serum troponin I concentration (r=-0.027, P=0.797), that is, the expression level of NUMB gene in peripheral blood may not predict the size of AMI.
CONCLUSIONS The expression of NUMB gene in peripheral blood of patients with AMI was significantly lower than that of patients with stable CAD. Patients with low expression of NUMB gene in peripheral blood are more likely to develop AMI. The low expression of NUMB gene is an independent risk factor of AMI. The low expression of NUMB gene in peripheral blood may become a molecular marker for early diagnosis of AMI.
GW31-e0256
Yining Dai, PengCheng He, Ning Tan, YuanHui Liu
Department of Cardiology, Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Coronary Heart Disease Prevention, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences
OBJECTIVES Post-contrast acute kidney injury (PC-AKI) is associated with a prolonged hospital stay and increased mortality. PC-AKI cannot be treated effectively after it occurs; thus, avoiding this condition is particularly important. Statins have been shown to reduce the risk of PC-AKI in patients undergoing percutaneous coronary intervention (PCI). However, the preventive effect of rosuvastatin versus atorvastatin on PC-AKI in patients with ST-segment elevation myocardial infarction (STEMI) undergoing PCI remains unclear.
METHODS Patients with STEMI undergoing PCI between January 2010 and May 2016 were consecutively enrolled. Finally, 1300 patients were included and divided into two groups according to the statin type (atorvastatin: n=1040; rosuvastatin: n=260). All patients were administrated with statins at admission. Patients in the two groups received statin therapy (rosuvastatin: 10 mg daily; atorvastatin: 20 mg daily) before contrast agent exposure. The primary endpoint was PC-AKI defined as an absolute increase of ≥0.5 mg/dL in the level of serum creatinine or an increase of ≥25% over baseline within 48–72 hours after contrast media exposure.
RESULTS In total, 245 (18.8%) patients developed PC-AKI. The atorvastatin and rosuvastatin groups had similar rates of PC-AKI (19.1 vs. 17.7%, P=0.595), in-hospital mortality (4.1 vs. 3.8%, P=0.833), and major adverse clinical events (MACE). Multivariate logistic regression analysis revealed that rosuvastatin treatment had an effect similar to atorvastatin regarding PC-AKI (odds ratio [OR]=0.97, 95% confidence interval [CI], 0.66–1.43, P=0.874). Propensity score analyses and subgroup analysis demonstrated similar results for PC-AKI. Kaplan-Meier survival curves and Cox proportional regression showed that the atorvastatin and rosuvastatin groups had no differences regarding follow-up mortality.
CONCLUSIONS Rosuvastatin exerted a similar preventive effect against PC-AKI and showed similar levels of in-hospital and follow-up all-cause mortality and in-hospital MACE compared with atorvastatin in patients with STEMI undergoing PCI.
GW31-e0259
Lianna Xie, Xianjing Wei, Zezhou Xie, Shengying Jia, Siwei Xu, Chunlin Jiang, Xingping Xiao, Botao Luan, Kaijun Wang, Xue Yang
Affiliate Zhongshan Hospital of Dalian University
OBJECTIVES Asymptomatic radial artery occlusion remains the most common complication in transradial coronary interventional procedure. To prevent radial artery occlusion, distal radial access site has been suggested recently. We aimed to describe our experience and to assess feasibility and safety of this new access site for coronary angiography (CAG) and percutaneous coronary intervention (PCI).
METHODS One thousand and sixty three patients were assigned to undergo CAG or procedural PCI through distal radial access between 1 January 2018 and 31 December 2019. The size of radial sheath used was 5 or 6 French (F). The sheath was removed at procedure termination, and hemostasis was obtained by compression bandage with gauze.
RESULTS The successful rate of radial artery cannulation via distal radial access was 89.7%. Mean age of successful cases is 64.6±11.2 years (26–94 years) with 35.6% women. 38.1% of procedures were PCI. 28.2% of procedures were via left distal radial access. 6F sheath was used in 63.2% of cases. Hemostasis was obtained within 2 hours in 89.5% patients. There were 110 procedural failures: artery puncture failed in 59 cases, wire failed in 49 cases, and sheath failed in 2 cases. Complications potentially related to distal radial access were as follows: radial artery occlusion at the access site 1.4%, forearm radial artery occlusion 0.4%, hematoma of forearm 0.5%, and transient thumb numbness 0.2%.
CONCLUSIONS Distal radial access is a feasible and safe access and can be used as a rational alternative to traditional radial access for coronary interventional procedure.
GW31-e0279
N.F. Tashkenbayeva, D.A. Alimova, R.K. Trigulova, F.M. Bekmetova, L.T. Ilkhamova
Republican specialized scientific Practical Medical Center of Cardiology of the Uzbekistan Public Health Ministry
OBJECTIVES To assess glucagon-like peptide-1 Liraglutide (LG) efficacy on glycemia control, losing weight, lipid profile, and EchoCG parameters in patients with CHD and diabetes mellitus type 2 (DM 2).
METHODS The study included 15 patients (LG) unstable angina UA (ESC) and diabetes mellitus 2 (DM-2) (WHO, 1999) in the age 56.4±2.7 years old. Patients BMI was 30.7±0.6 kg/m2; duration of DM 2 was 8.1±2.1 years. Using standard methods we determined complete lipid spectrum, fasting glycemia (FG), postprandial glycemia (PG), HbA1c, body weight, and EchoCG parameters. All Echocardiograms were performed by one cardiologist using ultra sound «En VisorC» system («PHILIPS», Holland) with 3.5 MHz sensor with a patient in lying position on the left flank with 45° angle in compliance with standard methods. Therapy mode: anticoagulants, antiplatelet agents, nitrates, beta-blockers, RAAS blockers, statins, Liraglutide. Liraglutide’s efficacy was assessed according to the decrease of HbA1c level compared to original HbA1c ≥0.5%. A group of patients with CHD with DM 2, who administered Gliclazide (GL) with parameters corresponding to the studied parameters of the basic group was considered to be a control group (n–10). Follow-up period was 3 months.
RESULTS Within the follow-up period there was a notable improvement of HbA1c compared to original level in both groups: LG from 9.2±1.6 to 7.3±1.1, P=0.004; and GL 8.9±1.5, P=0.04. However, in relation to fasting glycemia significant improvement of glycemic control was observed in the 8th group with LG: LG from 10.5±3.1 to 8.1±2.4 mmol/L, P=0.07; and GL 10.9±4.3 to 8.9±3.8 mmol/L, P=0.22. It was probably linked with different number of patients in two groups. Alterations of BMI from the original level till the 12th week was equal to -1.2 kg (P=0.007) in LG group and -0.2 kg (P=0.9). In relation to dynamic lipid profile parameters with background 3-months therapy there was insignificant tendency for decrease, and particularly dynamic LDL: LG group to (-) 4.1±3.6 mg/dL (P=0.6) versus GL group (-) 0.7±1.5 mg/dL (P=0.9); HDL (-) 0.3±2.1 versus 0.6±1.9 mg/dL (P=0.8) respectively; fasting triglycerides (-) 11.6±3.7 (P=0.19) versus (-) 8.1±4.7 (P=0.8) mg/dL. In LG group basic level ten patients had diastolic dysfunction (DD) I stage, while 5 patients (33.3%) had DD II stage. By the 3rd month of therapy in LG group there was observable improvement of diastolic function and transfer of all patients from DD II stage to the group of patients with DD I stage. None of the patients stopped Liraglutide therapy and informed of hypoglycemia. Clinical tolerance of the agent was good.
CONCLUSIONS In clinical practice addition of Liraglutide to the therapy of patients with CHD and DM type 2 with insufficient control of carbohydrate profile improves glycemic control, diminishes body weight, does not affect lipid exchange parameters with background basic therapy. In this preliminary research Liraglutide had a favorable effect on DD in patients with DM type 2. It is an important conclusion, as DD is a known additive risk of unfavorable events development.
GW31-e0280
D.A. Alimova, N.F. Tashkenbayeva, R.Kh. Trigulova, F.M. Bekmetova, M.A. Musayeva
Republican Specialized Scientific Practical Medical Center of Cardiology of the Uzbekistan Public Health Ministry
OBJECTIVES To assess DDLV status dependent on the markers determining cardiovascular events development risks.
METHODS We studied forty patients with IHD, who applied to the 2nd and 6th units of the RSSPMC of Cardiology of the MH of the RUz in destabilization period of angina combined with DM type 2 and hypertonic disease (HD) aged 59.2±9.5 years old. Using standard methods we determined complete lipid spectrum (TC, CL LDL, VLDL, HDL, TG), fasting glycemia (FG), postprandial glycemia (PG), HbA1c, and C-reactive protein (CRP). Therapy mode: anticoagulants, antiplatelet agents, nitrates, RAAS blockers, beta-blockers, and statins. Patients with DM type 2 administered metformin, and sulphonyl urea agents. Exclusion criteria were heart failure (HF) II B, chronic kidney disease (CKD) 4, any incretin therapy, and insulin therapy. At the 3–5th day after status stabilization with basic therapy 30 patients were randomly selected for administration of sitagliptin/metformin (SG/M) with a day dose 50/500–50/1000 mg/day, with monitoring of glycemia, heartbeat rate, SAP, DAP, lipid profile, and CRP level. The rest 10 patients continued taking gliclazide (GL). Primary results were changes in ratio of e′ speed and E/e′ from the original level within 24 weeks (6 months) follow-up. Secondary results included: HbA1c, lipid profile. CRP level. Follow-up duration was 6 months.
RESULTS Decrease of HbA1c and body weight was significantly higher in SG/M group, than in GL group (-0.8±0.5% versus -0.4±0.5, P<0.005; -1.7±3.9 kg versus 0.5±3.1 kg, P<0.05 respectively). In the lipid profile and CRP level of the compared groups there were no changes. We did not reveal any significant alterations in the speed (e′) and E/e′ of the original level within the whole follow-up period in any of the groups.
CONCLUSIONS Sitagliptin efficacy in relation to glycemic status was more expressed compared to Gliclazide (decrease of HbA1c level). In echocardiographic parameters of diastolic function of the left ventricle in patients with CHD and DM 2 there were yet no observable differences.
GW31-e0286
Li Gang, Guo Yifang, Zuo Qingjuan, He Lili, Wang Yan, Zhang Lu, Zhang Tingting, Wang Qiuyan, Liang Yi
Hebei General Hospital
OBJECTIVES Chronic renal dysfunction and coronary heart disease are the major diseases that affect the health of the elderly. The change of left ventricular structure and function is one of the most prominent features of heart damage in chronic kidney disease patients. Serum sulfatide, a very sensitive indicator of atherosclerosis and changes in renal function, may be important in monitoring the occurrence and progression of cardiac and renal insufficiency. This study aimed to investigate the relationship between serum sulfatide and glomerular filtration rate in patients with coronary heart disease and left ventricular structural and functional changes in order to further understand the inherent relationship between renal dysfunction and cardiovascular events.
METHODS Sixty hospitalized patients diagnosed with coronary heart disease were enrolled in this research. All of them were assigned to 3 groups according to the tertiles of the estimated glomerular filtration rate (eGFR): A group contain 19 patients (eGFR<57.28 mL min-1 1.73 m-1), B group contain 21 patients (57.28<eGFR<74.86 mL min-1 1.73 m-1), C group contain 20 patients (eGFR>74.86 mL min-1 1.73 m-1). Venous blood was collected early in the morning and sent to the laboratory for detection of serum creatinine. The eGFR was calculated according to the simplified MDRD formula. Another 3 mL of venous blood was collected and the serum was separated. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry was used to detect the level of serum sulfatide. Left ventricular structure and function were measured by echocardiography. Relevant test indicators were compared between different renal function patients. Then, the correlation between serum sulfatide concentration, eGFR and left ventride structure and function were analyzed.
RESULTS (1) With the decline of eGFR, serum sulfatide levels gradually decrease (A<B<C, P<0.05), and serum creatinine levels gradually increased (A>B>C, P<0.05). When the eGFR is significantly decreased, the left atrial diameter increases (A>C, P<0.05), the ejection fraction decreases (A<B, P<0.05), and the left ventricular end-systolic diameter increases (A>C, P<0.05). (2) Serum sulfatide concentration was negatively related to left atrial diameter (r=-0.635, P<0.05), or E peak (r=-0.577, P<0.05), but was significantly positively related to eGFR (r=0.817, P<0.01). (3) eGFR was significantly negatively correlated with left atrial diameter and E peak (r=-0.716, P<0.05; r=-0.699, P<0.05, respectively). After multivariable adjustments, serum sulfatide is still significantly correlated with eGFR (P<0.05), and eGFR is significantly negatively correlated with E peak (P<0.05).
CONCLUSIONS Serum sulfatide and eGFR levels were negatively correlated with left atrium diameter. Compared with group C, group A had a gradually increased left atrial diameter as the level of serum sulfatide and eGFR decreased. B group compared with A group, serum sulfatide and glomerular filtration rate decreased significantly, can cause the ejection fraction decreased. Due to the enlargement of left atrium and the decline of the ejection fraction, it is an important predictor of the prognosis of atrial fibrillation, cardiac insufficiency and cardiovascular disease, suggesting that the joint monitoring of serum levels of sulfatide and eGFR may open up a new idea for delaying the progression of cardiac insufficiency.
GW31-e0362
Dili Xie1, Yong Chen2
1Geriatric Cardiovascular Department, Sichuan Academy of Medical Science & Sichuan Provincial People’s Hospital
2Cardiovascular Departmen, Sichuan Academy of Medical Science & Sichuan Provincial People’s Hospital
OBJECTIVES This study was to compare the efficacy and safet among three different antithrombotic therapies in elderly post-PCI patients with atrial fibrillation (AF).
METHODS One hundred and forty Chinese cases with chronic coronary syndromes (CCS), permanent non-valvular AF, and diabetes mellitus (DM), were included in this study. The age of patients ranged from 70 to 78 years old. All of them received PCI therapy. After PCI, three different antithrombotic therapies were adopted in the patients. Ticagrelor dual therapy group first received one-year treatment with rivaroxaban (15 mg o.d.) and ticagrelor (90 mg bid). Rivaroxaban triple therapy group first received three-month triple therapy with rivaroxaban (15 mg o.d.), clopidogrel (75 mg o.d.) and aspirin (100 mg o.d.), then received nine-month dual therapy with rivaroxaban (15 mg o.d.) and clopidogre (75 mg o.d.). Warfarin triple therapy group first received three-month therapy with warfarin (INR 2.0–2.5, TTR>70%), clopidogrel (75 mg o.d.) and aspirin (100 mg o.d.), then received nine-month dual therapy with warfarin (INR 2.0–3.0, TTR>70%) and clopidogrel (75 mg o.d.). All the patients received rabeprazole (10 mg o.d.) during antithrombotic therapies after PCI. The incidences of cardiovascular events and hemorrhagic events were analyzed among the three groups.
RESULTS There were no differences of age, gender, BMI and CHA2DS2-VASC score among the three groups, but HAS-BLED score, proportion of high bleeding risk cases, proportion of frail patients in the warfarin triple therapy group was lower than in the other two groups (P<0.05). The incidences of stroke/TIA, non-central nervous system (CNS) embolism, myocardial infarction (MI) and unstable angina in the ticagrelor dual therapy group and in the rivaroxaban triple therapy group were lower than in the warfarin triple therapy group (P<0.05), however, there was no statistical difference of the cardiovascular events between the two groups. The incidences of gastrointestinal, intracranial, urinary tract bleeding in the ticagrelor dual therapy group were lower than in the other two groups (P<0.05). Each group had no hemoptysis case. There was no case with severe bleeding in the ticagrelor dual therapy group.
CONCLUSIONS Dual antithrombotic therapy with ticagrelor and rivaroxaban was not inferior to triple antithrombotic therapy with rivaroxaban, clopidogrel and aspirin, and was superior to triple antithrombotic therapy with warfarin, clopidogrel and aspirin, to prevent cardiovascular events. Besides the dual therapy can also reduce the bleeding risk significantly. Therefore, this dual antithrombotic therapy maybe a good choice in elderly post-PCI patients with CCS and AF.
GW31-e0369
Jianfeng Zheng1, TingTing Guo1, Yuan Tian2, Yong Wang1, XiaoYing Hu1, Yue Chang1, Hong Qiu1, KeFei Dou1, YiDa Tang1, JinQing Yuan1, YongJian Wu1, HongBing Yan1, ShuBin Qiao1, Bo Xu1, YueJin Yang1, RunLin Gao1
1Fu Wai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
2Urumqi Friendship Hospital, Urumqi, Xinjiang Uygur Autonomous Region, China
OBJECTIVES The mechanism and characteristics between early and late drug-eluting stent in-stent restenosis (DES-ISR) have not been fully clarified. Whether there are different outcomes among those patients being irrespective of their repeated treatments remain a knowledge gap.
METHODS A total of 250 patients who underwent initial stent implantation in our hospital, and then were readmitted to receive treatment for the reason of recurrent significant DES-ISR in 2016 were involved. The patients were categorized as early ISR (<12 months; E-ISR; n=32) and late ISR (≥12 months; L-ISR; n=218). Associations between patient characteristics and clinical performance, as well as clinical outcomes after repeated percutaneous coronary intervention (PCI) were evaluated. Primary composite endpoint of major adverse cardiac events (MACE) included cardiac death, non-fatal myocardial infarction, or target lesion revascularization (TLR).
RESULTS Most baseline characteristics are similar in both groups, except for the period of ISR, initial pre-procedure TIMI, and some serum biochemical indicators. The incidence of MACE (37.5 vs. 5.5%; P<0.001) and TLR (37.5 vs. 5.0%; P<0.001) is higher in the E-ISR group. After multivariate analysis, E-ISR (odds ratio [OR], 13.267; [95% CI 4.984–35.311]; P<0.001) and left ventricular systolic dysfunction (odds ratio [OR], 6.317; [95% CI 1.145–34.843]; P=0.034) are the independent predictors for MACE among DES-ISR patients in the mid-term follow up of 12 months.
CONCLUSIONS Early ISR and left ventricular systolic dysfunction are associated with MACE during the mid-term follow-up period for DES-ISR patients. The results may benefit the risk stratification and secondary prevention for DES-ISR patients in clinical practice.
GW31-e0387
Haifeng Zhang1,2, Qingyuan Gao1,2, Chengzhang Xu1,2, Yangxin Chen1,2, Shaohua Wang1,2, Zhiteng Chen1,2, Jingfeng Wang1,2
1Department of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510120, PRC
2Laboratory of Cardiac Electrophysiology and Arrhythmia in Guangdong Province, Guangdong 5101120, PRC
OBJECTIVES The CD14 gene C-260T polymorphism has been reported to be associated with ischemic heart disease, but results were conflicting. To evaluate the role of the CD14 C-260T polymorphism in ischemic heart disease, we performed meta-analyses of all available data.
METHODS Comprehensive searches for studies on the association between the genotypes (CC, CT, TT) distributions and ischemic heart disease risk were performed. Patients with acute coronary syndrome, prior myocardial infarction, stable angina pectoris, or angiographic coronary artery stenosis were included. Potential sources of heterogeneity were explored by meta-regression. Analyses were performed under European, East Asian, and Indian studies, respectively.
RESULTS Data were available for 19 studies involving 11,813 cases and 6196 controls. The summary odds ratio under the recessive model was 1.53 (95% confidence interval: 1.20–1.96) for East Asian studies published in English language journals on overall ischemic heart disease. Pooled odds ratios under the codominant model were about 1.81 (95% confidence interval: 1.36–2.40) and 1.70 (95% confidence interval: 1.26–2.29) for Chinese studies on overall ischemic heart disease and other ischemic heart diseases (angina pectoris and angiographic coronary artery stenosis), respectively. No significant association was found in a European population, an Indian population, or the vulnerable plaque ischemic heart disease (acute coronary syndrome and prior myocardial infarction) subgroup of an East Asian population.
CONCLUSIONS It is probable that T allele and TT genotype are associated with ischemic heart disease in the East Asian population but not in the European or Indian populations. Further studies are warranted to assess these associations in greater detail, especially in East Asian and Indian populations.
GW31-e0393
Xiaomin Tian, Heyu Meng, Fanbo Meng
China-Japan Union Hospital of Jilin University
OBJECTIVES Sub1, also known as p15, PC4, P14 (hereinafter referred to as PC4), is commonly found in 27 tissues of the human body, and most expressed in brain and fat tissue. Sub1 can rapidly collect DNA damage sites, combine with DNA damage sites early and briefly, play a role in detecting and/or exposing DNA damage, specifically protect DNA from oxidative damage, and reduce chronic inflammation. Inflammation plays an important role in the development of myocardial infarction. The purpose of this study is to evaluate whether the expression level of sub1 gene in peripheral blood can be used as a biomarker to predict the risk of acute myocardial infarction (AMI).
METHODS The peripheral blood of 75 patients with stable coronary heart disease and 113 patients with acute myocardial infarction were collected. The expression of sub1 gene was detected by real-time fluorescence quantitative polymerase chain reaction (PCR) and Western blot analysis.
RESULTS The results showed that the expression level of sub1 gene in the peripheral blood of patients with AMI was significantly lower than that of patients with stable coronary heart disease (z=−2.095, P 0.036). The low expression of sub1 gene had no correlation with blood lipid, blood glucose, smoking history, age and blood pressure.
CONCLUSIONS The expression of sub1 gene in the peripheral blood of patients with acute myocardial infarction was significantly lower than that of patients with stable coronary heart disease. Its low expression was an independent risk factor and could be used as a biomarker to predict acute myocardial infarction.
GW31-e0400
Fenghua Song1,2, Chaojian Zhang1,2
1Department of Cardiology, First Affiliated Hospital of Zhengzhou University
2Key Laboratory of Cardiac Injury and Repair of Henan Province, Zhengzhou, China
OBJECTIVES In present study, we aimed to explore the association of MPVLR with long-term mortality in CAD patients with diabetes mellitus (DM) who underwent percutaneous coronary intervention (PCI).
METHODS A total of 838 patients with CAD and DM who underwent PCI in the department of cardiology, the First Affiliated Hospital of Zhengzhou University from January 2013 to December 2017 were followed up. According to the cutoff value of MPVLR, all of the patients were divided into two groups: the low-MPVLR group (<5.69, n=583) and the high-MPVLR group (≥5.69, n=240). Multivariate Cox proportional hazards regression model was used to detect the predictors of endpoint events.
RESULTS In univariate analysis, there was significant difference in cardiovascular mortality (CM) (P<0.001) and all-cause mortality (ACM) (P<0.001) between the two groups. Multivariate Cox regression analysis showed that the MPVLR was an independent risk factor for long-term CM (hazard risk [HR]=3.579, 95% confidence interval [CI]: 1.602–8.078, P=0.002) and ACM (HR=2.390, 95% CI: 1.278–4.469, P=0.006) in patients with CAD and DM who underwent PCI.
CONCLUSIONS Elevated MPVLR was an independent and reliable predictor of long-term mortality patients with DM who underwent PCI.
GW31-e0402
Xue Wang, Lihong Li, Fanbo Meng
The Third Hospital of Jilin University
OBJECTIVES To evaluate the correlation between the expression level of ADIPOR1 gene in peripheral blood leukocytes and the risk of acute myocardial infarction (AMI); combined with the baseline data analysis of the study subjects, to explore the role of ADIPOR1 gene in AMI.
METHODS In this study, we selected 69 patients with AMI as the case group and 63 patients with non-coronary heart disease as the control group. Both patients were diagnosed by coronary angiography. The clinical data of the two groups were analyzed and compared. In the peripheral venous blood, real-time fluorescence quantitative PCR was used to detect the mRNA expression of ADIPOR1 gene in peripheral blood.
RESULTS The relative expression of ADIPOR1 gene mRNA level in the group of patients with AMI and non-coronary heart disease showed that the relative expression of ADIPOR1 gene mRNA in peripheral blood leukocytes of AMI group was significantly lower than that of control group, and its relative expression level was control group 0.49 times. The clinical data analysis results of the study objects indicated that there was no significant difference between the two groups in terms of smoking history, total cholesterol level, low-density lipoprotein cholesterol level, high-density lipoprotein cholesterol level, hypertension diagnosis. However, there were significant differences in age, history of type 2 diabetes, smoking history, and triglyceride levels, and the differences were statistically significant (P<0.05). Further analysis of the relationship between clinical data and ADIPOR1 gene expression results: the relative expression level of ADIPOR1 gene mRNA level was independent of age (P=0.76), triglyceride levels (P=0.77), high density lipoprotein levels (P=0.99), low density lipoprotein levels (P=0.73), history of type 2 diabetes (P=0.80), and total cholesterol levels (P=0.42). Binary Logistic regression analysis showed that the low expression of mRNA in peripheral blood leukocytes of ADIPOR1 gene is an independent risk factor for the development of AMI. Compared with the high expression of ADIPOR1 gene, the risk of AMI with low expression of ADIPOR1 gene is increased by 3.01 times.
CONCLUSIONS ADIPOR1 gene is significantly under-expressed in peripheral blood of patients with AMI. Low expression of ADIPOR1 gene is an independent risk factor for the occurrence of AMI. ADIPOR1 gene may be used as a genetic marker to assess the risk of AMI.
GW31-e0406
Zhenyu Xiong, Xiangbin Zhong, Xiaodong Zhuang, Shaozhao Zhang, Xinxue Liao
Department of Cardiology, the First Affiliated Hospital, Sun Yat-Sen University
OBJECTIVES To measure the association between LP(a) and coronary artery calcification (CAC) during young to middle adulthood.
METHODS The Coronary Artery Risk Development in Young Adults study enrolled 5115 healthy black and white American aged 18–30 years at baseline. LP(a) was measured by a double monoclonal antibody enzyme-linked immunosorbent assay method at year 5. CAC was identified by computed tomography at years 15, 20 and 25.
RESULTS Of 3146 participants, the average (SD) level of LP (a) was 20.6 (21.5). About 28.2% participants developed CAC during 25 years. After multivariable adjustment, the hazard ratio for CAC associated with LP (a) was 1.11 (95% CI 1.01, 1.23) per 5 years increment. Results were similar when categorized individuals by clinical practice: compared with LP (a) ≤30 mg/dL, the hazard ratio of LP (a) >50 mg/dL was 1.26 (1.03, 1.56).
CONCLUSIONS Severe LP (a) level during young adulthood is independently associated with CAC at midlife, suggesting its potential use in predicting cardiovascular disease in an early stage.
GW31-e0409
Wang Jun1, Yang Yi2, Haibing Jiang2
1The People’s Hospital of Xuancheng City
2The Xinjiang Medical University Affiliated Hospital of Traditional Chinese Medicine
OBJECTIVES Inflammation is involved in the pathogenesis and progression of coronary artery diseases (CADs), including acute coronary syndrome. The neutrophil-to-lymphocyte ratio (NLR) has been identified as a novel marker of the pro-inflammatory state. We aimed to evaluate the predictive efficacy of the NLR for the prognosis of patients with new-onset ACS.
METHODS We retrospectively included consecutive patients with new-onset ACS treated with emergency coronary angiography. NLR was measured at baseline and analyzed by tertiles. The severity of coronary lesions was evaluated by the Gensini score. Correlations of NLR with the severity of CAD and the incidence of major adverse cardiovascular diseases (MACEs) during follow-up were determined.
RESULTS Overall, 737 patients were included. The NLR was positively correlated with the severity of coronary lesions as assessed by Gensini score (P<0.05). During the follow-up period (mean, 43.49±23.97 months), 65 MACEs occurred. No significant association was detected between baseline NLR and the risk of MACEs during follow-up by either Kaplan–Meier or Cox regression analysis. The results of multivariable logistic regression analysis showed that a higher NLR was independently associated with coronary lesion severity as measured by the GS (1st tertile vs. 3rd tertile hazard ratio [HR]: 0.527, P<0.001, and 2nd tertile vs. 3rd tertile HR: 0.474, P=0.025).
CONCLUSIONS The NLR may be associated with coronary disease severity at baseline but is not associated with adverse outcomes for patients with new-onset ACS.
GW31-e0410
Jun Wang1, Yang Yi2, Benfang Wang3
1The People’s Hospital of Xuancheng City, Anhui 242000, China
2Department of Cardiology fourth ward, The Xinjiang Medical University Affiliated Hospital of Traditional Chinese Medicine, Urumqi 830011, China
3Department of Coronary Heart Disease, The First Affiliated Hospital of Bengbu Medical College; Anhui 233000, China
OBJECTIVES Right bundle-branch block (RBBB) and left bundle-branch block (LBBB) plays a role in the pathogenesis and progression of coronary artery disease (CAD); however, the effect of RBBB and LBBB on the severity of coronary artery is not well characterized.
METHODS We retrospectively analyzed data pertaining to consecutive patients with RBBB or LBBB who underwent coronary angiography at two centers. The severity of coronary lesions was evaluated using the SYNTAX score. The differential effect of new-onset RBBB, old RBBB, new-onset LBBB, and old LBBB on the severity of CAD and its association with clinical characteristics was quantified. Multivariate logistic regression analysis was performed to evaluate the effect of RBBB and LBBB on the degree of coronary atherosclerosis.
RESULTS Out of the 243 patients, 72 patients had old LBBB, 37 had new-onset LBBB, 93 patients had old RBBB, and 41 patients had new-onset RBBB. On univariate analysis, age, systolic blood pressure, diastolic blood pressure, creatinine, serum glucose, and glycosylated hemoglobin level were associated with high SYNTAX score (P<0.05 for all). Patients in the new-onset RBBB, old RBBB, new-onset LBBB, and old LBBB groups showed significant differences in baseline characteristics and coronary atherosclerosis (P<0.05 for all). However, there were no significant between-group differences with respect to the degree of coronary atherosclerosis as assessed by SYNTAX score. On multivariate logistic analysis, age, systolic pressure, diastolic blood pressure, creatinine, serum glucose, lipoprotein(a), and previous myocardial infarction were associated with high SYNTAX score (P<0.05).
CONCLUSIONS New-onset RBBB, old RBBB, new-onset LBBB, and old LBBB were not associated with the severity of coronary lesions as assessed by SYNTAX score. Our findings suggest the clinical significance of RBBB in patients with chest pain due to suspected CAD.
GW31-e0419
Xinliang Zou1, Li Nie2, Yi Liao3, Zhihui Liu1, Xiang Xu1, Haoran Qin1, Haidong Wang3, Jianping Liu1, Guoxiang He1, Tao Jing1
1Department of Cardiology, The First Hospital Affiliated to Army Medical University (Southwest Hospital)
2Department of Internal Medicine, Central Hospital of Wandong
3Department of Thoracic Surgery, The First Hospital Affiliated to Army Medical University (Southwest Hospital)
OBJECTIVES Patients with nephrotic syndrome often have disorders of lipid metabolism. Elevated circulating low-density lipoprotein cholesterol (LDL-C) increases the risk of Atherosclerosis and is a risk factor for nephrotic syndrome complicated with cardiovascular events. Statins are the cornerstone of lipid management, but it is unclear whether statins can effectively reduce the occurrence of cardiovascular disease and improve the prognosis of patients with nephrotic syndrome who do not meet the criteria for chronic kidney disease. We designed a retrospective nested case-control study to observe whether the treatment of statins can reduce cardiovascular risk in patients with nephrotic syndrome. It provides new and strong evidence for the primary prevention of cardiovascular disease and the treatment of lipid metabolism disorders in patients with nephrotic syndrome.
METHODS In a single-center, retrospective nested case-control study, 350 patients with nephrotic syndrome at the First Affiliated Hospital of the Army Military Medical University between January 1, 1991, and November 30, 2019, were included in the study cohort. Of these, patients diagnosed with coronary artery disease (CAD) at the end of the observation period formed the case groups (n=115), or vice versa, the control groups (n=235). Obtain baseline information, duration of nephrotic syndrome, history of previous system diseases, HDL-C, LDL-C, eGFR, Alb, Fbg, and other biological test results. And also, statins using in treatment records as well as the accumulated medication duration. Use propensity score matching (ratio=1) to process the research cohort, construct contingency tables based on outcome events and statin usage, perform chi-square tests, and perform subgroup analysis based on different variables to draw forest maps. Perform binary logistic analysis to stratify by statins medication duration.
RESULTS Kept all the case group with propensity score matching (n=115), as well as the control group. Confounding of partly factors can be adjusted well. Chi-square test results shows that statin can reduce the risk of cardiovascular disease in patients with nephrotic syndrome (χ2=9.939, OR=0.355 (0.183–0.685), P=0.002). Subgroup analysis suggests that females, age over 60 years, no smoking, no alcohol consumption, course of NS over ten years, BMI>25 kg/m2, diabetes, eGFR≥90 and <60 mL/(min 1.73 m2), ALB<30 g/L, Fbg≤3.8 g/L, the effect on the outcome was not significantly different. Compared with patients without statin, treatment followed by between 2–6 months and 7–12 months is statistically significant (0.36 (0.18–0.71), P=0.003) and (0.24 (0.10–0.61), P=0.002) in binary logistic regression analysis, with no significant difference by more than 12 months (P=0.863).
CONCLUSIONS In patients with nephrotic syndrome who have dyslipidemia, statin therapy can decrease their CAD risk in the future. In a cumulative treatment duration of 2–12 months, with a risk reduction of CAD as the treatment term extension.
GW31-e0444
Chen Zhao1,2, Sining Hu1,2, Lulu Li1,2, Wei Meng1,2, Xi Chen1,2, Ming Zeng1,2, Bo Yu1,2, Haibo Jia1,2
1Department of Cardiology, The 2nd Affiliated Hospital of Harbin Medical University
2The Key Laboratory of Myocardial Ischemia, Chinese Ministry of Education, Harbin, China
OBJECTIVES Autopsy series showed that plaques underlying coronary thrombi in acute coronary syndromes (ACS) may have different features, and the most common phenotypes are plaque rupture and plaque erosion, which was affirmed by the optical coherence tomography (OCT) studies. Different plaque morphologies might be potentially amenable to different treatments, and identification of patients with of ACS resulting from erosion may permit a less invasive approach to management than the current standard of care. Recent studies have revealed that 30% of ST-segment elevation myocardial infarctions (STEMI) are caused by plaque erosion. Chronic inflammatory infiltration is a common process for atherosclerosis development. However, autopsy studies reveal that incidence rate of inflammatory infiltrates are less abundant in erosion compared with rupture. Studies performed by OCT have allowed to establish the severity of plaque inflammation by assessing macrophage infiltration. In this study, we aimed to assess the impact of macrophage infiltration on morphology of eroded plaque in patients with STEMI by OCT.
METHODS Consecutive patients with STEMI undergoing OCT imaging during primary percutaneous coronary intervention (PPCI) between October 2014 and December 2017 were included in this study. Among them, only patients with plaque erosion at the site of the culprit lesion were included for analysis. Plaque morphology were compared between the plaques with and those without macrophage.
RESULTS Three hundred and eight patients with plaque erosion were enrolled in this study finally. Macrophage infiltration was detected in 162 (52.6%) patients with plaque erosion at the site of culprit lesion, whereas 146 (47.4%) patients had no evidence of macrophage infiltration. Patients with macrophage infiltration were significantly younger (P=0.014) than those without macrophage. Patients presenting macrophage infiltration in the culprit plaques had a smaller minimal lumen area (MLA) (P<0.001) as compared with those without macrophage. Culprit segments exhibiting macrophage infiltration had thinner minimum fibrous cap thickness (FCT) (P<0.001). Of note, the maximal lipid arc and lipid core length were significant larger in culprit plaque with macrophage infiltration despite no difference was found in the latter. The subgroup analysis was performed to estimate the impaction of macrophage infiltration in culprit lesions progress by OCT from baseline to 1-year follow-up. The severity of culprit lesions was well decreased and stability of culprit plaques was well improved in both of two groups at 1-years follow-up. Moreover, the minimal FCT was significantly increased in patients with macrophage infiltration (P=0.046).
CONCLUSIONS Macrophage infiltration increased culprit lesion severity and plaque vulnerability in patients with STEMI caused by plaque erosion. These results may help to further understand the pathophysiology of plaque erosion and potentially tailored treatment strategies.
GW31-e0446
Xiaomin Tian, Heyu Meng, Fanbo Meng
China-Japan Union Hospital of Jilin University
OBJECTIVES AGPAT1 gene encodes 1-acyl-sn-glycerin-3-phosphate acyltransferase α, also known as LPA acyltransferase (LPAAT), is a key enzyme in the biosynthesis of phospholipids and triglycerides. It catalyzes the conversion of lysophosphatidic acid to phosphatidylcholine, and participates in cell signal transduction, inflammation, blood clot formation and other important biological processes. Inflammatory reaction and thrombosis are important pathological processes in the occurrence and development of coronary heart disease and myocardial infarction. The aim of this study was to evaluate whether the low expression of agpat1 gene in peripheral blood is associated with the risk of acute myocardial infarction (AMI), and whether it can be used as a biomarker to predict the risk of AMI.
METHODS Peripheral blood was collected from 91 patients with stable coronary heart disease and 95 patients with acute myocardial infarction. The mRNA expression of AGPAT1 gene was detected by real-time fluorescent quantitative polymerase chain reaction (PCR), and the expression level of AGPAT1 gene was detected by Western blot analysis at protein level.
RESULTS The results showed that the expression level of AGPAT1 gene in peripheral blood of AMI patients was significantly lower than that in stable CAD patients at RNA and protein levels (z=-2.358, P=0.018). There was no correlation between agpat1 gene expression and age, blood pressure, blood glucose, blood lipid level, smoking history and drinking history. The expression level of AGPAT1 gene in peripheral blood of patients with acute myocardial infarction is 0.28 times higher than that of patients with stable coronary heart disease.
CONCLUSIONS AGPAT1 gene expression in peripheral blood of patients with acute myocardial infarction was significantly lower than that in patients with stable coronary heart disease. Its low expression is an independent risk factor and related to the occurrence of acute myocardial infarction, which can be used as a biomarker to predict acute myocardial infarction.
GW31-e0448
Yifan Chen1,2, Genshan Ma1,2
1Cardiology Department of Nanjing Zhongda Hospital
2School of Medicine, Southeast University
OBJECTIVES This study was sought to compare of kissing balloon dilation (KBD) versus snuggling balloon dilation (SBD) on stent deformation in bifurcations.
METHODS Three stenting techniques of DK-Crush, DK-Culotte and T-stent were simulated on a bifurcation model, each stenting technique repeated 3 times, resulting in 18 stented phantoms, and then each phantom received either KBD or SBD, finally yielding 9 KBD and 9 SBD treatment. Micro-CT was performed on the stented phantoms to qualitatively grade stent deformation as mild, moderate and severe, to quantitatively measure the lumen over-expansion index (LOEI) and lumen asymmetric index (LASI) in the bifurcation core and its adjacent regions.
RESULTS The baseline features between KBD and SBD were similar in stent diameter and balloon diameter for main- and side-branch, KBD and SBD inflation pressure and distal bifurcation angle. Whatever stenting techniques being used, KBD produced severe stent deformation in and near the bifurcation core in all phantoms while SBD induced only mild stent deformation in one phantom in DK-Culotte, with severe stent deformation rate of 100% (9/9) for KBD vs. 0% (0/9) for SBD (P<0.001). Compared to KBD, LOEI or LASI was much lower in SBD (LOEI: 1.05±0.06 vs. 1.36±0.15, P<0.01; LASI: 1.03±0.04 vs. 1.26±0.10, P<0.01).
CONCLUSIONS SBD doesn’t cause and KBD frequently causes severe bifurcated stent deformation, indicating SBD is superior to KBD in optimization of bifurcated configuration in 2-stent techniques.
GW31-e0450
Yu Ning, Guihao Chen, Jingang Yang, Chunyan Tian, Yanyan Zhao, Xiaoyu Zhang, Yang Wang, Haiyan Xu, Yuejin Yang
State Key Laboratory of Cardiovascular Diseases, Fuwai Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College
OBJECTIVES Compared with America or Europe, the situation of cardiogenic shock (CS) complicating ST-elevation myocardial infarction (STEMI) in Asia is less known. The present study aimed to reveal the incidence rate, management, and in-hospital mortality of patients with CS complicating STEMI (STEMICS) in the whole China and at different-level (provincial, prefectural and county-level) hospitals.
METHODS We queried the 2013–2016 China Acute Myocardial Infarction (CAMI) registry databases to identify all patients with STEMI and/or CS (developing before or during hospitalization). We analyzed the incidence, management, and in-hospital mortality of patients with STEMICS in the whole China and at provincial, prefectural, and county-level and hospitals. Multivariable logistic regression models were used to identify predictors of CS and in-hospital death.
RESULTS The CAMI Registry enrolled 40,200 patients with acute myocardial infarction from 2013 to 2016. We finally included 28,054 STEMI patients in the study cohort. Of 2273 patients (8.1%) had CS, including 1132 (4.0%) with prehospital CS (developing before hospitalization) and 1141 (4.1%) with in-hospital CS (developing during hospitalization). Twenty-five thousand seven hundred eighty-one patients had no CS. 29.7% of the patients with overall STEMICS underwent primary percutaneous coronary intervention (PPCI). Rates of PPCI were 33.9 and 25.5% in patients with prehospital and in-hospital STEMICS respectively. In-hospital mortality were 49.8% in overall STEMICS, 33.7% in prehospital STEMICS, and 65.6% in in-hospital STEMICS. PPCI was an independent predictor of in-hospital STEMICS (odds ratio, 0.72; 95% CI, 0.59–0.88; P=0.0015) and in-hospital death in patients with prehospital STEMICS (odds ratio, 0.53; 95% CI, 0.33–0.83; P=0.0062). In terms of hospital level, the incidence rates of STEMICS (overall, prehospital, and in-hospital STEMICS) and in-hospital mortality of patients with prehospital STEMICS from county-level hospitals were dramatically higher than those from provincial or prefectural hospitals, which might be associated with the much lower PPCI rates in the patients from county-level hospitals.
CONCLUSIONS Rates of STEMICS in China between 2013 and 2016 were consistent with those in western countries. However, the PPCI rates of STEMICS in China were relatively low and in-hospital mortality were dramatically high. In China, STEMICS patients from county-level hospitals had higher rates of CS and in-hospital mortality but lower PPCI rate than those from provincial or prefectural hospitals. PPCI was an independent protective factor against CS developing during hospitalization and in-hospital death in patients with prehospital STEMICS.
GW31-e0451
Xunxun Feng, Qianyun Guo, Guangyao Zhai, Yujie Zhou
Capital Medical University, Beijing Anzhen Hospital
OBJECTIVES Patients that undergo renal transplantation (RT) often suffer from high rates of cardiovascular disease-related mortality, yet there have not been sufficient angiographic studies of coronary artery disease (CAD) findings in RT patients conducted to date.
METHODS This study examined coronary angiography findings from 45 patients with functional renal grafts for over 6 months that were analyzed in Anzhen Hospital (Beijing, China) from 2014 to 2019. For comparison purposes, we additionally examined coronary angiography findings from 45 age- and sex-matched patients undergoing chronic dialysis due to end-stage renal disease (ESRD). We used the SYNTAX score to gauge CAD severity.
RESULTS The duration of ESRD in patients in the RT group was significantly longer than for that of patients in the dialysis comparison group (19.31±7.83 vs. 11.43±8.04 years, P<0.001). The SYNTAX scores for patients in the dialysis and RT groups were 17.76±7.35 and 12.57±5.61, respectively (P<0.01). We found that 64.4 and 28.9% of dialysis and RT patients, respectively, exhibited the presence of moderate or severe calcified lesions upon examination. In addition, the SYNTAX scores of RT patients were correlated with ESRD duration (P<0.001).
CONCLUSIONS We observed less serious CAD in RT patients relative to long-term dialysis patients even though the former group exhibited a longer mean ESRD duration. Both groups exhibited high rates of calcification of the coronary artery, even following RT.
GW31-e0453
Xianjing Wei, Lianna Xie
The Affiliated Zhongshan Hospital of Dalian University, Dalian
OBJECTIVES Although the right radial artery access is considered as the default technique for Percutaneous Coronary Intervention access, the incidence of radial artery occlusion is still relatively high, discomfort from perioperative patients is common and Inconveniences are overflowed for operators whose patient’s right radial occlusion, underdeveloped right radial artery, extreme right radial tortuosity, sclerosis or calcifications, arteria lusoria, previous right radial failure, presence of an arteriovenous shunt in the right arm, post-CABG patients requiring LIMA angiography. The aim of this report is to evaluate the feasibility of the left distal transradial (lDTRA namely, anatomical snuffbox) assess.
METHODS Of 550 consecutive patients needed coronary angiography or intervention therapy in the Affiliated Zhongshan Hospital of Dalian University, from January 2019 to January 2020, no forearm artery ultrasound before operation, were stratified into two groups: group A: the patients underwent right radial artery puncture; group B: underwent LDTRA puncture. We analyze the safety and Feasibility between 2 groups. Feasibility was the success rate of cannulate the distal left radial artery and completing Coronary Intervention detail: X-ray exposure time, the number of angiographic catheters to per patient, the number of completed target revascularization, complexity of coronary intervention. The safety point included hematoma, bleeding or neuropathy.
RESULTS Of 260 consecutive patients assigned to group A, 248 cases underwent (Success rate of 95.38%) right radial puncture, 106 cases were cannulated 6F radial artery sheaths and 142 cases were cannulated 5F radial artery sheaths; Group B was attempted in 290 patients, 277 cases (95.52%) had successful completion of the procedure, 96 cases were cannulated 6F radial artery sheaths and 181 cases were cannulated 5F radial artery sheath. The success rate between the two group is no statistical significance, P<0.05. There was no significant difference between the two groups in the details of coronary intervention (the number of angiographic catheters to per patient, the number of completed target revascularization, complexity of coronary intervention) P<0.05. However, the rate of forearm hemorrhage and radial artery occlusion during hospital to LDTRA is lower than that of right radial artery approach.
CONCLUSIONS LDTRA is a safe and feasible arterial access. LDTRA provides improved operator ergonomics and patient’s comfort.
GW31-e0455
Xunxun Feng, Qianyun Guo, Guangyao Zhai, Yujie Zhou
Capital Medical University, Beijing Anzhen Hospital
OBJECTIVES Serum uric acid (SUA) levels has been considered a possible risk factor for coronary artery disease (CAD) for many years. Since SUA levels is greatly affected by the body’s medication therapies, dietetic status, and metabolism, the interactions between SUA and CAD have been controversial for centuries. However, the state of hyperuricemia (HUA) has been proven to have a negative impact on CAD in previous studies, but there is still few clinical and epidemiological evidence of HUA in CAD. In view of the fact that there are few studies on postmenopausal women, this study explored the influence of SUA levels and HUA on CAD in this demographic group.
METHODS In total, 5435 postmenopausal women were allocated to either a non-CAD group (n=2021) or a CAD group (n=3414). Regression analysis, correlation analysis, comparison between stratified groups, and statistical analysis of diuretics were carried out on data obtained in this study.
RESULTS In the multivariate analysis, we found that SUA and HUA were independent risk factors for CAD (SUA OR model 1 to model 4: 1.004, 1.004, 1.004 and 1.003; HUA OR model 1 to model 4: 1.844, 1.837, 1.731 and 1.486). Correlation analysis showed that SUA and HUA positively correlated with CAD (P<0.001). By comparing the stratified age groups, we found that the differences between the groups were significant (P<0.05).
CONCLUSIONS SUA levels might be greatly affected by age while HUA could be influenced by diuretic use, so the effects of both age and diuretic use on SUA levels and HUA should not be ignored. To some extent, SUA and HUA could be independent risk factors for postmenopausal women with CAD.
GW31-e0457
Zhiqing Qiao, Xuedong Shen, Ying Zheng, Wei Wang, Hang Zhao, Jun Pu
Renji Hospital, Shanghai Jiao Tong University School of Medicine
OBJECTIVES Myocardial work echocardiography (MWE) is a new method to evaluate myocardial contraction, which is based on the left ventricular pressure and speckle tracking echocardiography. This study was aimed to analyze the relationship between the measurements of MWE and outcome in patients with acute ST-elevated myocardial infarction (STEMI) after primary percutaneous cardiac intervention (PCI) during hospitalization.
METHODS The study included 42 patients with acute STEMI who successfully underwent primary PCI (37 men, age 61.9±9.5 years). The 42 patients underwent MWE within 24 hours after PCI, and 25 of them underwent MWE again within 2–9 days after PCI (mean 4.36±2.02 days). The measurements included global myocardial work index (GWI), global myocardial constructive work (GCW), global myocardial wasted work (GWW), global myocardial work efficiency (GWE). Each segment of left ventricle was further analyzed for pressure-strain loop to evaluate its shape, orientation and size. Other parameters included brain natriuretic peptide (BNP) and left ventricular ejection fraction (LVEF).
RESULTS Five patients had major adverse cardiac events (MACE) after PCI, including 1 death and 4 acute heart failures during hospitalization. There were significant improved in LVEF, GWI, GCW, GWW and GWE (P=0.002–0.04) at second follow up after PCI, but not in BNP (P=0.06). The sensitivity and specificity of GWE predicting MACE were 83 and 81%, respectively (cutoff point<75.7%, AUC=0.86, P=0.0001). Multivariate regression analysis showed that GWE independently predicted MACE (OR=0.89, 95% CI=0.81–0.98, P=0.02). Further analysis showed that the shape, orientation and size of left ventricular pressure-strain loop in ischemic segment had no significantly improved in 8 of 25 patients at follow up. The shape of the loop was narrow and it shifted to right in those patients. The incidence of MACE in patients who had unimproved loops in ischemic segments was significantly higher than that in patients who had improved loops (50.0 vs. 5.9%, P=0.01).
CONCLUSIONS MWE provides us a new sight to evaluate the outcome after PCI. It is of vital importance to explore the regression of ischemic segments after PCI.
GW31-e0463
Thach Nguyen1,2, Nguyen Thanh Luan2, Vy Le2, Duy Chung1, Tra Ngo1, Luan Ngo1, Phuong M. Nguyen1, Phuoc T. Nguyen1, Thai Truong1, An T. Ngo1, Hoang C. Nguyen1, Quang N.N. Do2, Hien D.N. Duong2, Vu Tri Loc2, Tran P.H. Nhan3, Cao Van Thinh4, Ho Thuong Dung5, Gianluca Rigatelli6, Aravinda Nanjundappa7
1Methodist Hospital, Merrillville IN
2Tan Tao University School of Medicine
3Tam Duc Hospital, Hochiminh City, VN
4Pham Ngoc Thach School of Medicine, Hochiminh City, VN
5Thong Nhat Hospital, Hochiminh City, VN
6Cardiovascular Diagnosis and Endoluminal Interventions, Section of Adult Congenital Interventions, Rovigo General Hospital, Rovigo, Italy
7The University of West Virginia Medical Center, Charleston, WV
OBJECTIVES Coronary injuries are hypothesized to be caused by the cavitation phenomenon (explosion of air bubbles) which is seen frequently in domestic or industrial pipes. Following hydraulics principle, with distal negative suctioning in diastole, if the coronary dynamic pressure decreases below the vapor pressure (VP) most likely of nitrogen in the blood, bubbles would form. They explode when the coronary dynamic pressure recovers>the VP during systole. These explosions create jet waves weakening and rupturing the cap of the plaque, triggering acute coronary syndrome (ACS). How could these events be located, recorded and tabulated?
METHODS Angiograms with ACS culprit lesions were selected. The left coronary arteries were recorded in the right anterior oblique caudal view and the right coronary artery in the left anterior oblique view (at 15 frames per second). Then the angiograms were viewed off line frame by frame. The first frame was the angiogram of an artery completely filled with contrast. The following frames showed the blood moving in, seen in white. The flow could be LAMINAR (Figure 1A and B). TURBULENT (mixing of blood in white and contrast in black) (Figure 2) or RETROGRADE (black column traveling backward). The turbulent flow reflects the collision between antegrade and retrograde flow. The LOCATION and the length in TIME of laminar, retrograde and mainly turbulent flow were recorded. The intensity of turbulent flow was measured by (1) the length of coronary segment with mixing contrast and blood (2) the length of the stagnant retrograde flow.
RESULTS The results of 50 angiograms with ACS showed that after being laminar (85%) at the beginning of diastole, the flow became turbulent with diffuse mixing of black (contrast) and white (blood) at the MID SEGMENT of the LAD, LCX or RCA. This observation matched with the location of 82% of ruptured plaques. The length of the time of retrograde flow lasted more than 30 frames encompassing 2 systoles.
CONCLUSIONS This is the first time, the matching of location of ruptured plaques and turbulent flow representing the collision between antegrade flow in diastole and retrograde flow in systole was confirmed. These results may help to find the precise measures preventing ACS.
GW31-e0490
Xiangbin Zhong, Yifen Lin, Shaozhao Zhang, Xiaodong Zhuang, Xinxue Liao
First Affiliated Hospital of Sun Yat-Sen University
OBJECTIVES Cardiovascular disease remains the leading cause of mortality world wild. Coronary heart disease (CHD) accounts for the greatest proportion of CVDs. Coronary heart disease was thought to be an infirmity of age, but the proportion of AMI hospitalizations attributable to young patients increased and was most pronounced among women, which indicates the risk factors has already accumulated among sex even in the youth. Risk assessment with long-term prediction algorithms can help to identify individuals who would benefit most from risk-factor interventions [1], but failed in suggesting the difference proportion among sex in youth. To further optimize preventive strategies for coronary heart disease among sex, it is essential to understand and appropriately quantify the mediation differences of gender on CHD by traditional cardiovascular factors [2].
METHODS This investigation included Framingham Heart Study offspring cohort participants attending their first examination cycle (1971–1975), when traditional risk factors data were first reliably collected. During the following, newly onset coronary heart disease before 2010 was recorded. Written informed consent was obtained from participants for all study procedures. Mediation analysis was used to compare the mediation proportion of sex on Coronary Heart Disease by traditional cardiovascular factors mentioned in Framingham risk score. Each study was approved by relevant local or national ethics committees and all procedures performed in these studies were in accordance with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Statistical analysis was performed by R version 3.5.2, while mediation analysis was achieved by package “Mediation”.
RESULTS Of 5013 offspring cohort participants, consists of 2427 males and 2586 females were enrolled. 615 male (25.3%) and 287 (11.1%) female adjudicated to have a CHD during the study period. All of the traditional risk factors showed statistically different distribution between males and females (Table 1).
CONCLUSIONS The different mediation of sex on CHD was mostly contributed by blood pressure (DBP 31% or SBP 26%). High-density cholesterol and low-density cholesterol (or total cholesterol) provided 23 and 19% (or 13%) difference. Age, smoking status, and diabetes provided less than 10% contribution to CHD risk. The imbalanced distribution of traditional risk factors made up to 88% differences in CHD risk among males and females.
GW31-e0496
Chendie Yang, Xiaoqun Wang
Department of Cardiology, Ruijin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, P.R. China
OBJECTIVES Patients with type 2 diabetes are under substantially higher risk of in-stent restenosis (ISR) after coronary stent implantation. We sought to investigate whether visit-to-visit glycemic variability is a potential predictor of ISR in diabetic patients after stent implantation.
METHODS Type 2 diabetic patients underwent elective percutaneous coronary intervention were consecutively enrolled and 1-year follow-up coronary angiography was performed. The incidence of ISR and its relationship with visit-to-visit HbA1c variability, expressed as coefficient of variation (CV), standard deviation (SD) and variability independent of the mean (VIM), were studied. Multivariable Cox proportional hazards models were constructed to analyze the predictive value of glycemic variability for ISR.
RESULTS From September 2014 to July 2018 in Ruijin Hospital, a total of 420 diabetic patients (688 lesions) after stent implantation were included in the final analysis. During a mean follow-up of 12.8±1.3 months, the incidence of ISR was 8.6%, which was significantly increased in patients with higher CV of HbA1c (P=0.001). The mean diameter stenosis (DS), net luminal loss and net luminal gain were 22.9±16.8%, 0.42±0.88 mm and 1.66±0.83 mm, respectively. Greater DS was observed in subjects with higher tertiles of CV of HbA1c (P<0.001), and this trend was more prominent in patients with optimal glycemic control (HbA1c≤7%). In multivariate analysis, HbA1c variability was independently associated with incidence of ISR after adjustment for traditional risk factors and mean HbA1c (HR: 3.00 [95% CI: 1.14~7.92] for highest vs. lowest tertile). Inclusion of CV of HbA1c led to a better risk stratification accuracy. Assessing glycemic variability by SD or VIM yielded similar findings.
CONCLUSIONS This study suggests that visit-to-visit HbA1c variability is an independent predictor of incidence of ISR in patients with type 2 diabetes after stent implantation.
GW31-e0516
Chendie Yang, Xiaoqun Wang
Rui Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, P.R. China
OBJECTIVES In-stent restenosis (ISR) remains a major limitation of percutaneous coronary intervention despite improvements in stent design and pharmacological agents, whereas the mechanism of ISR has not been fully clarified. In the present study, we sought to investigate the potential association of serum soluble TREM-1 (sTREM-1) levels with the incidence of ISR. The role of TREM-1 was evaluated in cultured vascular smooth muscle cells (VSMCs).
METHODS In 1683 patients undergoing coronary intervention and follow-up coronary angiography after approximately one year, 130 patients were diagnosed with ISR, and 150 gender- and age-matched patients with no ISR were randomly included as controls. Levels of sTREM-1 were determined by ELISA. The role of TREM-1 signaling in the activation of VSMCs was tested.
RESULTS Serum sTREM-1 concentrations were significantly elevated in patients with than without ISR. Multivariable logistic regression analysis showed that sTREM-1, besides conventional factors, was independently associated with the incidence of ISR. Evident expression of TREM-1 in VSMCs was detected both in the neointimal and medial layers of stenotic lesions of mouse carotid ligation models. In cultured VSMCs, expression of TREM-1 was significantly induced upon exposure to lipopolysaccharide. Blocking of TREM-1 with a synthetic inhibitory peptide LP17 dramatically inhibited, whereas TREM-1-activating antibody promoted cellular inflammation, proliferation and migration in VSMCs.
CONCLUSIONS These data suggest that TREM-1 is a predictive biomarker of ISR and an important mediator of cellular inflammation, migration, and proliferation in VSMCs. Pharmacological inhibition of TREM-1 may serve as a promising approach to attenuate the progression of ISR.
GW31-e0519
Xiaoqun Wang
Rui Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, P.R. China
OBJECTIVES High-density lipoprotein (HDL) is a pluripotent atheroprotective factor with reverse cholesterol transport, anti-inflammatory, and pro-angiogenic activities. Increasing evidence suggests that HDL function rather than concentration would be a more relevant measure to predict cardiovascular risk. In this study, we sought to investigate whether cholesterol efflux capacity (CEC) of HDL is related to coronary collateralization in patients with stable angina and chronic total occlusion.
METHODS CEC was determined in vitro in 115 patients with stable angina and angiographic total occlusion of at least one major coronary artery. The degree of coronary collaterals supplying the distal aspect of a total occlusion from the contralateral vessel was graded according to Rentrop classification.
RESULTS CEC of HDL was significantly elevated in good collateralization (Rentrop score of 2–3) than poor collateralization (Rentrop score of 0–1) group (15.39±10.17% vs. 10.18±5.41%, P=0.004). While CEC was positively correlated to Rentrop score (Spearman’s r=0.275, P=0.005), no correlation was observed between Rentrop score and serum levels of HDL-cholesterol or apolipoprotein A. Receiver operating characteristic curve yielded an AUC of 0.662 (95% confidence interval: 0.558–0.767, P=0.006) in discriminating good or poor collateralization. After adjusting for age, gender, body mass index and conventional risk factors for coronary artery disease in multivariate logistic regression analysis, CEC persisted to be an independent determinant for good collateralization (odds ratio: 1.11 per 1% increase, P=0.006).
CONCLUSIONS Increased CEC of HDL is associated with good coronary collateralization in patients with stable angina and chronic total occlusion.
GW31-e0522
Xiaojiao Zhang, Aijie Hou, Bo Luan
Department of Cardiology, The People’s Hospital of China Medical University, The People’s Hospital of Liaoning Province
OBJECTIVES To assess the effectiveness and safety of ARW for vascular recanalization in CTO patients.
METHODS A total of 301 consecutive CTO patients who received the antegrade percutaneous coronary intervention (PCI) between December 2015 and December 2019 at our institution were included, of whom 11 were treated with ARW (10 successfully) for vascular recanalization. The applicability and safety of ARW were assessed.
RESULTS Among the 301 CTO patients who received antegrade vascular recanalization, 11 were treated with ARW. The following treatment approach was successful in 10 patients as follows: from the diagonal branch (D) to anterior descending branch (LAD) in 4 patients; from the septal branch (S) to LAD in 1 patient; from D to S and LAD in 1 patient; from the circumflex branch (LCX) to obtuse marginal branch (OM) in 1 patient; from OM to LCX in 1 patient; from a posterior descending artery (PDA) to the posterior lateral vein (PLV) in 2 patients. Yet, the treatment in the other patient with RCAm CTO failed, while the consequent reverse vascular recanalization succeeded. The mean J-CTO score of the 11 patients was 2.7±0.65, among whom eight were accompanied with calcifications. Sion Black and Fielder XT-R reverse wires were used in 9 and 2 patients, respectively. No loss of side branches or severe surgery-related complications occurred in 11 patients.
CONCLUSIONS Therefore, WRW can improve surgical efficiency and should be popularized for further application.
GW31-e0588
Yuhong Zeng, Dong Zhao
Department of Epidemiology, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart, Lung and Blood Vessel Diseases
OBJECTIVES Recently, Chinese Society of Cardiology (CSC) expert consensus on lipid management of extreme high-risk atherosclerotic cardiovascular disease (ASCVD) patients recommended further risk stratification among patients with ASCVD. A new definition of category of “extreme high-risk” has been proposed by this expert consensus, along with, the new and lower treatment target of low-density lipoprotein cholesterol (LDL-C) was recommended for the patients. This study aimed to assess the expanding needs on lipid-lowering treatment by applying the new risk category among patients with acute coronary syndrome (ACS).
METHODS The study was based on the Improving Care for Cardiovascular Disease in China (CCC) project, which was launched in 2014 as a collaborative initiative of the American Heart Association and the CSC. This study enrolled ACS inpatients in CCC from 240 hospitals nationwide from November 2014 to July 2019. The proportion of patients at extreme high-risk, mean LDL-C levels at admission and the median distance between LDL-C level and the new target were assessed.
RESULTS Among 104,516 ACS inpatients enrolled in this study, 75.1% (78,521) met the criteria of extreme high-risk and were expected to achieve the new LDL-C goal. Among patients at extreme high-risk, 21.2% (16,651) had multiple severe ASCVD events and 78.8% (61,870) had 1 severe ASCVD event and at least two high-risk factors. For the extreme high-risk patients, the mean level of LDL-C was 106.5±39.3 mg/dL, 92.8% of them had LDL-C=55 mg/dL at admission and the median distance between LDL-C level and the target of 55 mg/dL was 51.3 (30.1, 76.1) mg/dL.
CONCLUSIONS About three fourth of inpatients with ACS were categorized as extreme high-risk based on the definition of CSC expert consensuses, nine out of ten patients at extreme high-risk didn’t achieve the new LDL-C target at admission. There are substantially expanding needs for more effective lipid-lowering strategies.
GW31-e0596
Qian Zhang, Daoyuan Si, Zhongfan Zhang, Haikuo Zheng, Wenqi Zhang
China-Japan Union Hospital of Jilin University
OBJECTIVES The predictors of left ventricular thrombus (LVT) formation are not well-defined in the contemporary era, especially in those patients at high risk. We aimed to evaluate whether platelet-to-lymphocyte ratio (PLR) be valuable in determining LVT formation in patients with anterior ST-Elevation Myocardial Infarction (STEMI) and left ventricular (LV) dysfunction.
METHODS LVT group (n=46) were identified from the anterior STEMI patients with LV dysfunction, who were treated with the primary percutaneous coronary intervention (PCI) from January 2017 to December 2019 in China-Japan Union Hospital of Jilin University. The controls (n=92) were also selected from the same batch patients, with age and gender matched and without LVT. PLR were determined at admission, which was calculated as the ratio of the platelet count to the lymphocyte count using complete blood count. LVT was defined by echocardiography.
RESULTS The PLR levels were significantly higher in patients with LVT than those in controls (P=0.001). In a receiver operator characteristic curve (ROC) analysis, using a cut-off value of 118.07 (AUC 0.673, 95% CI: 0.574–0.771, P=0.001) could independently predict the occurrence of LVT. Multivariate analysis showed that increased PLR (OR=1.011, 95% CI: 1.004–1.018, P=0.002), left ventricular aneurysm (OR=46.350, 95% CI: 5.659–379.615, P<0.001) and DTBT (OR=1.005, 95% CI: 1.001–1.009, P=0.012) were independent predictors in predicting LVT formation.
CONCLUSIONS In acute anterior STEMI patients with LV dysfunction, PLR is an independent predictor of LVT formation. A larger prospective study is warranted to evaluate this result.
GW31-e0598
Na Yang1, Jing Liu1, Jun Liu1, Yongchen Hao1, Yuhong Zeng1, Sidney C. Smith Jr2, Yong Huo3, Gregg C. Fonarow4, Junbo Ge5, Kathryn A. Taubert6, Louise Morgan7, Changsheng Ma8, Yaling Han9, Dong Zhao1
1Department of Epidemiology, Beijing An Zhen Hospital, Capital Medical University, Beijing Institute of Heart, Lung and Blood Vessel Diseases, Beijing, China
2Division of Cardiology, University of North Carolina, Chapel Hill, NC
3Department of Cardiology, Peking University First Hospital, Beijing, China
4Division of Cardiology, Geffen School of Medicine at University of California, Los Angeles, CA
5Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, China
6Department of International Science, American Heart Association, Basel, Switzerland
7International Quality Improvement Department, American Heart Association, Dallas, TX
8Department of Cardiology, Beijing An Zhen Hospital, Capital Medical University, Beijing, China
9Cardiovascular Research Institute and Department of Cardiology, General Hospital of Shenyang Military Region, Shenyang, Liaoning, China
OBJECTIVES To evaluate the effect of ticagrelor plus aspirin in in-hospital outcomes compared with clopidogrel plus aspirin in patients 75 years and older with acute coronary syndrome (ACS) in a real-world population.
METHODS The Improving Care for Cardiovascular Disease in China-ACS project is an ongoing quality improvement project of the American Heart Association and Chinese Society of Cardiology. The primary effectiveness outcome was in-hospital major adverse cardiovascular event (MACE) including cardiac death, myocardial infarction (MI), stent thrombosis, and ischemic stroke during hospitalization. The major safety outcome was major bleeding. Cox proportional hazard models adjusting for confounders were used to evaluate the effect of ticagrelor plus aspirin in in-hospital outcomes compared with clopidogrel plus aspirin. A propensity score-matched analysis was also conducted.
RESULTS From November 2014 to December 2019, 18,244 ACS patients aged 75 years and older receiving dual antiplatelet therapy (3566 received ticagrelor plus aspirin and 14,678 received clopidogrel plus aspirin) were included. Patients receiving ticagrelor plus aspirin were more likely to be ST-segment-elevation myocardial infarction and male, and had less comorbidities. The MACE had occurred in 4.57% of the patients receiving ticagrelor plus aspirin as compared with 4.28% of those receiving clopidogrel plus aspirin (hazard ratio (HR): 1.14; 95% confidence interval (95% CI): 0.93–1.39; P=0.20). The major bleeding had occurred in 3.31% of the patients receiving ticagrelor plus aspirin as compared with 2.64% of those receiving clopidogrel plus aspirin (HR: 1.00; 95% CI: 0.74–1.35; P=0.99). Among 6396 propensity score-matched patients, 4.47% of the patients receiving ticagrelor plus aspirin had MACE, while 4.00% of the patients receiving clopidogrel plus aspirin had MACE (HR: 1.18; 95% CI: 0.90–1.55; P=0.24). A total of 3.00% of the patients receiving ticagrelor plus aspirin had major bleeding, while 2.41% of the patients receiving clopidogrel plus aspirin had major bleeding (HR: 1.08; 95% CI: 0.69–1.69; P=0.73).
CONCLUSIONS In patients with ACS and aged 75 years and older, ticagrelor had comparable effects with clopidogrel in reducing the occurrence of in-hospital MACE and major bleeding.
GW31-e0601
Yong Liu1, Yibo He1, Liwei Liu1, Guoli Sun1, Shiqun Chen1, Jin Liu1, Wenhua Liang2, Chunquan Ou3, Kaihong Chen4, Liling Chen4, Jianfeng Ye5, Yan Liang6, Yunzhao Hu7, Jie Li1, Xin Li1, Ning Tan1, Jiyan Chen1
1Department of Cardiology, Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of Coronary Disease, Guangdong Provincial People’s Hospital, South China University of Technology, Guangzhou 510100, China
2Department of Pneumology, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China
3Department of Statistics, Southern Medical University, Guangzhou, China
4Longyan First Affiliated Hospital of Fujian Medical University, Longyan, China
5Dongguan TCM Hospital, Dongguan, China
6Maoming People’s Hospital, Maoming, China
7Shunde Hospital of Southern Medical University, Shunde, China
OBJECTIVES To assess the tendency of acute myocardial infarction (AMI) patients admission during and after the COVID-19 outbreak.
METHODS This study is a cross-sectional study. Admission data of acute myocardial infarction patients were enrolled in five PCI centers, which were also the designated hospital for COVID-19 patients of the 5 cities in southern China. The section of pandemic period of COVID-19 was defined as January 23, 2020 to February 24, 2020, when public health First Level Emergency Response were carried out by provincial health commission. AMI admission of pandemic period were compared with pre-pandemic period (January 1–22, 2020), recovery period following pandemic (February 25 till April 30, 2020). Considering the influence of China Spring Festival, the adjusted section of the former corresponding period of pandemic period in 2019 was defined as January 12 till May 10, 2019. The cross-sectional number of AMI patients admission were compared to assess the difference with the use of Poisson regression. Weekly admission was drawn to understand the tendency of how it changed along the pandemic period to the recovery period.
RESULTS During the COVID-19 pandemic period, the average admission rate of AMI patients was 5.45 per day, which was significant lower than that of pre-pandemic period and the former corresponding period in 2019 (8.32 and 10.38 patients per day respectively, P<0.001). As the pandemic period ended up, the AMI patients admission rate was 6.89 per day, significantly higher than the pandemic period, however, lower than the pre-pandemic period and the former corresponding period. Weekly admissions were drawn from January 1 to April 28, the number of AMI admission was on the rise, returning to the same level of the corresponding period in 2019 at the eighth week following the end of pandemic period.
CONCLUSIONS When the COVID-19 pandemic period end up, the number of AMI patients admission which have fallen since the outbreak would rebound gradually.
GW31-e0604
Pan Li, Wendong Tang, Xiaxian Shen, Pan Hou, Hailing Li, Xian Guo, Xianxian Zhao
Changhai Hospital, Shanghai
OBJECTIVES Early detection and prevention of high-risk coronary artery disease (CAD), especially in atypical patients, is clinically crucial to reduce cardiovascular death. Computed tomography angiography (CTA) or coronary angiography (CAG) have limited application as screening tools, due to invasiveness, costliness, radiation risk and potential renal injury. Carotid ultrasound is a noninvasive and sensitive screening method for detecting subclinical atherosclerosis and identifying CAD. The commonly used prediction index, the intima-media thickness (IMT) and carotid plaque score (PS) have been shown to be associated with CAD. However, recent evidence has proven that IMT has a weak predictive value of CAD. Moreover, the variation range in plaque thickness is small (approximately 0.01–0.1 mm), and the measurement process of PS is complicated and time-consuming; hence, following individual changes in atherosclerosis progression is difficult. In contrast, plaque length grows faster than its thickness and has a large dynamic range, which makes it convenient for clinical observation. Nevertheless, the relationship between ultrasound-based carotid plaque length (CPL) and CAD has not been fully explored. This study therefore compares CPL with coronary findings in patients with suspected CAD undergoing CAG.
METHODS We prospectively enrolled 2149 consecutive patients who underwent both first coronary angiography and carotid ultrasonography with measurements of IMT, PS and CPL (Figure 1).
RESULTS We prospectively enrolled 2149 consecutive patients who underwent both first coronary angiography and carotid ultrasonography with measurements of IMT, PS and CPL (Figure 1). In total, 1408 (65.5%) patients had CAD (defined as stenosis ≥50%), and 741 (34.5%) patients had no CAD. Patients with CAD had longer maximal CPL than those without CAD (P<0.001). The severity of CAD, measured by the Gensini score (GS), was closely correlated with max-CPL (r s=0.560), followed by PS (r s=0.486) and mean-IMT (r s=0.292) (Figure 2). Multivariate analysis revealed that max-CPL remained independently associated with CAD and high-GS after adjustment for traditional risk factors (TRF). Max-CPL, compared with PS or mean-IMT, had significantly higher discrimination value for predicting high-GS (area under the curve [AUC] 0.819 vs. 0.769 vs. 0.634, P<0.001) (Figure 3). At a cut-off value for the max-CPL of 6.3 mm, the sensitivity and negative predictive value for high-GS were 84.6 and 89.1%, respectively. Furthermore, the addition of max-CPL significantly improved the discrimination (AUC 0.832 vs. 0.720, P<0.001) and reclassification (net reclassification improvement [NRI]=0.431, P<0.001) over TRF for high-GS.
CONCLUSIONS Our study demonstrates that ultrasound max-CPL showed better independent and incremental value in predicting the presence and severity of CAD than PS or mean-IMT, thus suggesting that it may help identify high-risk CAD patients for widening the window for earlier prevention (Figure 4). However, further studies are required to confirm our findings and establish the link between max-CPL and prognosis of patients with CAD.
GW31-e0610
Zhicheng Hu, Yan Yao
Department of Cardiology, Fuwai Hospital, National Center for Cardiovascular Diseases, CAMS & PUMC
OBJECTIVES To investigate knowledge for non-vitamin K antagonist oral anticoagulants (NOACs) in Chinese doctors and identify the target doctors for continuing medical education.
METHODS Clinical doctors attending 2019 the 30th Great Wall International Congress of Cardiology were interviewed with modified Anticoagulation Knowledge Tool (AKT) including 10 questions regarding demographic data, clinical practice characteristics and 20 questions about knowledge of NOACs. Correction rate of knowledge questions over 80% is regarded as well-known for NOACs.
RESULTS A total of 578 valid questionnaires were collected from 597 ones. Among all investigated doctors, 39.4% were from north China. Males accounted for 50.9%. The average number of correctly-answered questions about knowledge of NOACs was 14.8±1.9. The overall well-known rate was 39.1%. The knowledge poorly-handled was how to treat overdose use (10.0%), the hazard of one does missing (31.1%) and how to reduce adverse side effects when taking NOACs (38.1%). The multivariate logistic analysis revealed that doctor degree acquired [odds ratio (OR)=3.70, 95% Confidential interval (CI)=1.49–9.17, P<0.01], practiced in Class A tertiary hospital (OR=1.51, 95% CI=1.07–2.13, P<0.05) predicted doctors had good knowledge of NOACs and worked in hospital in west China (OR=0.42, 95% CI=0.27–0.66, P<0.01) predicted doctors had poor knowledge.
CONCLUSIONS The majority of Chinese doctors had poor knowledge of NOACs, particularly in how to treat overdose use, the understanding of the hazard of one does missing and how to reduce adverse side effects of NOACs. Continuing medical education should be performed intensively to improve the quality of knowledge, especially for doctors who worked in non-Class A tertiary hospital and west China.
GW31-e0612
Haowei Li, Yue Qi
Department of Epidemiology, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart, Lung and Blood Vessel Diseases, Beijing 100029, China
OBJECTIVES Blood concentrations of triglycerides (TG) are generally considered to reflect atherogenic lipoproteins. Treatment strategies to improve TG clearance or reduce TG production have become a central topic in the debate over residual risk of ASCVD. Guidelines have recommended patients with high TG and high cardiovascular risk or ASCVD should lowering TG. However, there are significant differences in the recommendations of guidelines for the management of patients with high TG levels, especially in those with acute coronary syndrome (ACS), considering their extremely high risk. It remains unclear whether triglyceride lowering is effective in preventing ASCVD in patients with residual risk. This study aims to evaluate whether triglyceride level is associated with the risks of in-hospital major adverse cardiovascular and cerebrovascular events (MACCE) for ACS inpatients in China.
METHODS The Improving Care for Cardiovascular Disease in China-ACS Project is ongoing nationwide registry of the American Heart Association and the Chinese Society of Cardiology. A total of 104,513 inpatients with a definitive diagnosis of ACS were included. TG levels measured at admission were divided into lower (<200 mg/dL) and higher (≥200 mg/dL) groups. Multivariate logistic regression was performed to examine the association between triglyceride level and in-hospital MACCE outcomes, and a propensity-score-matched analysis was further conducted.
RESULTS Among these ACS patients, 23,685 (22.6%) patients had TG ≥200 mg/dL, and even 39.0% patients had TG ≥150 mg/dL. Patients with high TG were younger, with high proportion of cardiovascular risk factors such as smoking, hypertension and diabetes, but had lower comorbidities and less severe clinical conditions. Multivariate logistic analysis identified TG level as a negative predictor for in-hospital MACCE. TG ≥200 mg/dL was significantly associated with 27% lowering risk of MACCE (OR 0.73, 95% CI 0.62–0.82, P<0.001). The results were similar among STEMI and NSTE-ACS patients and even after propensity score-matched analysis.
CONCLUSIONS High level of triglyceride was highly prevalent in ACS inpatients in China. On-admission TG levels was inversely related with MACCE. TG level may be regarded as an indicator of good nutritional status that protects such patients from short cardiovascular events rather than a hazard factor, and therefore does not warrant aggressive treatment to lowering it to the so-called normal level.
GW31-e0630
Haixia Qin, Zhenbing Liu
Ordos Central Hospital
OBJECTIVES To evaluate the protective effect of Shexiang Baoxin Pill (SBP) on MI/RI and myocardial infarction area in STEMI patients.
METHODS One hundred and three STEMI patients were randomly divided into PPCI group (patients only receiving PPCI) (n=52) and PPCI+SBP group (patients receiving SBP and PPCI) (n=51). The area at risk of infarction (AAR) and final infarct size (FIS) were examined by gated single photon emission CT (SPECT). MI/RI was assessed using myocardial salvage (MS) and salvage index (SI) calculated from AAR and FIS.
RESULTS The STR in PPCI+SBP group was significantly higher than that in PPCI group (73.04±24.25 vs. 63.30±22.14, P=0.036), the peak value of hsTNT was lower than that of the PPCI group (5.54±3.23 vs. 7.09±4.51, P=0.048), and FIS was smaller than that of the PPCI group [6.50 (3.00–15.00) vs. 15.00 (4.00–29.50), P=0.047]. MS [2.50 (0.125–9.750) vs. 1.00 (-1.30 to 3.50), P=0.023] and SI [27.95 (0.00–45.00) vs. 4.30 (-18.35 to 20.55), P=0.006] were larger than those in the PPCI group. The LVEF was higher than that of the PPCI group (56.53±7.42% vs. 53.27±9.09%, P=0.049) and the NT-proBNP level was lower than that of the PPCI group (567.02±988.82 vs. 1014.86±1259.38, P=0.048).
CONCLUSIONS SBP can alleviate MI/RI (MS and SI), decrease in myocardial infarction area (peak value of hsTNT and FIS), and improve myocardial reperfusion (MBG and STR) and cardiac function (LVEF and NT-proBNP), so it protects myocardium in patients with STEMI.
GW31-e0643
Liwei Liu1,2, Jiyan Chen1,2
1The Second School of Clinical Medicine, Southern Medical University, Guangzhou, 510515 Guangdong, China
2Department of Cardiology, Guangdong Provincial Key Laboratory of Coronary Heart Disease Prevention, Guangdong Cardiovascular Institute, Guangdong
OBJECTIVES The emergence of AHF after percutaneous coronary intervention (PCI) would lead to a seriously worse prognosis. However, there is few models of predicting post-PCI AHF for patients undergoing PCI. Therefore, we aim to establish a simple prediction model of postoperative AHF among patients following PCI.
METHODS Among 2091 patients undergoing PCI in a prospective observational cohort (PRECOMIN, ClinicalTrials.gov NCT01400295) were enrolled from January 2010 and October 2012. The primary endpoint was post-PCI AHF, which was defined according to a detailed history of symptoms, previous cardiovascular events, the evaluation of signs/symptoms of congestion and/or low perfusion by the physical test with further confirmation by specific investigations such as ECG, chest X-ray, laboratory test (with cardiac biomarkers), and echocardiography. Based on the univariate logistic regression and stepwise logistic regression, we developed a clinical prediction model for post-PCI AHF by successively removing nonsignificant covariates. The discrimination of the nomogram was assessed by the area under the receiver operating characteristic (AUROC) curve and calibration was assessed using the Hosmer-Lemeshow statistic. We internally validated the model by using 1000 bootstrap samples to assess the stability of the model and translated this model into a nomogram. To evaluate the clinical implication of nomogram, we also presented by the classification of the model predictions into low-risk, middle-risk, and high-risk categories.
RESULTS Overall, the prevalence of post-PCI AHF was 1.91% (n=40). 402 (19.2%) of all enrolled patients are females and median age is 63 years (IQR 55–72) years. The final prediction model included heart rate (HR), acute myocardial infarction (AMI), age and left ventricular ejection fraction (LVEF). This model had a high discriminatory ability for post-PCI AHF (AUC=0.845), with calibration (Hosmer-Lemeshow statistic 6.063 P=0.640). For the internal validation using 1000 bootstrap samples, the bootstrap-corrected C-statistic based on data from the development cohort was 0.831. And the incidence of post-PCI AHF in the cohort based on the nomogram showed the good consistency in all three risk classification groups.
CONCLUSIONS The simple model of post-PCI AHF performed stable performance and improved clinical implication in risk stratification for high-risk individuals. Using a nomogram, the risk of each patient developing post-PCI AHF can be estimated, which is beneficial for clinicians in making early clinical decisions and prompt intervention.
GW31-e0645
Liwei Liu1,2, Jiyan Chen1,2
1The Second School of Clinical Medicine, Southern Medical University, Guangzhou 510515 Guangdong, China
2Guangdong Provincial People’s Hospital affiliated with South China University of Technology, Guangzhou 510000, Guangdong, China
OBJECTIVES Contrast-induced acute kidney injury (CI-AKI) was the major complication in patients with chronic kidney disease (CKD) undergoing angiography and usually led to poorer prognosis. Previous studies have shown that different definitions of CI-AKI have different effects on the long-term prognosis of patients. It is unknown which definition of CI-AKI accounts for most long-term mortality among patients with CKD. The population-attributable risk (PAR) represents the proportion of cases in a population that would not have occurred in the absence of a risk factor. To the best of our knowledge, no studies have quantified the contributions of different definitions of CIAKI to long-term mortality in patients with CKD. Therefore, we aimed to evaluate this association and compared the PARs of two major CI-AKI definitions.
METHODS This is a single-center, prospective observational study (PRECOMIN, NCT01400295) in Guangdong Provincial People’s Hospital, China. From January 2010 to December 2013, we enrolled patients aged ≥18 years who continued to be hospitalized for 2–3 days after angiography. And 610 consecutive coronary artery disease (CAD) patients with CKD undergoing angiography were included in the final analysis. The endpoint was all-cause mortality. CI-AKI was evaluated according to two major definitions: (1) CI-AKIA, with a serum creatinine elevation ≥50% or ≥0.3 mg/dL from baseline in the 72 h after the contrast procedure; (2) CI-AKIB ≥0.5 mg/dL or ≥25% in 72 h after the contrast procedure. Kaplan-Meier analysis was used to count the cumulative mortality, and the log-rank test was used to assess the differences between curves. Multivariable Cox analysis was conducted to evaluate the association between CI-AKI and long-term mortality and expressed as the hazard ratio (HR). The adjusted risk factors were selected through univariable Cox regression or based on previous studies and clinical importance. Population attributable risks (PAR) was used to evaluate the impact of two different definitions of CI-AKI and was calculated using the equation PAR=P (HR-1)/[1+P (HR-1)], where P is the prevalence of CI-AKI under different definitions in our database.
RESULTS Among 610 CAD patients with CKD, 113 patients developed CI-AKIA, and 95 patients developed CI-AKIB after contrast exposure. About 27% (165/610) of patients were female, the mean age was 70.01±9.55 years, and the mean eGFR was 45.66±11.43 mL/min/1.73 mm2. During the median follow-up period of 6.3 (5.8; 7.6) years, the mortality rate of patients was 25.57% (156/610). Kaplan-Meier curves revealed that patients with CIAKI demonstrated poorer long-term prognosis than those without CI-AKI (log-rank test, P<0.001). After adjusting for heart rate, eGFR, acute myocardial infarction, age, left ventricular ejection fraction <40%, blood urea nitrogen, and the use of intra-aortic balloon pump, both two definitions of CI-AKI was independently associated with the poorer outcome. Among the two definitions of CI-AKI, the adjusted HR of CI-AKIB was higher than CI-AKIA (1.862 vs. 1.817). And the prevalence of CI-AKIA was higher (18.5 vs. 15.6%), with the higher PAR (13.1%, 95% CI: 3.7–24.2).
CONCLUSIONS Our results suggested that CI-AKI is associated with long-term mortality in CAD patients with CKD irrespective of its definitions. Cardiologists and studies regarding long-term prognosis should pay more attention to the presence of CI-AKI, especially CI-AKIA with the higher PAR.
GW31-e0646
Liwei Liu1,2, Jiyan Chen1,2
1The Second School of Clinical Medicine, Southern Medical University, Guangzhou 510515 Guangdong, China
2Guangdong Provincial People’s Hospital affiliated with South China University of Technology, Guangzhou 510000, Guangdong, China
OBJECTIVES Contrast-induced acute kidney injury (CI-AKI) is a major adverse effect caused by intravascular administration of contrast medium. Current studies showed that hypoalbuminemia might be a novel risk factor for predicting CI-AKI. This study performed a systematic review and meta-analysis to investigate whether hypoalbuminemia is an independent risk factor for CI-AKI.
METHODS Relevant studies were searched in Ovid Medline, Embase, Cochrane Library until December 31, 2019. The inclusion and exclusion criteria were clearly addressed. Two authors independently screened studies for inclusion, consulting with a third author where necessary to resolve discrepancies. The pooled odds ratio was calculated to assess the association between hypoalbuminemia and risk of CI-AKI using a random-effects model or fix-effects model. Publication bias was tested using funnel plots and the Egger test.
RESULTS Eight relevant studies involving a total of 18,687 patients met our inclusion criteria. The presence of hypoalbuminemia was associated with an increased risk of CI-AKI development (pooled OR: 2.59, 95% CI: 1.80–3.73). It suggests that hypoalbuminemia should be evaluated to reduce the incidence of CI-AKI in patients undergoing contrast exposure.
CONCLUSIONS Hypoalbuminemia is independently associated with the occurrence of CI-AKI and may be a potentially modifiable risk factor for clinical intervention. However, more studies are needed to validate these findings.
GW31-e0652
Liwei Liu1,2, Jingjing Liang1,2,3, Jiyan Chen1,2
1The Second School of Clinical Medicine, Southern Medical University, Guangzhou 510515 Guangdong, China
2Department of Cardiology, Guangdong Provincial Key Laboratory of Coronary Heart Disease Prevention, Guangdong Cardiovascular Institute, Guangdong
3Department of Cardiology, Shunde Hospital of Southern Medical University, Shunde, China
OBJECTIVES Contrast-associated acute kidney injury (CA-AKI) is one of the major complications that occurs after contrast exposure and patients complicated with hypoalbuminemia are at high risk of CA-AKI. Patients with hypoalbuminemia are prone to CA-AKI and do not have their own risk stratification tool. Therefore, we developed and validated a nomogram for predicting CA-AKI in patients with hypoalbuminemia undergoing CAG/PCI.
METHODS A total of 1272 consecutive patients with hypoalbuminemia undergoing CAG/PCI were enrolled and randomly assigned (2:1 ratio) to a development cohort (n=848) and a validation cohort (n=424). CA-AKI was defined as a serum creatinine (SCr) increase of ≥0.3 mg/dL or 50% from baseline within the first 48–72 hours following CAG/PCI. A nomogram was established with independent predictors according to multivariate logistic regression and a stepwise approach. The discrimination of the nomogram was assessed by the area under the receiver operating characteristic (ROC) curve and was compared to the classic Mehran CA-AKI score. Calibration was assessed using the Hosmer–Lemeshow test.
RESULTS Overall, 8.4% (71/848) of patients in the development cohort and 11.2% (48/424) of patients in the validation cohort experienced CA-AKI. The simple nomogram included estimated glomerular filtration rate (eGFR), serum albumin (ALB), age and the use of intra-aortic balloon pump (IABP); showed better predictive ability than the Mehran score (C-index 75.6 vs. 69.3%, P=0.02); and had good calibration (Hosmer–Lemeshow test P=0.887). According to the log-rank analysis, patients with CA-AKI presented a worse long-term outcome.
CONCLUSIONS Our data suggested that the simple nomogram might be a good tool for predicting CA-AKI in high-risk patients with hypoalbuminemia undergoing CAG/PCI, but our findings require further external validation.
GW31-e0658
Mingkang Li1, Chengchun Tang2
1School of Medicine, Southeast University
2Department of Cardiology, Zhongda Hospital, Southeast University
OBJECTIVES Previous studies showed that fibrinogen-to-albumin ratio (FAR) regarding as a novel inflammatory and thrombotic biomarker was the risk factor for coronary artery disease (CAD). In this study, we sought to evaluate the relationship between FAR and severity of CAD, long-term prognosis in non-ST elevation acute coronary syndrome (NSTE-ACS) patients firstly implanted with drug-eluting stent (DES).
METHODS A total of 1138 consecutive NSTE-ACS patients firstly implanted with DES from January 2017 to December 2018 were recruited in this study. Patients were divided into tertiles according to FAR levels (Group 1: ≤8.715%; Group 2: 8.715–10.481%; Group 3: >10.481%). The severity of CAD was evaluated using the Gensini Score (GS). The endpoints were major adverse cardiovascular events (MACE), including all-cause mortality, myocardial reinfarction and target vessel revascularization (TVR).
RESULTS Positive correlation was detected by Spearman’s rank correlation coefficient analysis between FAR and GS (r=0.17, P<0.001). On multivariate logistic analysis, FAR was an independent predictor of severe CAD (OR=1.060, 95% CI 1.005~1.118, P<0.05). Multivariate Cox regression analysis indicated that FAR was an independent prognostic factor for MACE at 30 days, 6 months, and 1 year after DES implantation (HR 1.095, 95% CI 1.011~1.186, P=0.025; HR 1.076, 95% CI 1.009~1.147, P=0.026; HR 1.080, 95% CI 1.022~1.141, P=0.006, respectively). Furthermore, adding FAR to the model of established risk factors, the C-statistic increased from 0.706 to 0.720, 0.650 to 0.668, 0.611 to 0.632, respectively. And the models had incremental prognostic value for MACE, especially for 1-year MACE (NRI: 13.6% improvement, P=0.044; IDI: 0.6% improvement, P=0.042).
CONCLUSIONS In conclusion, FAR was associated independently with the severity of CAD and prognosis, helping to improve risk stratification in NSTE-ACS patients firstly implanted with DES.
GW31-e0667
Ming Ying1, Liwei Liu1,3, Jin Liu1, Yong Liu1,2,3, Haozhang Huang1,3, Zhujun Chen1
1Department of Cardiology, Provincial Key Laboratory of Coronary Heart Disease, Guangdong Cardiovascular Institute, Guangdong Provincial People’s Hospital, Affiliated Guangdong Provincial People’s Hospital of South China University of Technology, Guangzhou 510100, China
2Guangdong Provincial People’s Hospital, School of Medicine, South China University of Technology, Guangzhou 510000, China
3The Second School of Clinical Medicine, Southern Medical University, Guangzhou 510515, China
OBJECTIVES Contrast-induced acute kidney injury (CI-AKI) is one of the major complications after coronary angiography (CAG) or percutaneous coronary intervention (PCI). Nowadays, most of the risk factors predicting CI-AKI are irreversible and complicated, such as diabetes, congress heart failure and chronic kidney disease. Albumin is the most abundant circulating protein that plays an important role in anti-inflammatory and antioxidant activities. Serum albumin levels are strongly associated with cardiovascular morbidity and mortality. Serum albumin levels are strongly associated with cardiovascular morbidity and mortality. We investigated the relationship between Serum albumin and accidence of CI-AKI in patients after coronary angiography.
METHODS Two hundred eighty-nine consecutive patients aged ≥18 years who agreed to stay in the Guangdong Province’s Peoples Hospital 2–3 days after coronary angiography were consecutively enrolled from January 2010 till October 2012. According to whether patients developed contrast induced acute kidney injury or not, they were assigned to either a contrast-induced acute kidney injury group (80 cases, 2.69%) or a non-contrast-induced acute kidney injury group (2899 cases; control).
RESULTS On multivariate logistic regression analysis, adjusted with Chronic heart failure (OR 1.947, 1.103–3.407), Age (OR 1.268, 1.009–1.104) and IABP (OR 4.238, 2.007–8.750), Serum album (OR 0.860, 0.806–0.917) were independent predictors for CI-AKI in patients after coronary angiography. Levels of serum albumin ≤32.15 pg/mL (0.772, 0.72–0.83) were the optimal cutoff value. Serum Album is more effective in predicting CI-AKI in CKD patients (OR 0.809, 0.752–0.870) than in Non-CKD patients (OR 0.848, 0.786–0.914).
CONCLUSIONS Serum albumin was an independent risk factor of contrast-induced acute kidney injury among patients that received percutaneous coronary intervention. Album is more effective in predicting CI-AKI in CKD patients than in Non-CKD patients.
GW31-e0685
Haoyu Wang, Kefei Dou, Bo Xu, Yuejin Yang
Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College
OBJECTIVES The relation between complex percutaneous coronary intervention (PCI), high bleeding risk (HBR), and adverse events after coronary artery implantation of drug-eluting stents has been incompletely characterized. This study sought to investigate the ischemic and bleeding events after complex PCI including stratification according to HBR estimated by PARIS bleeding risk score.
METHODS Between January 2013 and December 2013, 10,167 consecutive patients undergoing PCI were prospectively enrolled in Fuwai PCI Registry. Complex PCI was defined when having at least one of the following characteristics: 3 vessels treated, ≥3 stents implanted, ≥3 lesions treated, bifurcation with 2 stents implanted, total stent length >60 mm, treatment of chronic total occlusion, unprotected left main PCI, in-stent restenosis target lesion, and severely calcified lesion. The primary ischemic endpoint was major adverse cardiovascular events (MACE) (composite of cardiac death, myocardial infarction, definite/probable stent thrombosis, and target lesion revascularization), and primary bleeding endpoint was Bleeding Academic Research Consortium (BARC) type 2, 3, or 5 bleeding.
RESULTS The median duration of follow-up was 29 months. In adjusted Cox regression analysis, patients having complex PCI procedures experienced higher risks of MACE (hazard ratio (HR): 1.63, 95% confidence interval (CI): 1.38–1.92; P<0.001), compared with noncomplex PCI. In contrast, the risk of clinically relevant bleeding was statistically similar between the 2 groups (HR: 0.86 [0.66–1.11]; P=0.238). There was no statistical interaction between HBR (PARIS bleeding score ≥8 or <8) and complex PCI in regard to MACE (adjusted P interaction=0.388) and clinically relevant bleeding (adjusted P interaction=0.279).
CONCLUSIONS Patients who had undergone complex PCI resulted in substantially more ischemic events, without an increase in clinically relevant bleeding risk, and these associations did not seem to be modified by HBR status. More intensified antiplatelet therapy may be beneficial for patients with complex percutaneous coronary revascularization procedures.
GW31-e0686
Haoyu Wang, Kefei Dou, Bo Xu, Runlin Gao
Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College
OBJECTIVES Whether the underlying risk of high bleeding risk (HBR) influences the relationship of high thrombotic risk (HTR) features with adverse events after drug-eluting stent implantation remains unclear. The purpose of this study was to evaluate (1) the prognostic effect of ESC guideline-endorsed HTR features on long-term clinical outcomes and (2) whether the outcomes of HTR versus non-HTR features vary by HBR status.
METHODS Of 10,167 consecutive patients who underwent percutaneous coronary intervention between January 2013 and December 2013 were prospectively enrolled in Fuwai PCI Registry. Patients who are at HTR were defined as: diffuse multivessel disease in diabetic patients, chronic kidney disease, at least three stents implanted, at least three stents lesions treated, bifurcation with two stents implanted, total stent length >60 mm, or treatment of chronic total occlusion. The definition of HBR was based on the Academic Research Consortium for HBR criteria. The primary ischemic outcome was major adverse cardiac event (MACE), a composite of cardiac death, myocardial infarction, target vessel revascularization and stent thrombosis. The primary bleeding outcome was clinically relevant bleeding, defined according to Bleeding Academic Research Consortium (BARC) type 2, 3 or 5 bleeding.
RESULTS With a 2.4-year median follow-up, 4430 patients (43.6%) having HTR experienced a significantly higher risk of MACE (hazard ratio [HR]adjust: 1.56, 95% confidence interval [CI]: 1.34–1.82; P<0.001) and device-oriented composite endpoint (composite of cardiac death, target-vessel MI, and target lesion revascularization) (HRadjust: 1.52 [1.27–1.83]; P<0.001), compared to those having non-HTR. The risk of clinically relevant bleeding did not differ between groups (HRadjust: 0.85 [0.66–1.08]; P=0.174). Associations between HTR and adverse events were similar in HBR and non-HBR groups, without evidence of interaction (all Pinteraction>0.05); however, adverse event rates were highest among subjects with both HTR and HBR.
CONCLUSIONS ESC guideline-endorsed HTR was associated with significantly increased risk of MACE without any significant differences in clinically relevant bleeding. The presence of HBR does not emerge as a modifier of cardiovascular risk for patients at HTR, suggesting more potent and longer antiplatelet therapy may be beneficial for this patient population.
GW31-e0687
Haoyu Wang, Bo Xu, Kefei Dou, Runlin Gao
Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College
OBJECTIVES The benefits and harms of dual antiplatelet therapy (DAPT) continuation with aspirin and clopidogrel beyond 1 year after percutaneous coronary intervention (PCI) with drug-eluting stent (DES) implantation for high ischemic or bleeding risk patients remain unclear.
METHODS All consecutive patients undergoing PCI were prospectively included in the Fuwai PCI Registry from January 2013 to December 2013. We evaluated 7521 patients who were at high risk for thrombotic or hemorrhagic complications and were events free at 1 year after the index procedure. “TWILIGHT-like” patients with high risk of bleeding or ischemic events were defined by clinical and angiographic criteria. The primary ischemic outcome was major adverse cardiac and cerebrovascular events [MACCE] (a composite of all-cause death, myocardial infarction, or stroke). Median follow-up duration was 2.4 years.
RESULTS The risk of MACCE was significantly lower in DAPT>1-year group (n=5252) than DAPT=1-year group (n=2269) (1.5 vs. 3.8%; hazard ratio [HR]: 0.37; 95% confidence interval [CI]: 0.27–0.50; P<0.001). This difference was largely driven by a lower risk of all-cause death. In contrast, the risk of Bleeding Academic Research Consortium (BARC) type 2, 3 or 5 bleeding was statistically similar between the 2 groups (1.0 vs. 1.1%; HR: 0.80; 95% CI: 0.50–1.28; P=0.346). Results were consistent after multivariable regression and propensity-score matching.
CONCLUSIONS Prolonged DAPT beyond 1 year after DES implantation resulted in a significantly lower rate of atherothrombotic events, including a mortality benefit, with no higher risk of clinically relevant bleeding in “TWILIGHT-like” patients who were at high-risk for ischemic or bleeding events.
GW31-e0688
Haoyu Wang, Kefei Dou, Bo Xu, Runlin Gao
Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College
OBJECTIVES The ischemic/bleeding trade-off of continuing dual antiplatelet therapy (DAPT) beyond 1 year after PCI for patients with high thrombotic risk (HTR) as endorsed by 2018 ESC/EACTS myocardial revascularization guidelines remain unknown.
METHODS Patients undergoing coronary stenting between January 2013 and December 2013 from the prospective Fuwai registry were defined as HTR if they met at least 1 ESC/EACTS guideline-endorsed HTR criteria. 4578 patients who were at HTR and were events free at 1 year after the index procedure were evaluated. The primary efficacy outcome was major adverse cardiac and cerebrovascular events [MACCE] (composite of all-cause death, myocardial infarction, or stroke).
RESULTS Median follow-up period was 2.4 years. >1-year DAPT with clopidogrel and aspirin significantly reduced the risk of MACCE compared with ≤1-year DAPT (1.9 vs. 4.6%; hazard ratio [HR]: 0.38; 95% confidence interval [CI]: 0.27–0.54; P<0.001), driven by a reduction in all-cause death (0.2 vs. 3.0%; HR: 0.07; 95% CI: 0.03–0.15). Cardiac death and definite/probable stent thrombosis also occurred less frequently in prolonged DAPT group. Bleeding Academic Research Consortium (BARC) type 2, 3 or 5 bleeding occurred similarly between both groups (1.1 vs. 0.9%; HR: 1.11; 95% CI: 0.58–2.13; P=0.763). Similar results were found using multivariable Cox model, propensity score-matched, and inverse probability of treatment weighting analysis.
CONCLUSIONS Among patients with ESC-endorsed HTR who were free from major ischemic or bleeding events 1 year after coronary stenting, continued DAPT beyond 1 year might offer better effectiveness in terms of atherothrombotic events and comparable safety in terms of clinically relevant bleeding compared with ≤1-year DAPT. ESC-HTR criteria is an important parameter to take into account in tailoring DAPT prolongation.
GW31-e0690
Chuyi Han1,2, Hongliang Cong1
1Tianjin Chest Hospital
2Tianjin Medical University
OBJECTIVES To evaluate the value of three systems, namely CRUSADE, PARIS, and PRECISE-DAPT, in predicting the risk of in-hospital hemorrhage in ACS patients receiving PCI.
METHODS We retrospectively analyzed 1384 consecutive patients with ACS received PCI attending. In this study, End point of bleeding event was defined as BARC non-CABG-related bleeding ≥type 2 (except type 4). The predictive value of three scoring systems for nosocomial bleeding events after intervention was evaluated by drawing a ROC curves.
RESULTS (1) In the selected 2 278 patients diagnosed with ACS, 1384 were treated with PCI and finally enrolled after applying the exclusions. 252 patients of the 1384 patients had non-CABG-related bleeding events of type BARC ≥2 (except type 4). There were 128 cases of type 2, 114 cases of type 3 and 10 cases of type 5. (2) Comparison of bleeding scores among three series in bleeding group When BARC ≥type 2 (except type 4) was used as the end point of hemorrhage, CRUSADE score (32.78±13.89), PARIS score (5.02±2.26), and PRECISE-DAPT score (18.88±10.86) of patients with hemorrhage were significantly higher than those without hemorrhage (22.36±12.45), (3.82±1.72), and (12.69±7.51) (all P<0.001), respectively. (3) Logistic regression analysis of the bleeding event as the dependent variable. According to the CRUSADE score, the patients were divided into five groups. Logistic regression analysis showed that the risk of bleeding increased along with the increase of the CRUSADE score. The risk of bleeding in group B was 2.031 times higher than in group A. Group C was 2.047 times higher than in group A. Group D was 2.441 times higher than in group A, and group E was 4.885 times higher than in group A. According to the PARIS score, the patients were divided into three groups. Logistic regression analysis showed that the risk of bleeding increased with the increase of PRECISE-DAPT score. The risk of bleeding in group B was 1.801 times higher than in group A, and the risk of bleeding in group C was 5.053 times higher than in group A. According to the PRECISE-DAPT score, the enrolled patients were divided into four groups. Logistic regression analysis showed that the risk of bleeding increased with the increase of PRECISE-DAPT score. The risk of bleeding in group B was 1.434 times higher than in group A. The risk of bleeding in group C was 2.452 times higher than in group A, and the risk of bleeding in group D was 4.267 times higher than in group A. (4) ROC curve analysis on predictive ability for bleeding events. Using BARC ≥type 2 (except type 4) as the bleeding criteria, the area under the curve of the CRUSADE, PARIS and PRECISE DAPT score was 0.694 (95% CI: 0.669–0.718), 0.660 (95% CI: 0.634–0.685), 0.679 (95% CI: 0.654–0.704). The results showed that all three scores are valuable on the assessment of in-hospital hemorrhage in patients with ACS after PCI. No differences were observed in the ability of the three scores to predict nosocomial bleeding events in patients with ACS.
CONCLUSIONS All three scores have predictive value for nosocomial bleeding events in the patients of our study. They can be used for hazard layering. No differences were observed in the ability of the three scores to predict nosocomial bleeding events in patients with ACS.
GW31-e0697
Donglei Luo, Jingtao Guo
Chengde Central Hospital/Second Clinical College of Chengde Medical University
OBJECTIVES To explore the clinical significance of Hoffmayer ECG integral method in differential idiopathic right ventricular ventricular premature beat (PVC) or ventricular tachycardia (VT) with arrhythmogenic right ventricular cardiomyopathy (ARVC) PVC or VT in early stage
METHODS Forty cases with heart disease patients were collected, who were from the Chengde Central Hospital from Sep. 2016 to Sep. 2019. In all of them, 30 cases with right ventricular outflow tract PVC or VT, 10 cases with ARVC sinus rhythm concomitant PVC or VT. And then the ECG integrals of 40 patients were analysed through the calculating of the total and single integral of the patients with ARVC concomitant PVC or VT, in the meanwhile, the clinical diagnosis was compared. The susceptibility, specificity, positive and negative predictive value and diagnosis coincidence rate of the patients were observed.
RESULTS The susceptibility, specificity, positive and negative predictive value and diagnosis coincidence rate of the patients was 80, 90, 80, 81.3 and 90% respectively through the Hoffmayer ECG integral for the patients with ARVC concomitant PVC or VT was more than 5 points or less than 5 points.
CONCLUSIONS It’s effective to differential diagnose idiopathic right ventricular ventricular premature beat or ventricular–tachycardia with arrhythmogenic right ventricular cardiomyopathy concomitant PVC or VT. It’s worth applying in clinical for the higher sentivity and speciality, fastly and simply.
GW31-e0727
Jingjing Xu, Jinqing Yuan
Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Science and Peking Union Medical College
OBJECTIVES The definite effect of metabolic syndrome on the onset age and long-term outcome in acute coronary syndrome (ACS) patients remains unclear. The aim of this study is to investigate the effect of metabolic syndrome on onset age and long-term outcome of ACS patients.
METHODS Of 6431 ACS patients undergoing PCI from January to December 2013 were enrolled. After the exclusion of previous coronary artery disease history, 1558 patients accorded with early-onset ACS (man ≤50 and woman ≤60 years old), and 3044 patients with late-onset ACS. The baseline characteristics and 5 years clinical outcomes were collected.
RESULTS Compared with late-onset ACS group, BMI, levels of TG, LDL-C, and uric acid were significantly higher, while HDL-C was lower in early-onset ACS group (all P<0.001). Multivariable logistic analysis showed metabolic syndrome components including obesity (odd ratio: 1.590, P<0.001), hypertriglyceridemia (odd ratio: 1.403, P<0.001) and low HDL-C (odd ratio: 1.464, P<0.001) were independent risk factors of early-onset ACS. Five years follow-up showed that, the incidence of all cause death (1.5 vs. 3.8%, P<0.001), cardiac death (1.1 vs. 2.0%, P=0.023) and recurrent stroke (2.2 vs. 4.2%, P<0.001) were lower, while the bleeding events were higher (16.4 vs. 12.4%, P<0.001) in early-onset ACS group. However, subgroup analysis showed a higher incidence of recurrent MI and revascularization in early-onset ACS patients combined with metabolic syndrome.
CONCLUSIONS Metabolic syndrome components including obesity, hypertriglyceridemia and low HDL-C are independent risk factors of early-onset ACS and relate to the increase of recurrent MI and revascularization. Effective control of metabolic syndrome may reduce the incidence of early-onset ACS and improve long-term prognosis.
GW31-e0740
Sida Jia, Jianxin Li, Ce Zhang, Yue Liu, Deshan Yuan, Na Xu, Xueyan Zhao, Runlin Gao, Yuejin Yang, Bo Xu, Zhan Gao, Jinqing Yuan, Yin Zhang
Fuwai Hospital, Chinese Academy of Medical Science
OBJECTIVES Moderate/severe coronary calcification predicts worse clinical outcomes in patients undergoing percutaneous coronary intervention (PCI). However, to date most studies were modest in size and with limited follow-up. We aim to assess the association between calcification severity and long-term clinical outcomes in a large patient cohort undergoing PCI.
METHODS Totally 10,068 consecutive patients who underwent PCI at Fuwai Hospital were enrolled in this prospective observational study. Patients were categorized as none/mild calcification or moderate/severe calcification according to the severity of target lesion by visual assessment of coronary angiography. Major adverse cardiovascular events (MACE), a composite event of death, myocardial infarction and revascularization, at 5 years were assessed.
RESULTS None/mild calcification was observed in 8229 (81.7%) patients, while moderate/severe calcification was observed in 1839 (18.3%) patients. Patients with moderate/severe calcification had significantly higher rate of 5-year unplanned revascularization (15.2 vs. 13.2%, P=0.022) and MACE (20.7 vs. 17.9%, P=0.005). After propensity score match, moderate/severe CAC group still had higher rate of 5-year unplanned revascularization (15.2 vs. 12.6%, P=0.019). Multivariable Cox regression analysis found that moderate/severe calcification was independently associated with higher risk of 2-year unplanned TVR (HR=1.287, 95% CI: 1.036–1.600, P=0.023) and MACE (HR=1.242, 95% CI: 1.039–1.484, P=0.017), but not 5-year unplanned revascularization and MACE.
CONCLUSIONS In patients undergoing PCI, moderate/severe coronary calcifications increases the risk of long-term MACE.
GW31-e0770
Yuting Liu, Qingshan Geng
Guangdong Cardiovascular Institute, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong, China
OBJECTIVES The aim of this study was to explore the changes of heart rate variability, salivary cortisol and salivary α-amylase in CDA disease group during mental stress comparing with healthy control group and CHD group.
METHODS The Patient Health Questionnaire-9 (PHQ-9) and Generalized Anxiety Disorder-7 (GAD-7) were used to evaluate anxiety and depression disorders in patients with CHD (N=81) and healthy individuals (N=10). According to the scale score, the subjects were divided into 3 groups: the healthy control group, the CHD group and the CDA group (CHD comorbid mild and above anxiety and/or depression), and all subjects were evaluated for changes in hemodynamics, heart rate variability, salivary cortisol and salivary α-amylase during the mental stress test, followed by subjective psychological evaluation. Statistical comparison analysis was performed between different time points and between groups. correlation factor analysis was performed. Correlation analysis was carried out on the factors that may affect the mental stress responses.
RESULTS The autonomic nerve response to mental stress in CDA patients was characterized by low reactivity and slow recovery, with no significant changes in rMSSD and HF during mental stress, less increase in LFnu than that of the healthy and CHD groups (31.5 vs. 68.5 vs. 33.9%) and with delayed recovery to baseline. The responses of hypothalamic-pituitary-adrenal axis and the sympathetic-adrenal medulla system under mental stress have similar trends to those in healthy individuals and CHD groups, showing a peak at 10 minutes after mental stress and a return to baseline 30 minutes after. After adjusting for other factors, the correlation between high levels of GAD-7 in CDA patients and low response in sympathetic nerve activity was still significant (F=6.528, P=0.014), showing a progressive low response to stressful events. Besides, the subjective stress score was found to be significantly positively correlated with changes of most indicators in the response to mental stress.
CONCLUSIONS The physiological flexibility of patients with CDA is diminished under mental stress, which is manifested by a significant reduction in the amplitude of autonomic nerve responses, which is also closely related to the degree of anxiety and subjective stress feeling.
GW31-e0775
Juan Zhou, Zuyi Yuan
Department of Cardiology, First Affiliated Hospital, Xi’an Jiaotong University
OBJECTIVES To describe long-term antithrombotic management patterns (AMPs) in medically managed Asian patients with non-ST-segment myocardial infarction (NSTEMI) or unstable angina (UA).
METHODS Data were analyzed from medically managed NSTEMI and UA patients included in the prospective, observational EPICOR Asia study (NCT01361386). Survivors to hospital discharge were enrolled (June 2011 to May 2012) from 8 countries/regions across Asia. Baseline characteristics and AMP use up to 2 years post-discharge were collected. Outcomes were major adverse cardiovascular events (MACE: myocardial infarction, ischemic stroke, and death) and bleeding.
RESULTS Among 2289 medically managed patients, dual antiplatelet therapy (DAPT) use at discharge was greater in NSTEMI than in UA patients (81.8 vs. 65.3%), and was significantly associated with male sex, positive cardiac markers, and prior cardiovascular medications (P<0.0001). By 2 years, 57.9 and 42.6% of NSTEMI and UA patients, respectively, were on DAPT. On multivariable Cox regression analysis, risk of MACE at 2 years was most significantly associated with older age (HR [95% CI] 1.85 [1.36, 2.50]), diagnosis of NSTEMI vs. UA (1.96 [1.47, 2.61]), and chronic renal failure (2.14 [1.34, 3.41]), all P=0.001. Risk of bleeding was most significantly associated with region (East Asia vs. Southeast/South Asia) and diabetes.
CONCLUSIONS Approximately half of all patients were on DAPT at 2 years. MACE were more frequent in NSTEMI patients during follow-up.
GW31-e0776
Xin Su1,2, Daoquan Peng2
1Xiamen Cardiovascular Hospital of Xiamen University
2Second Xiangya Hospital of Central South University
OBJECTIVES Guidelines recommended non-high density lipoprotein cholesterol (non-HDL-C) as a co-primary target, and set non-HDL-C goals as 30 mg/dL higher than low density lipoprotein cholesterol (LDL-C) goals. However, the value is largely uncertain in Chinese patients.
METHODS We assigned non-HDL-C values at the same percentiles correspondent to LDL-C goals for patients from the Improving Care for Cardiovascular Disease in China-Acute Coronary Syndrome (CCC-ACS) Project. We calculated the differences between non-HDL-C and LDL-C and proposed appropriate adding values according to LDL-C and TG levels.
RESULTS Among 73,495 patients, 17.7% used lipid-lowering agents before admission. Of these, 27.2% achieved LDL-C<70 mg/dL while 39.4% achieved non-HDL-C<100 mg/dL. The mean difference between non-HDL-C and LDL-C was 23.2 mg/dL, which could be affected by LDL-C and TG levels. Importantly, of patients with LDL-C levels ≤100 mg/dL, the mean differences were 19.1 mg/dL in patients with TG ≤150 mg/dL and 24.6 mg/dL in patients with TG >150 mg/dL.
CONCLUSIONS There are significant differences between LDL-C and non-HDL-C in Chinese ACS patients. For secondary prevention, on average, the adding values should be 20 mg/dL for patients with TG ≤150 mg/dL and 25 mg/dL for patients with TG >150 mg/dL when LDL-C goals of 70 mg/dL is achieved.
GW31-e0782
Dongxia Jin1,2, Hongliang Cong1,2, Tingting Li1,2, Ximing Li1,2
1Tianjin Chest Hospital
2Institute of Cardiovascular Disease of Tianjin in China
OBJECTIVES Studying the current states of contrast-induced encephalopathy (CIE) of atherosclerotic cardiovascular disease (ASCVD) in documents, including the clinical epidemiological characteristics, pathogenic mechanism and clinical symptoms, and analyzing the patients’ gender, age, preexisted hypertension, diabetes mellitus, old cerebral infarction (OCI) or transient ischemic attack (TIA), renal impairment or renal dysfunction, the category and doses of contrast, manifestation and prognosis, so as to promote the diagnosis and treatment of CIE of ASCVD.
METHODS Search the paper with Chinese and English in database. The retrieval words are “atherosclerotic cardiovascular disease, contrast-induced encephalopathy, neurotoxicity, encephalopathy, brain injury, cortical blindness, cerebral/carotid arteriography, cardiac catheterization, coronary angiography, percutaneous coronary intervention, peripheral arteriography”. Consult the documents and the references from the primary articles. And then bring in the cases of CIE of ASCVD, referred to the diagnostic criteria of NLA in 2014, and then collect the basic information and use the SPSS 20.0 and Excel statistic software to conduct the retrospective analysis.
RESULTS In the total 90 cases including 88 patients, male patient is 58, occupying 65.91%, while female is 30, accounting for 34.09%. It seems that there is a higher incidence of CIE in male patient. Age conforms to normal distribution, and the minimal age is 39, the maximal is 89, the average is (66.88±10.51) years old, which indicates that possibly the elder suffers more from CIE. And the number of hypertension is 63 (71.59%), and non-hypertension is 25 (28.41%). The mean and median volumes of iodinated contrast media administered were 228.41 and 190 mL, respectively (range: 25–1500 mL). High-, low-, iso-osmolar contrast media were used in 8.89% (8/90), 74/90 (82.22%) and 1/90 (1.11)% of patients, respectively. Symptoms of patients: cortical blindness, delirium, hemiplegia, seizure, ophthalmoplegia accounted for 25.56, 13.33, 10.00, 8.89 and 5.56% respectively. Symptoms typically appear within minutes to hours of contrast administration and resolve entirely within 24–72 hours. And most of the patients (86/90, about 95.56%) had completely recovered, and only 4 patients suffered irreversible damage: 1 suffered of right-hand muscle strength decline, 1 suffered of blepharoptosis, diplopia, and 1 with persistent forgetting and vision loss, unfortunately 1 patient died from CIE after 56-days treatment. Risk factors may include hypertension, renal impairment, transient ischemic attack, the administration of large volumes of iodinated contrast, percutaneous coronary intervention or selective angiography of internal mammary grafts.
CONCLUSIONS Though CIE is a very rare complication during the intervention procedure and the presentation of CIE is variable, ranging from cortical blindness to encephalopathy, seizures, and hemiparesis, which often pose a difficult differential diagnosis. In any acutely confused patient, it is important, however, to carry out the relevant investigations, as CIE is often a diagnosis of exclusion. Usually its prognosis is excellent, rarely can it bring some clinical troubles with the neurological sequelae, even to death. Physicians should be aware of it in time and take some measures to treat it.
GW31-e0817
Haoyu Wu
Department of Cardiology, Shaanxi Provincial People’s Hospital, Shaanxi, Xi’an, China
OBJECTIVES Although there have been many studies on circulating homocysteine and the incidence of coronary heart disease (CHD). However, the result on homocysteine (Hcy) in CHD has been conflicting as several studies have failed to demonstrate an association between Hcy and CHD, especially in Asian population. These studies include a small population. Therefore, this study aims to assess the relationship between Hcy level and CHD in the northern communities of China. We compared the adjusted conventional risk factors with Hcy in CHD patients to determine the predictive rate of CHD.
METHODS A total of 1987 patients in northern China who had undergone coronary angiographies were enrolled in the study, of which 672 with normal coronary arteries (control group) and 1315 proven CHD. Hcy levels were measured by enzyme-linked immunosorbent assay. Hyperhomocysteinemia (HHcy) was defined as Hcy ≥15 μmol/L.
RESULTS There were significant differences between the CHD and control groups with regard to male sex (P<0.05), diabetes (P<0.001), hypertension (P<0.001) and smokers (P<0.001), but no remarkable difference in family history of premature CHD (P>0.05). Hcy levels were significantly higher in the CHD group than those in the control group (P<0.001). Multivariate logistic regression showed that HHcy were both independently correlated with CHD in young patients (aged ≤55 years, OR, 3.03; 95% CI, 2.10–5.76) and old patients (aged >55 years, OR, 2.56; 95% CI, 1.21–4.25). Male, diabetes, hypertension, and smoking were also independently correlated with CHD in young and old patients. HHcy showed the highest sensitivity (93.05%), specificity (86.12%), positive prediction value (89.94%), negative prediction value (90.56%) and accuracy (90.01%) when compared to other risk factors (area under curve: 0.904, 95% CI: 0.865–0.924, P<0.001). Cut-off values of 15.13 μmol/L for Hcy (Youden’s index=0.815) gave the highest index.
CONCLUSIONS HHcy is an important independent risk factor for CHD in northern China after adjusting for other risk factors.
GW31-e0818
Haoyu Wu
Department of Cardiology, Shaanxi Provincial People’s Hospital, Xi’an 710068
OBJECTIVES To investigate the effect of Gelan Xinning soft capsule on the expression of insulin-like growth factor-1 (IGF-1) in patients with unstable angina pectoris.
METHODS Ninety-two patients with unstable angina pectoris were randomly divided into control group and observation group. The control group was given conventional drug treatment, and the observation group was given Gelan Xinning soft capsule based on conventional drug treatment, 2 capsules/time, 3 times/day. The course of treatment was 8 weeks. The clinical efficacy and serum IGF-1 level were observed.
RESULTS The frequency and duration of angina pectoris in the observation group and the control group after treatment were decreased compared with those before treatment (P<0.05). The frequency and duration of angina pectoris in the observation group were lower than those in the control group after treatment (P<0.05). The serum IGF-1 level in the observation group and the control group after treatment was higher than that before treatment (P<0.05). The serum IGF-1 level in the observation group was significantly higher than that in the control group after treatment (P<0.05).
CONCLUSIONS Gelan Xinning soft capsule has good therapeutic effect on unstable angina pectoris, which may be related to the increase of IGF-1 level.
GW31-e0819
Haoyu Wu
Shaanxi Provincial People’s Hospital, Xi’an 710068, Shaanxi, China
OBJECTIVES To investigate the relative factors of in-stent restenosis (ISR) in patients with mild to moderate chronic kidney disease after coronary drug-eluting stent implantation.
METHODS One hundred and eighty-nine cases of patients with mild to moderate chronic kidney disease were performed coronary drug-eluting stent from November 2015 to September 2018. According to the second angiography results performed 9~15 months after stent implantation, patients were divided into ISR group and non-ISR group. Clinical data and biochemical indicators were analyzed.
RESULTS The mean follow-up time was (12.6±2.1) months, and the incidence of ISR was 20.1%. Compared with the non-ISR group, active smoking, family history of myocardial infarction, diabetes mellitus, glycosylated hemoglobin, low density lipoprotein, eGFR and uric acid were statistically significant in ISR group (P<0.05). Multivariate logistic proportional hazards regression model showed that diabetes mellitus, glycosylated hemoglobin, uric acid and family history of myocardial infarction were independent risk factors for ISR.
CONCLUSIONS Diabetes mellitus, family history of myocardial infarction, glycosylated hemoglobin and uric acid are independent risk factors of ISR in patients with mild to moderate chronic kidney disease after coronary drug-eluting stent implantation.
GW31-e0850
Lixue Xu, Yi He
Capital Medical University Affiliated Beijing Friendship Hospital
OBJECTIVES To explore impact of patient related, vessel related, image quality related and cardiovascular risk factors on CCTA interpretability by using a 256-detector row CT.
METHODS One hundred and ten patients who underwent CCTA and invasive coronary angiography (ICA) were consecutively retrospectively enrolled from January, 2018 to October, 2018. Using ICA as the reference standard, ≥50% diameter stenosis was defined as the cutoff criterion to detect diagnostic capacity of CCTA. Diagnostic consistency was investigated by calculating interrater reproducibility of CCTA. Multiple logistic regression models were performed to evaluate impact of 14 objective factors.
RESULTS A total of 1019 segments were evaluated. Per-segment sensitivity, specificity, accuracy, positive predictive value, negative predictive value of CCTA was 76.8, 93.7, 91.2, 67.8 and 95.9%, respectively. Per-segment diagnostic consistency was 0.44 for CCTA. Discrimination and calibration of regression models were satisfied. For accuracy, negative association was found in stenosis severity, calcium load and hyperlipidemia. For sensitivity, calcium load and diabetes mellitus (DM) was positively correlated. For specificity, negative correlation was observed in stenosis severity and calcium load. For interrater reproducibility, stenosis severity and calcium load were negatively associated while male sex and signal to noise ratio (SNR) were positively related (all P<0.05).
CONCLUSIONS Per-segment 256-detector row CCTA capacity was best in no stenosis or occluded segments. Heavier calcium load was correlated to poorer CCTA interpretability. Our findings, on one hand, confirmed the rule-out value of CCTA, on the other hand, suggested improvements in calcium subtractions and deep learning-based tools to improve CCTA diagnostic interpretability.
GW31-e0851
Lixue Xu, Yi He
Capital Medical University Affiliated Beijing Friendship Hospital
OBJECTIVES Although 256-detector coronary CT angiography (CCTA) performed better in patients with higher heart rate, direct comparison of diagnostic performance between 64- and 256-detector row CCTA in patients with different pretest probability (PTP) of coronary artery disease (CAD) and in clinical important subgroups remained unknown.
METHODS One hundred and ten patients (76 males, 63.8 years±9.3 years) who underwent 256-detector row CCTA and 120 patients (68 males, 65.7±8.8 years) who underwent 64-detector row CCTA were retrospectively enrolled from January, 2017 to October, 2018. Multiple clinical information was recorded and applied to construct a risk model of CAD by conducting a multivariable regression model. The PTP of CAD was calculated for every patient, then used to stratify patients into PTP<15% and ≥15% risk subgroups. Using invasive angiography as reference standard and ≥50% stenosis as cutoff, group comparison of diagnostic performance was calculated between 64- and 256-detector row CCTA in risk-based, sex and age-based subgroups and in patients with and without chest pain.
RESULTS Sixty-four and 166 patients were with risk<15% and ≥15%, respectively. Group comparison showed that per-segment diagnostic performance of 256-detector row CCTA was significantly better than 64-detector row CCTA, especially in patients with risk of ≥15%, with age<65 years, with chest pain and in males.
CONCLUSIONS Our findings confirmed >64 detector row CCTA was more appropriate diagnostic test in chest pain center and for patients with PTP=15%.
GW31-e0855
Sulan Huang, Ning Guo
The First People’s Hospital of Changde
OBJECTIVES Some studies have showed that there was an independent association between increased red cell distribution width (RDW) and mortality after acute myocardial infarction (AMI). However, evidence regarding the predictive significance of repeated-measure of RDW in patients with AMI remains scarce. We investigated the association of dynamic profile of RDW and in-hospital mortality in a large population with AMI.
METHODS We extracted clinical data from the MIMIC-III V1.4 database. Then, we collected demographics, vital signs, laboratory tests, other clinical data and comorbidities from the database. The clinical endpoint was in-hospital mortality among critically ill patients with AMI. Cox proportional hazards models were used to evaluate the prognostic values of basic RDW, and the Kaplan-Meier method was used to plot survival curves. Subgroup analyses were performed to measure mortality across various subgroups. The repeated-measure data be compared by using generalized additive mixed model.
RESULTS A total of 3101 eligible patients were studied. In multivariate analysis, adjusted for age, ethnicity and gender, RDW were significant predictors of risk of in-hospital mortality. Furthermore, after adjusting for more confounding factors, RDW remained a significant predictor of in-hospital mortality (tertlile 3 versus tertile 1: adjusted HR, 95% CI: 2.36, 1.39–4.01; P for trend<0.05). A similar trend was observed in the RDW group division by quartiles. The Kaplan-Meier curve for tertile of RDW indicated that survival rates were the highest when RDW ≤13.2% and the lowest when RDW≥14.2% after adjustment for age, gender, and ethnicity. Subgroup analysis revealed no significant interactions in most strata. During staying in ICU, the RDW of most patients had raised up over time. The levels RDW of survivors changed smoothly and lower than those in non-survivors. Similarly, the levels RDW of non-survivors progressively increased after 3 days in ICU.
CONCLUSIONS Our findings showed that higher RDW was associated with risk of in-hospital mortality in critically ill patients with AMI.
GW31-e0915
Shuang Li1, Jingying Zhang1, Xiang Li2, Qiang Ming1, Xianlin Zhang3, Wei Chen1, Mengyun Zhu1, Yatyin Lam4, Yawei Xu1
1Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine
2Nursing, Shanghai Tenth People’s Hospital, Tongji University School of Medicine
3Cardiology, The First Affiliated Hospital of Bengbu Medical College
4Division of Cardiology Department of Medicine and Therapeutics, Clinical Sciences Building Prince of Wales Hospital, Hong Kong SAR
OBJECTIVES Percutaneous left atrial appendage (LAA) closure has been demonstrated to be an alternative to oral anticoagulation for the prevention of ischemic stroke in non-valvular atrial fibrillation (AF) patients using Watchman. However, few data are available for the novel LAmbre occluder, especially with long-term data. This study was to evaluate the feasibility of LAmbre LAA occluder followed for more than 5 years.
METHODS This study included consecutively 66 consecutive AF patients implanted with the LAmbre from April 2014 to October 2015 in our center. Patients received 3-month, double anti-platelet therapies and single alone thereafter. Patients were followed for up to July 7, 2020.
RESULTS The mean CHA2DS2-VASc and HAS-BLED score were 6.0±1.6 and 3.0±0.8, respectively. The implantation success rate was 98.5%. No death, stroke, major bleedings or device-related severe events were observed within 7 days after the procedure. Adequate LAA sealing was observed in 98.3 and 98.2% patients at respectively 3- and 12-month follow-up echocardiographic examinations. After a mean follow-up of 67±12.1 months, all-cause death was observed as 2.5 per 100 patient-years (n=9). Ischemic stroke and major bleeding were 1.9 per 100 patient-years (n=7) and 0.8 per 100 patient-years (n=3), respectively, both significantly lower than the cumulative expected rates according to risk factors.
CONCLUSIONS In patients with AF, the protocol of LAA closure with the LAmbre plus anti-platelet therapy was feasible and associated with low ischemic and bleeding events in the periprocedural period and in a long-term follow-ups of more than 5-years.
GW31-e0945
Jie Zhang, Likun Ma
Department of Cardiology, the First Affiliated Hospital of USTC, Division of Life Science and Medicine, University of Science and Technology of China, Hefei, Anhui, P.R. China
OBJECTIVES To evaluate the effect of NOAF after AMI on prognostic patients.
METHODS In this study, we evaluated 335 patients with AMI admitted to our department of cardiology with emergency coronary intervention during from January 2017 to December 2018. According to whether atrial fibrillation (AF) occurred during hospitalization, patients were classified as NOAF (n=32) group and no-NOAF (n=299) group. Baseline clinical characteristics (general clinical conditions, anamnesis, biochemical indexes, myocardial injure markers, NT-proBNP, cardiac color ultrasound, coronary angiography) of the two groups were counted and analyzed.
RESULTS We compared outcomes between these groups and adjusting for differences in baseline characteristics. After multivariate adjustment, NOAF was an independent predictor of death in-hospital (odds ratio (OR=4.259, P=0.024) rather than in 2-year (OR=3.061, P=0.124). In addition, NOAF was an independent predictor of adverse prognostic events like pump failure (OR=6.770, P=0.000), cardiogenic shock (OR=8.321, P=0.000), malignant arrhythmia (OR=2.877, P=0.027) in-hospital and heart failure (OR=9.892, P=0.004) in 2-year.
CONCLUSIONS These suggest that although NOAF was an independent predictor of all-cause death in-hospital but not in 2-year, it was associated with worse prognosis closely.
GW31-e0984
Jiaxin Cao, Yi He
Beijing Friendship Hospital, Capital Medical University
OBJECTIVES Image quality and diagnostic accuracy of coronary CT angiography (CCTA) could be impaired with inability of breath holding, high heart rate and heart rate variability. A novel dedicated cardiovascular CT system (CardioGraphe™) combining stereo dual source with 14 cm z-coverage and new iterative reconstruction algorithm (ASiR-CV) has come into clinical service recently. We aim to evaluate the feasibility of CCTA in patients with atrial fibrillation (AF) and free-breathing using the new dedicated cardiac CT scanner.
METHODS Thirty patients with AF (persistent AF without antiarrhythmic medication, 21 men and 9 women; mean age 70.3±9.4 years, ranged 40–82 years; mean BMI 26.8±3.4 kg/m2, ranged from 21.48 to 35.82 kg/m2) underwent clinically indicated CCTA on a 560-slice scanner (120 kV, 50-600 Smart mA, 0.24 s/rot, 560×0.25 mm collimation). Twelve patients were holding breath during acquisition time (group A) and in the remaining 18 with free-breathing (group B). A motion correction algorithm (SnapShot Freeze) was used to further minimize motion artifacts. A total of 316 in 538 coronary artery segments were rated as interpretable. Two experienced radiologists blinded to the electrocardiograph, independently graded the CT images in terms of visibility and artifacts with a 4-grade Likert rating scale. SNR and CNR were calculated by mean CT attenuation values and standard deviations within 3 regions of interest placed in the proximal left main (LM) and proximal right coronary artery (RCA). The CNR and SNR of two groups were compared using independent-sample T-test. The image quality of two groups were compared using Wilcoxon rank-sum test.
RESULTS Mean heart rate during scanning was 101.4±43.4 bpm in group A and 90.5±11.2 bpm in group B (P>0.05). The CNR and SNR of LM and RCA showed no significant difference between the two groups. There was no significant difference in image quality score between group A and group B on per-vessel level (3.7±0.4 vs. 3.6±0.5, P>0.05) and per-patient level (3.7±0.5 vs. 3.6±0.5, P>0.05).
CONCLUSIONS Free-breathing CCTA using the new dedicated cardiac CT scanner in patients with AF is equally effective with free-holding.
GW31-e1000
Yi Zhu, Naifeng Liu
Department of Cardiology, Zhongda Hospital, School of Medicine, Southeast University, Nanjing
OBJECTIVES Periostin (PN) is important for bone and tooth maintenance. CML (basis of advanced glycation end products) accelerates diabetic vascular calcification. This study aims to investigate the association between serum PN levels, CML levels and coronary artery Agatston score, and the effects of PN on vascular smooth muscle cell (VSMC) osteogenic differentiation in vitro.
METHODS We recruited 147 patients with suspected angina pectoris. Subjects were divided into 3 groups based on coronary Agatston score: score=0 (n=51), score=1–300 (n=39), score >300 (n=57). Serum PN and CML levels were measured using ELISA. VSMC calcification was induced with β-glycerophosphate (β-GP).
RESULTS Patients with coronary artery calcification (Agatston score >0) exhibited significantly higher serum PN and higher CML levels than those of control patients (Agatston score=0). Cardiovascular risk factors, including older age, male gender, lower creatinine clearance, diabetes mellitus and hypertension were more prevalent in patients with coronary calcification. High PN levels were associated with high CML levels. However, multivariate logistic analysis showed that high PN and CML levels could not predict the occurrence of coronary calcification. In vitro, PN elevated VSMC osteogenic differentiation marker (RUNX2 and BMP2) protein levels in a dose-dependent manner and that maximal stimulation was attained at 24 h.
CONCLUSIONS These findings suggest that diabetic vascular calcification is an important determinant of the induced circulating PN levels.
GW31-e1048
Xuelian Song, Yi Dang, Xiaoyong Qi
Hebei General Hospital
OBJECTIVES Acute myocardial infarction (AMI) is the most serious type of coronary heart disease that can be influenced by multiple environmental and inherited factors. It is well known that dyslipidemia is a definitely predisposing factor of acute myocardial infarction, further more, previous studies have found many single nucleotide polymorphisms (SNPs) of lipid metabolism associated with AMI occurrence. However, the results are inconsistent. So we designed this study to investigate four genes of lipid metabolism pathways by analysis of 4 SNPs that have not been reported the association with AMI in Han population of north China.
METHODS Three hundred and thirty-six patients with AMI confirmed by coronary angiography and 270 normal healthy controls are included. Four single nucleotide polymorphisms (SNPs): rs58542926 (E167K) in TM6SF2, rs505151 in PCSK9, rs320 in LPL and rs688 in LDLR were selected and genotyped via real time polymerase chain reaction followed by NGS (next-generation sequencing). The χ2 test and haplotype analysis were performed to analyse the associations between the four SNPs and AMI by comparing the different distribution using the SPSS V.22.0 software package.
RESULTS Obvious differences of alleles distribution in AMI cases and controls were observed in TM6SF2 rs58542926, LPL rs320 and LDLR rs688, as well as genotype distribution of rs320 and rs688 in recessive model from analysis of single gene locus. It was also found significant difference of triglyceride (TG) of LPL rs320 and APOB/APOA1 in TM6SF2 rs58542926 of total study population. In binary logistic regression analysis, TM6SF2 rs58542926 was associated with reducing acute myocardial infarction risk (TT/CC, OR=0.098, 95% CI=0.024–0.392). Stratified association analysis revealed PCSK9 rs505151 was associated with acute myocardial infarction in male group (GA/AA, OR=2.445, 95% CI=1.094–5.465) and in non-hypertension group (GA/AA, OR=2.368, 95% CI=1.013–5.533).
CONCLUSIONS The genotype TT of TM6SF2 rs58542926 was associated with reducing acute myocardial infarction risk.
GW31-e1054
Chao Sun, Baojian Zhang, Na Liu, Biao Li, Qiming Liu
Department of Cardiovascular Medicine, The Second Xiangya Hospital, Central South University
OBJECTIVES To explore the predictive values of simplified acute physiology score II (SAPS II) and Oxford acute severity of illness score (OASIS) for prognosis in patients with acute myocardial infarction (AMI).
METHODS Data including baseline information, vital signs, and some laboratory test results of adult AMI patients was extracted from medical information mart for intensive care III (MIMIC-III). SAPS II and OASIS of each patient were calculated according to the requirements of each scoring system. The patients were divided into two groups according to the occurrence of in-hospital mortality. We compared the difference of baseline information between two groups. Logistic regression analysis and receiver operating characteristic (ROC) analysis was performed to analyze the association between scoring systems and in-hospital mortality. The discharged patients were divided into four groups according to the quartile values of the score, respectively. The association between scoring systems and 1-year mortality after discharge were evaluated by Cox proportional hazards model.
RESULTS A total of 5031 adult AMI patients with in-hospital mortality rate of 14.5% (728/5031) were included finally, of which female patients accounted for 38.9% (1959/5031). When compared with the survivors, non-survivors had higher SAPS II and OASIS scores and the differences were statistically significant (P<0.001, P<0.001). Logistic regression analysis showed that both SAPS II (OR=1.043, 95% CI 1.033–1.054, P<0.001) and OASIS (OR=1.046, 95% CI 1.030–1.063, P<0.001) were independent predictors of in-hospital mortality in AMI. ROC analysis showed that the area under curve (AUC) of SAPS II and OASIS was 0.803 (95% CI 0.786–0.820, P<0.001) and 0.770 (95% CI 0.750–0.789, P<0.001), respectively. The optimal SAPS II threshold was 42 and yielded a 72.9% sensitivity and 74.4% specificity. The optimal OASIS threshold was 36 and yielded a 67.5% sensitivity and 75.1% specificity. The Hanley-McNeil test showed a significant difference of AUC between the two scoring systems (Z=4.393, P<0.001). Cox proportional hazards model showed that increased SAPS II was the independent risk factor of 1-year mortality after discharge in AMI (HR=1.017, 95% CI 1.006–1.027, P=0.002) and OASIS was not the independent predictor (HR=1.007, 95% CI 0.993–1.022, P=0.337).
CONCLUSIONS Both SAPS II and OASIS are independent predictors of in-hospital mortality in AMI. They can predict in-hospital mortality of AMI effectively, and the predictive value of SAPS II is superior to OASIS. SAPS II is also the independent predictor of 1-year mortality after discharge in AMI.
GW31-e1072
Zhichao Wang, Jun Tao
Department of Hypertension and Vascular Disease, The First Affiliated Hospital, Sun Yat-Sen University
OBJECTIVES It is well known that there is a clear correlation between neutrophil-to-lymphocyte ratio (NLR) and the occurrence of in-stent restenosis (ISR). However, few studies focused on patients with type 2 diabetes mellitus (T2DM) and evaluated the dynamic changes of NLR value to predict ISR. We aimed to assessed the predictive value of two pre-procedural NLR on ISR in patients with T2DM.
METHODS This retrospective study consecutively enrolled 96 patients with T2DM who underwent Drug-eluting stents (DES) implantation and follow-up coronary angiography (CAG). Pre-procedural blood cell parameters were analyzed in the first percutaneous coronary intervention (PCI) and the second CAG. Multivariate logistic regression analysis to determine the predictors of ISR. Receiver operating characteristic (ROC) curves assessed predictive value of NLR in ISR patients with T2DM.
RESULTS Patients were divided into 2 groups at mean follow-up of 11.0±2.2 months: Non-ISR group (n=64), ISR group (n=32). The two pre-procedural NLR levels in the ISR group were significantly higher than that in the Non-ISR group (P≤0.001), respectively. Multivariate logistic regression analysis indicated that pre-procedural NLR was an independent risk predictor for ISR in patients with T2DM (NLR1, odds ratio [OR]=1.94, 95% confidence interval [CI]: 1.08–3.50, P=0.03; NLR2, OR=3.90, 95% CI: 1.78–8.55, P=0.001). Receiver operating characteristic (ROC) curves showed that the first pre-procedural NLR (NLR1) cutoff value for predicting ISR rate was 2.86 with a sensitivity of 59% and a specificity of 73%. The second pre-procedural NLR (NLR2) cutoff value for predicting ISR rate was 2.51 with 75% sensitivity and 70% specificity.
CONCLUSIONS NLR was an independent risk factor for DES-ISR in patients with T2DM, and NLR before second CAG may have a better predictive value in identifying ISR.
GW31-e1088
Xiuxiu Xu1,2,3, Junmei Zhang2,3, Ru San Tan2,3, Ping Chai4, Qinghua Wu1, Huan Han2, Lynette L.S. Teo4, Ris Low2, Jiang Ming Fam2, Chee Yang Chin2, Mark Chan4, Adrian F. Low4, Swee Yaw Tan2,3, Terrance Chua2,3, Soo Teik Lim2,3, Liang Zhong2,3
1Department of Cardiovascular Medicine, the Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, China
2National Heart Centre Singapore, 5 Hospital Drive, 169609 Singapore
3Duke-NUS Medical School, 8 College Rd, 169857 Singapore
4National University Hospital Singapore, 1E Kent Ridge Road, 119228 Singapore
OBJECTIVES Fractional flow reserve (FFR), the ratio of mean blood pressure distal to the coronary artery (CA) stenoses to mean aortic blood pressure measured under drug-induced hyperemia during invasive coronary angiography (ICA), is the gold standard for diagnosing CA ischemia. We studied the effects of gender and CA severity on FFR measurements.
METHODS Subjects underwent computed tomographic coronary angiographic (CTCA) and clinically indicated ICA with FFR measurements. CA lesions were stratified according to ICA-assessed diameter stenosis (DS) using quantitative coronary analysis: <50% (mild), 50–69% (moderate), 70–99% (severe).
RESULTS One hundred and nine subjects (mean age 59.9±9.2 years, 32 females) from multi-centers were enrolled and FFR measurements were performed in 169 CA stenoses. FFR was similar between females and males in mild stenosis (0.90±0.04 vs. 0.90±0.06, P=0.70) and moderate stenoses (0.83±0.09 vs. 0.85±0.06, P=0.40), but were significantly different in severe stenoses (0.77±0.09 vs. 0.69±0.16, P=0.02). FFR was not influenced by the location of the stenoses (left artery descending, circumflex, and right coronary artery) (all P>0.05) between genders.
CONCLUSIONS There were significant gender differences in FFR measured in severe but not mild and moderate CA stenoses. The mechanism of this observation is unclear and how this will affect the physiological assessment of CA stenosis warrants further studies.
GW31-e1131
Huiwen Zhang, Yuanlin Guo, Naqiong Wu, Ying Gao, Ruixia Xu, Qian Dong, Jing Sun, Jianjun Li
Fu Wai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College
OBJECTIVES Heart-type fatty acid binding protein (H-FABP), a cardiac biomarker, which has been used as a diagnostic marker of myocardial injury and a predictive indicator for clinical outcomes in patients with acute coronary syndrome. However, its prognostic utility in patients with premature coronary artery disease (PCAD) has been less investigated. The aim of this study was to assess the relationship between H-FABP and cardiovascular events (CVE) in patients with PCAD.
METHODS A total of 2828 consecutive patients with PCAD (defined as male <55 years, female <65 years) were enrolled and followed-up for CVEs. The CVEs were defined as cardiovascular death, myocardial infarction, stroke and coronary revascularization. Plasma H-FABP levels were measured using the Latex immunoturbidimetric method. The association between H-FABP with CVEs was analyzed by Cox regression analysis and Kaplan-Meier analysis.
RESULTS There were 205 CVEs occurred duringa median of 49 months follow-up. H-FABP levels were significantly higher in patients with CVEs compared with those without CVEs (mean: 2.71 ng/mL vs. 2.15 ng/mL, P<0.001). In addition, patients with the highest level of H-FABP had increased rate of CVE compared with ones in the lowest groups (P<0.05). Moreover, in Cox regression analysis, data indicated that elevated H-FABP levels were independently correlated with a higher risk of CVEs (P<0.05) after adjusted for confounders.
CONCLUSIONS Elevated levels of H-FABP were independently associated with increased risk of CVEs in patients with PCAD. These data might provide novel information of H-FABP in predicting adverse outcomes in patients with PCAD.
GW31-e1140
Ting Ting Wu, Xiang Xie, Yi Tong Ma
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830054, P.R. China
OBJECTIVES Prognostic stratification of patients with coronary artery disease (CAD) may improve the clinical management and facilitate clinical trials. We aimed at developing a scoring system capable of providing prognostic stratification of patients with CAD.
METHODS A retrospective study (identifier: ChiCTR-INR-16010153) contains 6046 CAD patients were evaluated finally. Using the Kaplan-Meier analysis with the cut off value of 7.985, patients were divided into two groups (ABIC score ≥7.985, n=2808; <7.985, n=3238). The primary outcome long-term mortality and secondary endpoints mainly major adverse cardiovascular and cerebrovascular events (MACCEs) were recorded. Multivariate Cox regression models were used to determine risk factors for mortality and MACCEs.
RESULTS A total of 309 patients died during the following up time. There were significantly higher adverse events in the high group compared to that in the low group with 192 (6.8%) vs. 117 (3.6%), cardiac death 148 (5.3%) vs. 103 (3.2%) and MACCE 422 (15.0%) vs. 440 (13.6%) respectively. However, the frequency of MACCE did not differ significantly between the two groups. After adjusting for eight confounders, the multivariate Cox proportional hazards model showed that when ABIC score ≥7.985, the incidence of all cause mortality would be increased 1.7 times (adjusted HR=1.729 (1.347–2.218), P<0.001), and 1.5 times in cardiac death (adjusted HR=1.482 (1.126–1.951), P=0.005).
CONCLUSIONS The present study indicated that ABIC score predicts high long-term mortality risk for PCI patients, ABIC score might be a potential prognostic model for patients with CAD.
GW31-e1142
Ting Ting Wu, Xiang Xie, Yi Tong Ma
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, P.R. China
OBJECTIVES To investigate the long-term prognosis of average platelet volume (MPV) to platelet count (PC) ratio in acute coronary syndrome (ACS) patients underwent percutaneous coronary intervention (PCI).
METHODS A retrospective cohort study including 2751 ACS patients after PCI in the First Affiliated Hospital of Xinjiang Medical University from January 2008 to December 2016 was conducted. All laboratory data including platelet parameters, biochemical data, and cardiovascular disease-related risk factors were collected from the medical records. Follow-up endpoints were all-cause mortality, cardiac deaths, major cardiovascular adverse events (MACE), stroke, and bleeding events. Follow-up data was obtained from telephone interview with the patients or family members.
RESULTS The optimal clinical cut off point of MPV/PC value of patients was 0.043 selected by ROC curve. Baseline data analysis showed that compared with the low MPV/PC group (MPV/PC ≤0.043), cardiac deaths 54 (3.1%) vs. 62 (6.3%), all-cause mortality 73 (4.1%) vs. 71 (7.2%), and MACE events 424 (24.0%) vs. 274 (27.9%) were significantly higher in the high-value MPV/PC group. Multivariate Cox regression analysis showed that all cause mortality increased by 1.7 times in the MPV/PC high-value group (OR 1.713, 95% CI 1.225–2.397, P=0.002); the cardiac deaths will increased by 2 times (OR 2.037, 95% CI 1.400–12.963 P<0.001).
CONCLUSIONS High MPV/PC ratio is associated with poor prognosis after PCI and it could be an independent risk factor for all-cause mortality and cardiac deaths.
Keywords : Acute coronary syndrome, MPV, PLT, prognosis
GW31-e1167
Jing Liu1,2, Daomin Yao2, liang Xie1, Yanming Liu1, Jianbin Gong1
1Nanjing General Hospital of Nanjing Military Command
2Nanjing University Of Chinese Medicine
OBJECTIVES To identify differentially expressed microRNAs (miRNAs) in rat myocardial infarcted tissues and predict their interaction with lncRNAs and target genes, as well as explore potential pathophysiology mechanisms in myocardial infarction.
METHODS A rat model of myocardial infarction was established by ligating the left anterior descending coronary artery. Trizol was used to extract total RNA from infarcted myocardial area for Microarray detection. Bioinformatics methods were used to predict interaction lncRNAs, target genes prediction, and functional enrichment of miRNAs that were significantly differently expressed. Find possible lncRNA-miRNA-mRNA regulatory networks finally.
RESULTS The elevation of ST segment of ECG showed that the rat model of myocardial infarction was successfully prepared. Microarray results showed that there were 19 significantly differently expressed miRNAs. Eight of these miRNAs (miR-21, miR-132, miR-222, miR-223-3p, miR-146a/b, miR-181b, miR-449a-5p, miR-122) have been proven to be myocardial infarction treatment Candidates. Whether seven miRNAs (miR-365-5p, miR-490-5p, miR-6333, miR-30c-1-3p, miR-3591, miR-3596c, miR-877) are related to myocardial infarction has not been confirmed. There may be several new lncRNA-miRNA-mRNA mechanisms in the development of myocardial infarction. ENSRNOT00000076620-miR-146b-5p-STAT3/Rnf7/Qrsl1 may be involved in the process of cardiomyocyte apoptosis and mitochondrial damage during myocardial infarction. ENSRNOT00000071991-miR-122-Deptor may inhibit the autophagy of cardiomyocytes and exacerbate myocardial infarction
CONCLUSIONS The ternary relationship of lncRNA-miRNA-mRNA obtained in this study may provide possible research directions and a certain theoretical basis for further exploration of the molecular level pathological mechanism of myocardial infarction, as well as finding new therapeutic targets for myocardial infarction.
GW31-e1168
Qiuzhen Lin1,2,3,4, Qiming Liu1,2,3,4
1Department of Cardiovascular Medicine, The Second Xiangya Hospital, Central South University
2Modern Cardiovascular Disease Clinical Technology Research Center of Hunan Province
3Cardiovascular Disease Research Center of Hunan Province
4Research Institute of Blood Lipid and Atherosclerosis, Central South University
OBJECTIVES It was recognized that malnutrition and an increased risk of adverse outcomes in patients with different cardiovascular diseases had a close relationship. The aim of this study was to use a novel Japanese calculated nutritional index in Chinese patients with coronary heart disease (CHD).
METHODS This study was a retrospective observational analysis. There was 11,935 patients with CHD at department of cardiovascular medicine, the Second Xiangya Hospital, Central South University from 2011 to 2019 were recruited in this study. The novel Japanese nutritional index was calculated by the formula: Triglycerides (TG)×Total Cholesterol (TC) ×Body Weight (BW) Index (TCBI)=TG×TC×BW/1000 (TG and TC: mg/dL, and BW: kg). The most often used conventional nutritional index, Geriatric Nutritional Risk Index (GNRI)=14.89×serum Alb (g/dL)+41.7×(measured BW (kg)/ideal BW (kg)). Ideal BW was calculated using Lorentz-formula. Ideal BW=(height (cm)-100)-(height (cm)-150)/4 for men and (height (cm)-100)-(height (cm)-150)/2 for women. Then, we calculated the values of TCBI and GRNI of the included patients. Because the normality test indicated that both of them were non-naormal distribution, we used the Spearman non-parametric correlation coefficient between TCBI and GNRI to evaluate the predictive ability of the novel nutritional index of Chinese patients with CHD.
RESULTS All subjects were 22~94 years old, including 3406 (28.5%) women and 8529 (71.5%) men. The BMI ranged from 15.06 kg/m2 to 40.09 kg/m2. The means of TG and TC were 153.17 mg/dL and 150.31 mg/dL, respectively. The Spearmen non-parametric correlation coefficient between TCBI and GNRI was 0.397, which was similar to the result of the study committed in Japan, indicating modest correlation.
CONCLUSIONS This study indicated that TCBI may be a useful prognostic indicator in Chinese patients with CHD.
GW31-e1174
Yulan Ma, Youlu Shen, Lin Li, Shiming Fan, Xiuying Chai, Huizhen Tian, Yue Lin, Zhongshan Gao, Ming Ren
Department of cardiology Afflicted hospital of Qinghai University
OBJECTIVES This study was designed to demonstrate the structural characteristics of gut microbiota of Tibetan patients with coronary artery disease in the Qinghai-Tibet Plateau, and to preliminarily determine the relationship between it and the pathophysiology of coronary artery disease in the Tibetan population of the Qinghai-Tibet Plateau.
METHODS The fecal tissues of patients with coronary artery disease and healthy Tibetans living on the Qinghai-Tibet Plateau, and those Han people with coronary artery disease living in Xining and Wuhan were collected. The 16S rRNA of the gut microbiota of the above specimens was sequenced and analyzed by bioinformatics.
RESULTS (1) There are certain differences in the α diversity and β diversity of the gut microbiota between high-altitude Tibetan patients with coronary artery disease and healthy individuals, and high-altitude Tibetan patients with coronary artery disease showed higher proportion of the beneficial bacteria Verrucomicrobia and its main member Akkermansia. (2) Compared with Xining and Wuhan Han patients with coronary artery disease, the α diversity of the gut microbiota in high-altitude Tibetan patients with coronary heart disease was significantly increased, and the β diversity also formed different cluster. The proportion of pathogenic bacteria Streptococcus, Escherichia/Shigella, and Klebsiella decreased. While the proportion of beneficial bacteria Faecalibacterium, Prevotella, Catenibacterium, and Lactobacillus increased.
CONCLUSIONS These results indicated that the gut microbiota of high-altitude Tibetan patients with coronary artery disease presented protective features, which might inhibit the progression of coronary artery disease.
GW31-e1177
Ting Ting Wu, Wan Rong Wang, Xiang Xie, Yi Tong Ma
Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, P.R. China
OBJECTIVES Previous studies have linked HDL cholesterol (HDL-C) to cardiovascular protection, and the general public refers to it colloquially as good protein. In order to achieve therapeutic and health benefits, some drugs are used to specifically increase HDL-C levels. However, some recent studies have suggested that higher HDL-C is not better, and that there is a u-shaped curve between HDL-C levels and coronary artery disease (CAD). Therefore, the purpose of this study is to further clarify the role of HDL-C by analyzing the fractions of HDL-C.
METHODS We adopted the extreme study strategy to conduct subgroup analysis of HDL-C fractions in 17 patients with CAD whose HDL-C content were >50 mg/dL and 14 healthy controls whose HDL-C content were <30 mg/dL, so as to clarify the correlation between HDL-C fractions and CAD. From January to March 2019, 144 subjects in the case group and control group with HDL-C content of 40–60 mg/dL were admitted in the first affiliated hospital of Xinjiang medical university to further verify the results of the extreme value strategy study.
RESULTS The patients fasting serum lipid parameters (total cholesterol, triglycerides, low-density lipoprotein cholesterol, HDL-C, and cholesterol in 10 lipoprotein fractions were determined. For the comparison of the extreme set, there were no significant differences in small HDL-C fractions between groups. Decreases in large and intermediate HDL-C fractions 1–7 were observed in CAD group. In the verified group, intermediate HDL-C fractions of 4 and 7 were decreased significantly. In CAD group, decreases of intermediate and large HDL-C fractions were noticed, and the reverse in health group. After adjusted for other CAD risk factors, large and intermediate HDL-C fractions were identified to be independent protect factors for CAD by multivariate logistic analysis. When large and intermediate HDL-C fractions increase 1 unit, the incidence of CAD would be decreased 0.891 times (adjusted OR=0.891 (0.823–0.966), P=0.005), and 0.808 times (adjusted OR=(0.726–0.898), P<0.001) respectively. When intermediate HDL-C fractions ≥27.5 mg/dL, the risk will decrease to 0.205 (adjusted OR=0.205 (0.070–0.604).
CONCLUSIONS The results show that large and intermediate HDL-C fractions were identified to be an independent protect factor for CAD, and that the intermediate HDL-C fractions is a more accurate indicator of HDL-C function than existing indices.
GW31-e1198
Yue Liu, Sida Jia, Deshan Yuan, Na Xu, Lin Jiang, Lianjun Xu, Ying Song, Zhan Gao, Jue Chen, Yuejin Yang, Runlin Gao, Bo Xu, Jinqing Yuan
Fuwai Hospital, Chinese Academy of Medical Sciences
OBJECTIVES The relationship between inflammatory markers, high-sensitivity C-reactive protein-to-albumin ratio (CAR) and fibrinogen-to-albumin ratio (FAR), and coronary artery disease has not been fully evaluated. This study aimed to investigate the predictive value of admission CAR and FAR for 5-year outcomes in patients undergoing percutaneous coronary intervention.
METHODS Data came from a large prospective cohort study with 10,724 patients consecutively enrolled between January 2013 and December 2013. The primary endpoint was all-cause death. And the secondary endpoints were cardiac death and major cardiovascular and cerebrovascular events (MACCEs). The optimal cutoff value of each ratio for all-cause mortality was determined according to the receiver operating curve.
RESULTS A total of 6617 patients were eligible for analysis. Patients with high CAR (>0.035) or FAR (>0.075) presented a high-risk profile for complications. During 5-year follow-up, significantly more all-cause death [CAR: 2.7 versus (vs) 4.9%; FAR: 2.6 vs. 5.0%, both P<0.001) and cardiac death (CAR/FAR: 1.4 vs. 3.1%, P<0.001) occurred in both high ratio groups, while MACCEs were only different between low and high FAR groups (21.8 vs. 24.9%, P=0.006). After adjusting confounders, both ratios remained independently predictive of all-cause death [CAR: hazard ratio (HR) 1.65 (1.16–2.35), P=0.005; FAR: HR 1.44 (1.01–2.05), P=0.046] and cardiac death [CAR: hazard ratio (HR) 2.20 (1.35–3.57), P=0.001; FAR: HR 2.04 (1.23–3.36), P=0.005]. The associations were also relatively consistent in subgroups with no significant interactions. Additionally, incorporating CAR or FAR further improved the reclassification ability of SYNTAX score II for mortality prediction.
CONCLUSIONS CAR and FAR were independent predictors for long-term prognoses after percutaneous coronary intervention. The inflammatory biomarkers could enhance the predictivity of SYNTAX score II for mortality.
GW31-e1210
Ling Xu, Wei Zhao, Cui Ming
Department of Cardiology, Peking University Third Hospital, NHC Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides. Beijing 100191, China
OBJECTIVES To prove the clinical efficacy of enhanced external counterpulsation (EECP) in patients with coronary heart disease after incomplete revascularization.
METHODS Forty subjects with coronary heart disease after incomplete revascularization were assigned to EECP group (standard course of EECP combined with drug therapy) and control group (drug therapy) according to their treatment intention, with 20 patients in each group. All subjects underwent three-dimension speckle tracking endocardiography (3D-STE), transthoracic echocardiography (TTE) examinations at baseline, 18 hours of treatment (control group for 3–4 weeks of follow-up), 35 hours of treatment (control group for 7 weeks of follow-up).
RESULTS After 35 hours of EECP treatment, the myocardial strain parameters Twist (7.4±4.9° vs. 2.8±3.5°, P=0.016) and Torsion (1.5±0.7°/cm vs. 0.8±0.6°/cm, P=0.006) improved significantly, while the 3D-STE and TTE related parameters in the control group did not change significantly during the follow-up period. Correlation analysis found that in the control group the improvement of Twist was negatively correlated with the baseline coronary Gensini score (r=-0.597, P=0.015), which was not observed in the EECP group. In the subgroup analysis, it was found that in the patients without diabetes mellitus (DM), EECP had a more significant effect on the improvement of myocardial strain parameters than the patients with DM.
CONCLUSIONS For patients with coronary heart disease after incomplete revascularization, the standard course of EECP treatment can increase myocardial strain parameters Twist and Torsion, suggesting that EECP can effectively improve cardiac systolic function in such patients.
GW31-e1225
Zhijie Mao1, Ya Lin1, KangWei Wang1, JianJian Huang2, LuYa Wang1, YiHe Chen1, WeiJian Huang1, Zhouqing Huang1
1The Key Laboratory of Cardiovascular Disease of Wenzhou, Department of Cardiology, The First Affiliated Hospital of WenZhou Medical University, WenZhou, ZheJiang, China
2Department of Anaesthesiology, Wenzhou Medical University, Wenzhou, Zhejiang, China
OBJECTIVES Acute kidney injury (AKI) is a serious and common complication of acute myocardial infarction (AMI), which significantly increases mortality. However, the definition of acute kidney injury (AKI) based on urine output is usually neglected in the prognosis of patients with Acute Myocardial Infarction. This study is to access the influence of three AKI definitions, especially based on urine output, on the accuracy of mortality prediction in patients with AMI.
METHODS This study was a retrospective study based on the open MIMIC-III database. We enrolled 1953 patients with AMI. Exposure is based on the maximum AKI stage after admission to the ICU. Multiple Cox proportional hazards models were used to test the association between different definitions of acute kidney injury and all-cause mortality. The discriminant degree of the model was compared using the area under the ROC curve (AUC) and integrated discrimination improvement (IDI). In addition, we do a nomogram according to AKI-UO to predict in-hospital mortality of patients with AMI. The predictive accuracy of the nomogram was according to the c-index and calibration using Hosmer-Lemeshow (H-L) chi-square statistics.
RESULTS Of the 1953 participants, the mean (SD) age was 67.4 (14.1) years; the mean weight was 81.2 (9.7) kg; and the serum creatinine (scr) initial was 1.2±0.8 mg/dL. Overall, 1264 (64.7%) study participants were male and 118 (6.0%) have old myocardial infarction (OMI), 1548 (79.3%) have coronary diseases, 944 (48.3%) had hypertension, 518 (26.5%) had diabetes mellitus. 191 (9.8%) patients appear in-hospital death. After a total of 90 days of follow-up, 293 (15.0%) participants death. AKI as defined by AKI-uo, AKI-scr, AKI-kdigo occurred in 54, 38, 65.6% of the study patients during hospitalization, respectively. And AKI-UO, AKI-Scr, and AKI-KDIGO were independent predictors of in-hospital mortality (with adjusted OR of 3.4 (95% CI, 2.1–5.7), 2.4 (95% CI, 1.7−3.5) and 3.7 (95% CI, 1.9−7.2), respectively). AUC was 0.8238 for the clinical model containing age (per 10 years increase), heart rate, surgery, PT, haemoglobin-min and glucose-max. Meanwhile, the addition of AKI-UO, AKI-Scr, AKI-KDIGO to the clinical model improved the in-hospital mortality models. (AUC plus AKI-UO: 0.8625, plus AKI-Scr: 0.8537, and plus AKI-KDIGO: 0.8612; integrated discrimination improvement (IDI) was 5.9% (P<0.0001), 1.6% (P=0.0038), and 1.3% (P=0.0002), respectively). With regard to 90-day mortality, the IDI of AKI-UO, AKI-Scr, and AKI-KDIGO was 6.2% (P<0.0001), 2.3% (P<0.0001), and 2.1% (P<0.0001), respectively. Finally, in our study, we performed a nomogram to predict in-hospital mortality of patients with AMI based on AKI-UO. The nomogram had a c-index of 0.8625 (95% CI 0.8386–0.8864) and a cut-off value of −2.4735, with a sensitivity of 87% and a specificity of 70%. The Hosmer-Lemeshow chi-square test indicated that the nomogram was well-calibrated (chi-square: 9.182, P=0.327).
CONCLUSIONS AKI-UO is an independent predictor of all-cause mortality in AMI patients. The addition of AKI-UO to the traditional clinical model significantly improved its predictive accuracy for in-hospital and 90-day all-cause mortality.
GW31-e1238
Wencai Zhang, Zhanying Han
The First Affiliated Hospital of Zhengzhou University
OBJECTIVES The aim of this study was to explore the mechanisms accounting for the expansion of myeloid-derived suppressor cells (MDSCs) in patients with acute coronary syndrome (ACS).
METHODS The frequencies of circulating CD14+HLA-DR-/low MDSCs were detected by flow cytometry, and miR-21-5p expression was analyzed by real-time reverse transcription polymerase chain reaction (RT-PCR). Correlation analyses between MDSC frequencies and miR-21 levels were also performed. Then, to determine the role of miR-21 in the expansion of MDSCs, cells were transfected with miR-21-m, miR-NS-m or control. In addition, levels of phosphatase and tensin homolog (PTEN) and phospho-signal transducer and activator of transcription-3 (p-STAT-3) were detected by western blotting.
RESULTS The frequencies of circulating CD14+HLA-DR-/low MDSCs and the levels of miR-21-5p were both obviously increased in patients with ACS, and there was a positive correlation between these parameters. Furthermore, treatment with miR-21-m markedly increased miR-21 expression and thus promoted the expansion of MDSCs. Meanwhile, treatment with miR-21-m markedly decreased PTEN expression and increased STAT-3 activity. In addition, disruption of STAT-3 activity with the specific inhibitor S3I201 nearly abolished miR-21-induced MDSC expansion.
CONCLUSIONS Our results demonstrated that miR-21 contributed to the expansion of MDSCs through reduction of PTEN expression, thereby leading to STAT-3 activity, in patients with ACS.
GW31-e1252
Haobo Teng, Chao Guo, Shubin Qiao
Center of Coronary Heart Disease, Fuwai Hospital, National Center for Cardiovascular Diseases
OBJECTIVES To investigate the impact of shock index before IABP insertion on recent prognosis of patients with cardiogenic shock complicated by acute myocardial infarction supported by intra-aortic balloon pump.
METHODS A total of 103 patients diagnosed as cardiogenic shock complicated by acute myocardial infarction admitted in our hospital from 2014.6 to 2018.5 were enrolled in our study. We collected the data from medical records and investigated their clinical manifestation and laboratory examination, as well as 28-day mortality retrospectively, we calculated the shock index (ratio of heart rate to systolic blood pressure) before IABP insertion.
RESULTS Compared with lower SI before IABP insertion, patients with higher SI had higher proportion of anterior infarction (81.5 vs. 61.2%, P=0.022), history of PCI (24.1 vs. 8.16%, P=0.030), left main as culprit vessel (31.5 vs. 12.2%, P=0.019), and final TIMI flow ≤2 (55.5 vs. 26.5%, P=0.003), invasive ventilation (40.7 vs. 20.4%, P=0.026) as well as 28-day-mortality (81.5 vs. 61.2%, P=0.022). SI could predict the recent prognosis, with a cut-off of 1.625, a sensitivity of 0.655 and a specificity of 0.708, AUC of ROC was 0.713; On multiple analysis, SI, together with final TIMI flow, arterial pH and creatinine were independent predictive factors of recent prognosis in this population.
CONCLUSIONS Among patients with cardiogenic shock complicated by acute myocardial infarction, higher SI before IABP insertion was associated with poorer prognosis, SI was an independent risk factor of recent mortality, and could predict the recent prognosis well.
GW31-e1304
Haoyu Wu, Lijun Wang, Chen Wang, Shanshan Gao, Zuyi Yuan
Department of Cardiovascular Medicine, First Affiliated Hospital of Xi’an Jiaotong University
OBJECTIVES Atherosclerosis (AS) is the pathophysiological basis of coronary artery disease (CAD) with inflammation and cholesterol metabolism disorder as its two basic hallmarks. Monocytes in the circulation are the main cells secreting various pro-inflammatory factors, and play an extremely important role in the initiation of coronary AS, plaque formation and stability. Studies have shown that vulnerable plaques exhibit significant monocyte infiltration, and monocytes in the peripheral blood of patients are mostly activated. High-density lipoprotein cholesterol (HDL-C) is a lipid component that exerts anti-AS effects, by promoting cholesterol efflux and preventing monocyte activation, proliferation and adhesion to endothelium. Increased monocyte counts and decreased HDL-C levels have been reported to be associated with inflammatory disorders. Recently, monocyte to HDL-C ratio (MHR) has emerged as a novel and useful marker in different disease models, such as CAD, diabetes mellitus (DM), saphenous vein graft disease (SVGD) and acute ischemic stroke (AIS). In a cross-sectional study on 1229 CAD patients, Akboga et al. found that MHR is significantly correlated with the burden of AS. Another study also found that MHR was obviously higher in 428 stable CAD patients with SYNTAX score of ≥23 than those with SYNTAX score of <23. However, the abovementioned studies usually had relatively small sample size, and few study has ever investigated the association of MHR with the severity of CAD assessed by Gensini score in a large scale population. Therefore, in this study, we evaluated the association of MHR with the severity and complexity of CAD using the Genisi score and determined the ability of MHR in predicting severe CAD and acute atherothrombosis events.
METHODS A total of 4950 patients who presented to the Department of Cardiovascular Medicine of the First Affiliated Hospital of Xi’an Jiao University for angiography between 2017 January and 2018 July were recruited. The total population included 3930 CAD patients and 1020 non-CAD patients. The CAD patients were classified into four groups according to the quartile of the MHR (≤0.28, N=1218; 0.28–0.39, N=1262; 0.39–0.53, N=1209; >0.53, N=1261). CAD severity was quantified according to the Gensini score. A receiver operating characteristic (ROC) curve analysis was also performed to predict severe CAD and acute coronary thrombotic events.
RESULTS MHR was significantly higher in the CAD group than in the non-CAD group (0.45±0.22 vs. 0.35±0.17, P<0.001) and had a significant positive correlation with Gensini score. Compared with lower MHR value, a MHR in the fourth quartile was strongly associated with severe CAD and acute coronary thrombotic event after adjusting for baseline factors. Receiver-operating characteristic (ROC) curve analysis showed that combination of MHR and traditional risk predictors could better predict severe CAD especially acute coronary thrombosis events such as non-ST-elevation myocardial infarction (NSTEMI) and acute ST-elevation myocardial infarction (ASTEMI).
CONCLUSIONS We demonstrated in this study that MHR is positively associated with the severity of CAD and MHR is an independent indicator to predict severe CAD and acute coronary thrombosis events.
GW31-e1333
Xuyang Wang, Xiaogang Guo
Department of Cardiology, The First Affiliated Hospital, Zhejiang University School of Medicine, 79 Qingchun Road
OBJECTIVES The aim of this study is to evaluate the effect of nicorandil on IMR in IHD patients with PCI and other intervention therapy by means of a meta-analysis.
METHODS CENTRAL, PUBMED, EMBASE, CNKI, and Wanfang electronic databases were searched for randomized controlled trials and non-randomized controlled trials published until October 2018. We included clinical controlled trials comparing the effect of nicorandil on IMR versus placebo or other pharmacological interventions in IHD patients with PCI and other intervention therapy.
RESULTS Four trials including a total number of 232 participants were identified. The baseline IMR between the nicorandil group and the control group showed no statistically difference; the posttreatment IMR between the nicorandil group and the control group showed significant statistically difference; the change of IMR in the nicorandil group was significant larger than that in the control group.
CONCLUSIONS Nicorandil could reduce the value of index of microcirculatory resistance (IMR) in the IHD patients who has received interventional treatment, sequentially improve coronary microcirculation function of patients.
GW31-e1335
Guang Yang, Chunsheng Lin
Shaanxi Provincial People’s Hospital, Department Cardiology
OBJECTIVES Myocardial infarction (MI) was a severe cardiovascular disease resulted from acute, persistent hypoxia or ischemia condition. Additionally, MI generally led to heart failure, even sudden death. A multitude of researches proposed that long non-coding RNAs (lncRNAs) frequently participated in the regulation of heart diseases. The specific function and molecular mechanism of SOX2-OT in MI remained unclear. The current research was aimed to explore the role of SOX2-OT in MI.
METHODS Bioinformatics analysis (DIANA tools and Targetscan) and a wide range of experiments (CCK-8, flow cytometry, RT-qPCR, luciferase reporter, RIP, caspase-3 activity, trans-well and western blot assays) were adopted investigate the function and mechanism of SOX2-OT.
RESULTS We discovered that hypoxia treatment decreased cell viability but increases cell apoptosis. Besides, lncRNA SOX2-OT expression was up-regulated in hypoxic HCMs. Hereafter, we confirmed that SOX2-OT could negatively regulate miR-27a-3p levels by directly binding with miR-27a-3p, and miR-27a-3p also could negatively regulate SOX2-OT levels. Furthermore, knock-down of SOX2-OT promoted cell proliferation, migration and invasion, but limited cell apoptosis. However, these effects were reversed by anti-miR-27a-5p. Besides, we verified that miR-27a-3p binding with the 3'UTR of TGFBR1 and SOX2-OT regulated TGFβR1 level by collaborating with miR-27a-3p in HCMs. Eventually, rescue assays validated that the influence of SOX2-OT silence or miR-27a-3p overexpression on cellular processes in cardiomyocytes injury was counteracted by TGFBR1 overexpression.
CONCLUSIONS Long non-coding RNA SOX2-OT exacerbated hypoxia-induced cardiomyocytes injury by regulating miR-27a-3p/TGFβR1 axis, which may provide a novel insight for heart failure treatment.
GW31-e1362
Lu Lin1, Jianjian Huang2, Xue Xia1, Zhongqiu Lu1, Bozhi Ye1, Zhouqing Huang1
1The Key Laboratory of Cardiovascular Disease of Wenzhou, Department of Cardiology, The First Affiliated Hospital of WenZhou Medical University, WenZhou, ZheJiang, China
2Department of Anesthesiology, Wenzhou Medical University
OBJECTIVES At present the pathogenesis of atherosclerosis has not been fully elucidated which including the lipid infiltration oxidative stress Ca2+ overload abnormal immune function and so on While as a theory of relatively new inflammatory mechanism attracts much attention in recent years Studies have shown that the inflammatory response mediated by mononuclear macrophage and foam cells plays a key role in the pathogenesis of atherosclerosis MMPs and EMMPRIN overproduced and released by mononuclear macrophage and foam cells result in the rupture of atherosclerotic plaque through degrading the extracellular Moreover as another receptor protein high expressed in mononuclear macrophage and foam cells P2X7R also regulate the progress of atherosclerosis Recent studies show that berberine as a traditional Chinese medicine could reduce the risk factors of atherosclerosis such as blood glucose and blood fat which add a new entry point to treat the inflammation mediated by mononuclear macrophage and foam cells. We aimed to study whether P2X7R regulate the expression of MMP9 and EMMPRIN in oxLDL stimulated macrophages Moreover the study is to investigate whether berberine could inhibit the expression of P2X7R in mononuclear macrophage and foam cells and its potential mechanisms.
METHODS PMA differentiated macrophages were stimulated with ox-LDL for different time to detect the protein expression of P2X7R by Western blot analysis Then PMA-differentiated macrophages were pretreated with A-438079 or berberine for 1 hour before induced by ox-LDL 50 μg/mL for 48 hours Protein and total RNA were collected for Real-time PCR and Western blot analysis respectively Zymography was used to determine the MMP-9 activity through the culture supernatants.
RESULTS P2X7R was highly expressed in oxLDL stimulated macrophages in a time dependent manner The inhibition of P2X7R led to the suppression of EMMPRIN and MMP9 both at the protein and mRNA levels and that also reduced MMP9 activity Moreover P2X7R is necessary for the phosphorylation of AMPKα and MAPK pathway (ERK1/2 p38 JNK) which further regulated the expression of EMMPRIN and MMP9 We also observed that berberine suppressed the expression of P2X7R EMMPRIN and MMP9 through AMPKα and MAPK pathway in a dose dependent manner.
CONCLUSIONS In oxLDL stimulated macrophages berberine could suppress the expression of P2X7R in a dose dependent manner which further reduce the expression of EMMPRIN and MMP 9 with the participation of AMPK α and MAPK pathway.
GW31-e1368
Ting Sun, Wanlin Zhan, Lijiang Wei, Zuojun Xu, Zhaofang Yin, Li Fan, Yang Zhuo, Jingchao Hu, Changqian Wang
Department of Cardiology, Shanghai Ninth People’s Hospital, Shanghai JiaoTong University School of Medicine
OBJECTIVES ANGPTL3 and ANGPTL4 are regulators of lipid metabolism and confirmed to associate with coronary artery disease (CAD). However, whether serums of ANGPTL3 and ANGPTL4 can predict the degree of coronary stenosis is still uncertain. In this study we investigate the role of ANGPTL3 and ANGPTL4 in atherosclerosis development via a mechanism independent of its effect on plasma lipid levels.
METHODS Three hundred and five consecutive patients with coronary heart diseases referring to Departments of Cardiology of Shanghai Ninth People’s Hospital between August 2016 and December 2017 were enrolled in this retrospective study. Coronary angiography was performed by two expert cardiologists who were blinded in the blood test results. Competitive ELISA Kits for human ANGPTL3 and ANGPTL4 were used. Univariate and Multivariate logistic regression models were performed to discriminate the risk factors of atherosclerosis.
RESULTS The serum A3 levels were higher in the atherosclerosis group (coronary stenosis ≥30%) (51.71±52.67 ng/mL) than those in the coronary without obvious stenosis group (coronary stenosis <30%) (24.65±10.32 ng/mL) and ANGPTL4 levels were lower in the atherosclerosis group (561.86±716.07 ng/mL) compared with the coronary with no obvious stenosis group (626.94±729.88 ng/mL). There was a positive association between the serum levels of ANGPTL3 and the severity of coronary atherosclerosis and no significant association between ANGPTL4 levels and atherosclerosis development. In addition, the ROC curve analyses indicated that ANGPTL3 concentrations of equal to or more than 29.1 ng/mL can predict atherosclerosis (sensitivity=72.9%, specificity=73.5%) and ANGPTL4 levels of lower than 313 ng/mL was a predictor of atherosclerosis (sensitivity=71%, specificity=60%). However, in the present study, we failed to find correlations of ANGPTL3 and ANGPTL4 with clinical and biochemical parameters.
CONCLUSIONS ANGPTL3 may promote the development of atherosclerosis and increase the risk of atherosclerosis while ANGPTL4 may protect against atherosclerosis and its levels did not differ in various degree of coronary stenosis. ANGPTL3 and ANGPTL4 independently exerted an essential influence on atherosclerosis and may be better predictors of atherosclerosis than lipids.
HYPERTENSION
GW31-e0033
Sulan Huang
The First People’s Hospital of Changde
OBJECTIVES To explore the heart rate characteristics of patients with primary aldosteronism (PA) and its relationship with renin and aldosterone.
METHODS This study is a retrospective case-control study. A total of 195 cases of patients with PA diagnosed for the first time in our hospital from January 2015 to December 2019 were included in this study. Meanwhile, 195 cases of patients with primary hypertension (PH) were selected as the control group. The resting heart rate, 24 h average heart rate, day average heart rate, night average heart rate and serum potassium were observed and compared between the two groups. The relationship between renin, aldosterone and aldosterone/renin activity rates [plasma aldosterone concentration (PAC)/plasma renin activity (PRA) rate, ARR] and heart rate was studied by correlational analysis. Then the effects of renin, aldosterone and ARR on heart rate were analyzed by multiple linear regression.
RESULTS (1) The resting heart rate, 24 h average heart rate, day average heart rate and night average heart rate of PA patients were all slower than those of PH patients (P<0.05). Compared the heart rate of PA patients with different serum potassium levels, the resting heart rate, 24 h average heart rate and day average heart rate of PA patients in the hypokalemia group were slower than those in the normal potassium group (P<0.05). The resting heart rate, 24 h average heart rate, day average heart rate and night average heart rate of the adenoma group were all faster than those of the hyperplasia group, but the difference was not statistically significant (P>0.05). (2) Correlation analysis showed that renin was positively correlated with resting heart rate, 24 h average heart rate, day average heart rate and night average heart rate (P<0.05). The aldosterone/renin ratio (1nARR) was negatively correlated with resting heart rate, 24 h average heart rate, day average heart rate and night average heart rate (P<0.05). (3) Multiple linear regression analysis showed that after adjusting for age, gender and serum potassium levels, lnARR level had independent predictive value for resting heart rate, 24 h average heart rate, day average heart rate and night average heart rate (P<0.05).
CONCLUSIONS The resting heart rate, 24 h average heart rate, day average heart rate and night average heart rate of PA patients were all slower than those of PH patients. LnARR was an independent predictor of resting heart rate, 24 h average heart rate, day average heart rate and night average heart rate.
GW31-e0086
Yunyi Xie, Zheng Liu, Ling Zhang
Capital Medical University
OBJECTIVES Salt sensitivity of blood pressure (SSBP) increases the morbidity and mortality of cardiovascular disease and is an independent risk factor for cardiovascular events. SSBP is a complex health issue associated with genetic factors. At present, there is still a lack of research reports on the relationship between SSBP and genetic polymorphisms in non-coding region, especially micro-RNA (miRNA). This study aimed to identify the association between candidate single-nucleotide polymorphisms in miRNAs and SSBP in a Han Chinese population.
METHODS The study was based on a Cohort Study of Systems Epidemiology on Salt-Sensitivity of Blood Pressure (EpiSS) in two cities of north China. We conducted a population-based case-control study, 219 salt sensitivity and 543 salt resistant were recruited in the study. A modified Sullivan’s acute oral saline load and diuresis shrinkage test (MSAOSL-DST) was used to distinguish the person of salt-sensitive and salt-resistant. Medical history, lifestyle risk factors are obtained by questionnaire, while blood pressure and weight were measured by physical examination, and blood and urine specimens are collected. Candidate single nucleotide polymorphisms (SNPs) were summarized from two approaches: literature retrieval of association studies in candidate miRNAs related SNPs and miRNAs related SNPs in previous our study results. We used the Sequenom Mass ARRAY Platform to genotype the 24 single-nucleotide polymorphisms, and the genetic risk score (GRS) was used to evaluate the joint genetic effect.
RESULTS According to screening principles, we selected 30 coding miRNAs related SNPs (miRNAs-SNPs) for validation. Multiple logistic regression analysis showed that 6 miRNAs-SNPs (miR-1307-5p/rs11191676, miR-1307-5p/rs2292807, miR-145/rs41291957, miR-19a-3p/rs4284505, miR-382-5p/rs4906032 and miR-4638-3p/rs6601178) were associated with SSBP in general population. After adjusting for potential confounders, the six SNPs (miR-1307-5p/rs11191676, miR-1307-5p/rs2292807, miR-145/rs41291957, miR-4638-3p/rs6601178, miR-382-5p/rs4906032 and miR-15b-5/rs10936201) showed significant correlation. In acute salt loading process, after adjusting for the confounding factors of waist-to-hip ratio, hypertension, total cholesterol, baseline urinary sodium concentration and smoking, miR-15b-5p/rs10936201, miR-145/rs41291957, miR-4638-3p/rs6601178 were significantly associated with DBP elevation (P≤0.05); miR-145/rs41291957 and miR-382-5p/rs4906032 were significantly associated with the increase of MAP (P≤0.05); In the process of diuresis shrinkage, miR-15b-5p/rs10936201 was significantly correlated with the decrease of SBP (P≤0.05); rs11191676/miR-1307-5p, rs11191676/miR-1307-5p and miR-145/rs41291957 were significantly associated with DBP decrease (P≤0.05); rs10936201/miR-15b-5p, rs11191676/miR-1307-5p, rs2292807/miR-1307-5p, rs41291957/miR-145 were significantly associated with MAP decrease (P≤0.05). A weighted GRS was composed of 6 coding miRNAs-SNPs. In a multiple logistic regression analysis adjusted by sex, age and hypertension demonstrate that with rising 1 score for weighted GRS, the risk of SSBP increased to 2.663-fold (95% CI: 1.869–3.795).
CONCLUSIONS Seven SNPs in miRNAs are associated with salt sensitivity of blood pressure. GRS was performed to evaluate the joint genetic effect. With increasing of GRS, the risk of SSBP in different populations were increased. It played a certain role to systematically screening SNPs.
GW31-e0093
Peng Cai, Yan Peng, Xukai Wang
Daping Hospital, Army Medical University
OBJECTIVES This study aimed to explore the relationship of thyroid-stimulating hormone (TSH), free triiodothyronine (FT3), and free thyroxine (FT4) levels with hypertension subtypes.
METHODS One thousand fifty six euthyroid adults were included as research samples. They underwent measurement of clinic blood pressure and 24-h ambulatory blood pressure monitoring. Then, they were divided into normotension (NT), white coat hypertension (WCH), masked hypertension (MHT), and sustained hypertension (SHT) groups. The 24-h dynamic electrocardiogram was performed to analyze the heart rate variability (HRV), so as to reflect the cardiac autonomic function. The relationship between hypertension subtypes, thyroid function, and HRV was analyzed.
RESULTS The TSH concentration was significantly higher in the SHT group than in the NT group (P=0.001). The FT3 concentration was higher in the SHT group than in the NT and MHT groups (P=0.013, P=0.008), while the FT4 concentration was significantly higher in the WCH group than in the NT group (P=0.002). The changes in HRV were observed between the SHT, WCH, and MHT groups and the NT groups, as well as between the SHT and the MHT groups. The multiple linear regression analysis also showed that FT3, HRV (RMSSD and PNN50), and blood pressure levels linearly correlated with one another (P<0.05). Meanwhile, the linear regression analysis showed a linear negative correlation between FT4 and HRV (SDANN) in the WCH+NT group (P=0.001).
CONCLUSIONS Thyroid function was closely related to hypertension subtypes such as WCH probably due to the changes in the cardiac autonomic function.
GW31-e0094
Peng Cai, Xukai Wang
Department of Cardiology, Institute of Field Surgery, Daping Hospital, Army Medical University
OBJECTIVES This study aimed to investigate whether hypertension subtypes such as white-coat hypertension (WCHT), diagnosed with the addition of nighttime blood pressure (BP) criteria, were related to coronary artery stenosis (CAS) and cardiac arrhythmia.
METHODS In this cross-sectional observational study, 2106 adult participants were recruited in Daping Hospital between December 2017 and April 2019. After rigorous screening, 844 participants were selected who did not use antihypertensive, lipid-lowering and anti-platelet drugs. They were divided into normotensive (NT), WCHT, masked hypertension (MHT) and sustained hypertension (SHT) groups based on the results of clinic BP measurement and ambulatory BP monitoring. Coronary angiography and ambulatory electrocardiography were performed to determine the participants’ CAS and cardiac arrhythmia status.
RESULTS Coronary angiography revealed 555 patients with CAS and 288 patients with normal coronary arteries. Logistic regression analysis showed that the incidence of CAS was higher in the MHT and SHT groups than in the NT group, while no significant change was found in the WCHT group. The logarithm of Gensini score was used to compare the degree of CAS between the groups. Multiple linear regression analysis showed that the degree of CAS was higher in the WCHT, MHT and SHT groups than in the NT group. The incidence of frequent atrial premature beat, atrial tachycardia and ventricular cardiac arrhythmia were significantly higher in the WCHT and SHT groups than in the NT group, while only ventricular cardiac arrhythmia change was found in the MHT group. There was no difference in ventricular arrhythmias between the groups in participants with normal coronary arteries, while there was difference in atrial arrhythmias in the WCHT and SHT groups as compared to the NT group.
CONCLUSIONS This study found that hypertension subtypes such as WCHT were closely associated with CAS and cardiac arrhythmia.
GW31-e0172
Mekhman Mamedov1, Samir Mehdiyev2, Isakh Mustafayev2
1National Medical Research Center for Preventive Medicine, Moscow, Russia
2Azerbaijan State Advanced Training Institute for Doctors Named After A. Aliyev, Baku, Azerbaijan
OBJECTIVES To study the clinical features of hypertension in patients with DM2, depending on the level of glycemic control.
METHODS A cohort study included 528 patients aged 30–69 years (30.5% men, 69.5% women). Patients answered the questions of the ARIC questionnaire which include the duration, severity of hypertension, tactics and the form of antihypertensive therapy taken. The level of blood pressure (BP) was measured twice, in the sitting position with a 5-minute interval, and the average values were taken. Electrocardiographic (ECG) signs of left ventricular hypertrophy (LVH) were taken using the Sokolow-Lyon criteria and the Cornell voltage index, and echocardiographic (EchoCG) signs of LVH were determined in the presence of a left ventricular mass index in men >115 g/m2, and in women >95 g/m2. Glycohemoglobin values (HbA1c) ≥7% were regarded as inadequate control of glycemic status. Statistical analysis of the data was carried out using the discriminant method.
RESULTS The HbA1c level in the inadequate control stage was 50.3% higher than in the compensation stage (9.42 vs. 6.27%, respectively, P.
CONCLUSIONS Most of patients with poor glycemic control did not reach target BP. Treatment with blockers of the renin-angiotensin-aldosterone system was received by 24.0% of the respondents, and combination and long-term antihypertensive therapy was administered to only half of the subjects. It is necessary to strengthen the patients’ adherence to treatment, as well as measures aimed at improving the knowledge and skills of endocrinologists and cardiologists.
GW31-e0205
Haoyu Wang
Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College
OBJECTIVES Hyperuricemia and Hypertension are two independent risk factors of renal function damage. Our research aimed to investigate the synergistic interaction between hyperuricemia and hypertension toward reduced eGFR.
METHODS Our analyses included 11,694 participants from a cross-sectional population-based Northeast China Rural Cardiovascular Health Study. Interaction was assessed on both multiplicative and additive scales.
RESULTS The prevalence of reduced estimated glomerular infiltration rate (eGFR) was 2.11% in our population. After adjustment of age, sex, race, education level, family income, current smoking and drinking status, body mass index, total cholesterol, high-density lipoprotein cholesterol, and diabetes, subjects with both hyperuricemia and hypertension suffered from a 11.004 (95% CI: 7.080–17.102) times risk of reduced eGFR than the healthy reference group, greater than that in participants with only hyperuricemia (5.741, 95% CI: 3.045–10.825) or hypertension (1.145, 95% CI: 0.764–1.715). Furthermore, additive interaction between hyperuricemia and hypertension was statistically significant and synergistic (relative excess risk due to interaction: 5.118, 95% CI: 0.611–9.624; the attributable proportion due to interaction: 0.465, 95% CI: 0.151–0.779; Synergy index: 2.047, 95% CI: 1.017–4.120). However, our results revealed no significant interaction on the multiplicative scale.
CONCLUSIONS Hyperuricemia and hypertension may have a synergistic interaction toward renal function loss in addition to their independent impacts. Our findings may provide a straightforward illustration which is easy for the public to realize the hazard of coexistent hypertension and hyperuricemia on renal injury.
GW31-e0246
Huan Sun1, Xiaoping Li2, Yuhua Zhang2, Shengnan Liu2, Xiaochun Chen2
1Inner Mongolia Medical University
2Inner Mongolia Autonomous Region International Mongolian Medical Hospital
OBJECTIVES To investigate the effect of hyperhomocysteinemia on cognitive function in patients with hypertension
METHODS One hundred and eight primary hypertensive patients were selected from October 2018 to October 2019 in the outpatient clinic of Cardiovascular Medicine Department of International Mongolian Hospital of Inner Mongolia Autonomous Region. History of hypertension and related risk factors were collected and fasting plasma homocysteine, liver function, renal function, blood lipids and blood glucose were detected. The patients were divided into two groups according to the level of plasma Hcy. Those with Hcy ≥10 μmol/L were the experimental group (Hypertension combined with hyperhomocysteinemia group), and those with Hcy <10 μmol/L were the control group (Simple Hypertension Group). The cognitive function of patients was measured by mini-mental state examination (MMSE) and Montreal cognitive assessment (MoCA), and the differences in cognitive function and related influencing factors between the two groups were analyzed.
RESULTS There were no significant differences in hypertension grade, course of hypertension, smoking and drinking history between the two groups (P>0.05), and there were no significant differences in serum total cholesterol, low density lipoprotein, blood glucose, office systolic blood pressure and diastolic blood pressure between the two groups (P>0.05). The plasma Hcy level in the experimental group was significantly higher than that in the control group (17.02±13.13 μmol/L vs. 8.92±1.70 μmol/L, P<0.05), and the difference was statistically significant. The plasma triglycerides, creatinine and uric acid were higher than those in the control group (2.011±25 mmol/L vs. 1.66±0.91 mmol/L; 67.66±17.12 μmol/L vs. 60.22±13.08 μmol/L; 331.00±90.42 μmol/L vs. 281.14±59.28 μmol/L, all P<0.05); plasma high density lipoprotein was lower than control group (1.35±0.29 mmol/L vs. 1.53±0.42 mmol/L, P<0.05). The total score of MMSE in the experimental group was lower than that in the control group (25.94±3.88 vs. 27.57±1.81, P>0.05), and there was no statistical difference between the two groups. In the MMSE score, the language ability of the experimental group was lower than that of the control group (8.47±1.06 vs. 8.90±0.30, P<0.05), and the difference was statistically significant. The total score of cognitive function test group by MoCA was lower than that of control group (21.61±4.87 vs. 24.24±3.17, P<0.05), the difference was statistically significant, and the visuospatial and executive functions, attention and orientation were significantly lower than that of control group (2.94±1.37 vs. 4.05±1.01, 4.95±1.31 vs. 5.52±0.97, 5.64±0.69 vs. 5.90±0.30, all P<0.05). The correlation analysis showed that the total score of MoCA was negatively correlated with plasma Hcy level (r=-0.311 P=0.001), negatively correlated with age (r=-0.413 P=0.000), and positively correlated with years of education (r=0.617 P=0.000). The partial correlation controlled for years of education and age respectively, and plasma Hcy was negatively correlated with total score of MOCA (P<0.05). Multiple linear regression analysis of Hcy, years of education and age showed that Hcy was an independent risk factor for MoCA (B=-0.061 P=0.036).
CONCLUSIONS Hyperhomocysteinemia aggravates the decline of cognitive function in hypertensive patients; plasma Hcy is negatively correlated with total MoCA score and is an important risk factor affecting cognitive level; MoCA is more sensitive than MMSE in evaluating cognitive function.
GW31-e0380
Huan Wang1,2, Jingkang Zhu1,2, Huiwu Hong1,2, Canwang Wang1,2, Hui Chen1,2
1Hypertension Laboratory, Fujian Provincial Cardiovascular Disease, Fujian Provincial Hospital
2The Shengli Clinical Medical College of Fujian Medical University
OBJECTIVES CACNA1D encodes the pore-forming α1-subunit of Cav1.3, an L-type voltage-gated Ca2+-channel. The CACNA1D rs9810888 polymorphism was significantly associated with DBP and MAP. Therefore, the relationship between CACNA1D c.A920G gene mutant with hypertension was researched.
METHODS Human CACNA1D c.A920G gene mutant families were investigated. Immunofluorescence was used to detect the expression of CACNA1D in human endothelial cells (HUVECs) and human umbilical vein. HUVECs were transfected with liposome carrying CACNA1D wild-type gene and CACNA1D c.A920G, respectively. Cav1.3 and ET-1 protein expression were observed by Western blot. The contents of intracellular calcium were measured by Flou-4, and the effect of expression of ET-1 mRNA by RT-qPCR and Western blot. The CACNA1D c.A920G mutant rats were edited using genetic engineering.
RESULTS Human CACNA1D c.A920G germline mutant families were prone to hypertension. Cav1.3 l-type calcium channels were found in HUVECs and human umbilical vein. Compared with wildtype HUVECs, the intracellular Ca2+ (0.05364±0.00252 vs. 0.07105±0.00236, n=3 P<0.01), the protein of CAV1.3 (1.04±0.25 vs. 1.44±0.32, n=6, P<0.05) and ET-1 (1.32±0.54 vs. 2.43±0.98, n=8, P<0.01), and the mRNA expression of ET-1 (1.00±0.00 vs. 1.49±0.19, n=3, P<0.05) in HUVECs with CACNA1D c.A920G mutant were increased significantly. We first found that twelve human CACNA1D c.A920G mutant 28-week rats that had significant elevated blood pressure than ten wildtype 28-week rats (146.80±1.91 vs. 134.20±2.67, P<0.05).
CONCLUSIONS The CACNA1D c.A920G mutant was related with hypertension. This work was supported by the National Youth Science Foundation Project of China (81800363), the Provincial Natural Science Foundation of Fujian (2018J01240), Youth Innovation Project of Science and Technology Department of Fujian Province (20GJ05136), the Fujian Provincial Medical Innovation Project (2018CX4), the Funding Scheme for Young and Middle-aged Talents Development Projects (2018XQN24) in Fujian Provincial Health Family Planning Commission, and the Joint Fund for Research and Construction of High-Level Hospitals in Fujian Provincial Hospital (2017LHJJ03).
GW31-e0447
Gang Li, Yifang Guo, Qingjuan Zuo, Lili He, Yan Wang, Lu Zhang, Tingting Zhang, Qiuyan Wang, Yi Liang
Hebei General Hospital
OBJECTIVES Hypertension is closely related to atherosclerosis. Atherosclerosis is considered to be a chronic inflammatory condition. Carotid intima-media thickness (IMT) reflects the degree of apparent changes in atherosclerosis. Some studies have shown that sulfatide is closely related to inflammatory response and atherosclerotic lesions. In this study, we observed whether there was a correlation between serum sulfatide and carotid IMT in elderly patients with hypertension, and which index could more accurately reflect the progression of atherosclerotic lesions.
METHODS One hundred and fifteen elderly patients with hypertension admitted to our hospital from July 2018 to July 2019 were selected as the experimental group. According to blood pressure level, they were divided into mild hypertension group (n=35), moderate hypertension group (n=40) and severe hypertension group (n=40). In addition, 110 old people of the same age who underwent physical examination in the physical examination center of our hospital were selected as the control group. Firstly, the difference of serum sulfatide level and carotid IMT level between the experimental group and the control group was observed, and then the difference of serum sulfatide level and carotid IMT level between different hypertension groups was compared in the experimental group.
RESULTS Serum sulfatide level and carotid IMT in the experimental group were higher than that in the control group, and the difference was statistically significant (P<0.05). Moreover, with the increase of blood pressure, serum sulfatide level and carotid IMT in elderly patients with hypertension showed an increasing trend (r=0.389, P=0.021).
CONCLUSIONS Serum sulfatide level and carotid IMT in the experimental group were higher than that in the control group, and the difference was statistically significant (P<0.05). Moreover, with the increase of blood pressure, serum sulfatide level and carotid IMT in elderly patients with hypertension showed an increasing trend (r=0.389, P=0.021).
GW31-e0571
Zhipeng Zhang, Xiaoping Chen
West China Hospital, Sichuan University
OBJECTIVES As an emerging interventional technique to treat resistant hypertension, renal denervation (RDN) has also attracted considerable attention due to its potential beneficial effects on glucose and lipid metabolism. Given that inconsistent results documented among studies, we aimed to perform a systematic review and meta-analysis to further elaborate this issue.
METHODS The PubMed, EMBASE, Web of Science (SCI) and ClinicalTrials.gov databases were comprehensively searched from their inception date to June 18, 2020 for relevant clinical studies evaluating the efficacy of RDN on glucose and lipid levels. The outcomes of interest were changes in fasting glucose, insulin, C-peptide, hemoglobin A1C (HbA1C), homeostatic model assessment-insulin resistance (HOMA-IR), cholesterol and triglyceride (TG) levels before versus after RDN and also RDN versus control group. The mean differences (MD) of the outcomes measured before versus after RDN and RDN versus control group were pooled by a fixed or randomized effects model. Heterogeneity was quantified with chi-square (χ 2) and inconsistency index (I 2). Assessment of publication bias was performed by funnel plot and Egger’s test.
RESULTS A total of 1600 studies were initially identified and 20 of them (randomized controlled studies: 6; non-randomized controlled studies: 1; observational cohort studies: 13) involving 2277 subjects were included in the final analysis. No significant changes were observed after RDN in fasting glucose (weighted mean difference [WMD] -0.16 mmol/L; 95% CI -0.34, 0.02 mmol/L), insulin (standardized mean difference [SMD] 0.06; 95% CI -0.13, 0.26), C-peptide (SMD -0.06; 95% CI -0.25, 0.14), HbA1C (SMD -0.04; 95% CI -0.14, 0.06), HOMA-IR (SMD -0.12; 95% CI -0.35, 0.11), total cholesterol (TC) (WMD -0.11 mmol/L; 95% CI -0.37, 0.15 mmol/L), low-density lipoprotein cholesterol (LDL-C) levels (WMD -0.18 mmol/L; 95% CI -0.59, 0.24 mmol/L) during the follow-up. Changes of fasting glucose, insulin, HbA1C and TC levels in RDN groups were not significantly different from that in control group. High-density lipoprotein cholesterol (HDL-C) and TG were slightly but significantly improved after RDN (WMD 0.07 mmol/L, 95% CI 0.01, 0.14 mmol/L; WMD -0.26 mmol/L, 95% CI -0.51, -0.01 mmol/L, respectively). Funnel plot and Egger’s test demonstrated the absence of potential publication bias.
CONCLUSIONS Catheter-based RDN appeared to have no impact on glucose metabolism. There was a statistically significant but clinically negligible improvement in HDL-C and TG levels on the basis of the current whole body of evidences. Future researches with more rigorous designs are warranted to draw definitive conclusions.
GW31-e0594
Jindong Wan1,2, Peijian Wang1,2
1Department of Cardiology, The First Affiliated Hospital, Chengdu Medical College, Chengdu, Sichuan 610500, PR China
2Key Laboratory of Aging and Vascular Homeostasis of Sichuan Higher Education Institutes, Chengdu, Sichuan 610500, PR China
OBJECTIVES To evaluate the safety and feasibility of bilateral adrenal venous sampling (AVS) via femoral vein using single 5F Tig catheter with preliminary experience.
METHODS One hundred and six consecutive patients diagnosed with primary aldosteronism underwent AVS via femoral vein using single 5F Tig catheter. 5F Tig catheters were properly used catheter molding technology during AVS. The result of success rate of bilateral AVS, operate time, and operative complications were analyzed. Based on operation time versus sequence of cases, the learning curve was plotted.
RESULTS All patients were sequentially cannulated into bilateral adrenal veins with a single 5F Tig catheter. The success rate of bilateral AVS was 90.6% (96/106). The mean operation time was (33±8) min, mean fluoroscopy time was (5.8±1.7) min, average X-ray dose was (117.3±25.5) mGy, and dosage of contrast media was (17.3±5.5) mL. One patient (0.9%) experienced a hematoma at the femoral vein puncture, while other patients were not occurred any complications. The learning curve showed that the operation time shortened as the number of cases increased, and about 33 cases could cross the learning curve.
CONCLUSIONS AVS via femoral vein using single shaped 5F Tig catheter is safe and technically feasible, with a high success rate of bilateral AVS. The required number of operating procedures to the technique was approximate 33, and single 5F Tig catheter can be a preferred method of saving consumables when bilateral AVS via femoral vein.
GW31-e0635
Shuyuan Zhang, Jun Cai, Weili Zhang
State Key Laboratory of Cardiovascular Disease, Hypertension Center, FuWai Hospital, National Center for Cardiovascular Diseases, Peking Union Medical College & Chinese Academy of Medical Sciences, Beilishi Road 167, Xicheng District, Beijing 100037, China
OBJECTIVES A novel coronavirus disease 2019 (COVID-19) is currently breaking out around the world and greatly threatening the people’s health. Along with the appearance of psychological stress such as anxiety, depression and other negative emotions caused by this special circumstance, hypertensive patients may experience large fluctuations in blood pressure, especially for the elderly. However, it is uncertain that whether the home BP monitored by smartphone-based application (app) can be controlled well during the epidemic.
METHODS The home BP data were longitudinally collected via the app from October 21st, 2019 to March 21st, 2020 (before and during the COVID-19 outbreak in China) of 7394 hypertensive patients aged 60–80 years who were enrolled from 42 hospitals at 23 provinces/municipalities in the strategy of blood pressure intervention in the elderly hypertensive patients (STEP) study (ClinicalTrials.gov, NCT03015311). The fluctuation of home morning BP was compared in Wuhan, the epidemic center, and in other areas of China. The mental disorder-anxiety was assessed by the Generalized Anxiety Disorder Scale.
RESULTS The variation of average morning systolic BP (SBP) of patients in Wuhan was significantly higher than in other areas of China by 1.8–3.9 mmHg (all P<0.05). In Wuhan, the morning SBP, compared with 132 (9.4) mmHg before the epidemic (Oct 21st–Nov 20th, 2019), increased by an average of 3 mmHg during the outbreak, while reduced to 131.9 (10.3) mmHg again during the epidemic control period (Feb 21st–Mar 21st, 2020). In other areas of China, the morning SBP of patients fluctuated less than 1 mmHg, with an average of 131 mmHg. The logs of doctors or patients accessing the BP module in the app were analyzed, and the results showed that the frequency of app visits by doctors during the epidemic period was consistent with the fluctuation of patient’s morning SBP. During Nov 21st, 2019–Feb 20th, 2020, since doctors were engaged in control of the COVID-19 epidemics, the frequency of checking the patients’ BP by doctors dramatically fell to 34% per month and then nearly to zero in the most serious days in Wuhan, and consequentially, during the epidemic control period (Feb 21st–Mar 21st, 2020), the numbers of BP visits grew back to 16% per month. In addition, the frequency of checking BP by patients through the app continuously increased when compared with the non-epidemic period, while it was in particular higher in Wuhan than in other areas of China. Compared with those without anxiety, the morning SBP of patients with mild anxiety increased by about 6–7 mmHg in Wuhan and 2 mmHg on average in other areas of China, respectively (P<0.05).
CONCLUSIONS Our findings showed that the epidemic of COVID-19 could cause a short-term remarkably increase of morning SBP among elderly hypertensive patients in Wuhan. Using smartphone-based apps for managing home BP is feasible for BP control during the special epidemic period.
GW31-e0669
Shiqun Chen, Zhiping Gao, Jin Liu, Zhidong Huang, Yong Liu
Guangdong Provincial People’s Hospital
OBJECTIVES The risk assessment of blood pressure failure in community hypertension patients can help identify high-risk patients and carry out active blood pressure Control measures, the current lack of uncontrolled hypertension rate study and its predictive factors of hypertension patients receiving anti-hypertensive treatment in community clinics. Therefore, the study exploring the risk predictors of controlled hypertension among patients in the community receiving anti-hypertensive therapy and build a risk prediction model for it.
METHODS We were consecutively selected in February 2018 and March 2018 in Guangdong Province, and received 1089 patients ≥18 years old, and finally included 1039 patients in the analysis and collected the socio-demographic information of hypertension patients, disease awareness, hypertension management, and the use and demand of mobile health tools in patients with hypertension in the community. The controlled blood pressure of patients is the main end point, which is defined as patients self-reporting most families/offices. Blood pressure is measured below 140/90 mmHg. This study compared the socio-demographic information, disease awareness, and the use of mobile health tools among hypertensive patients in the community between the blood pressure compliance and non-compliance groups. The variables with P<0.05 in Logistic regression multivariate analysis are independent risk factors that affect the blood pressure of hypertension patients. The risk factors that are important in univariate analysis can be used to select the final prediction model for blood pressure failure. The modeling data set of 906 community hypertension patients was used, and the internal verification was performed by bootstrap method for 100 times.
RESULTS The average age was 61±13 years old, there were 549 (53.9%) males, the average body mass index was 24.3±3.2 kg/m2. Multivariable analysis showed that: full-time work (Ratio, OR: 1.98, 95% confidence interval, CI: 1.46-–2.69), self-financed medical care (OR: 3.47, 95% CI: 2.08–5.80), non-marital status (OR: 2.01, 95% CI: 1.35–3.27), poor knowledge of hypertension diagnosis (OR: 3.28, 95% CI: 2.42–4.45), poor drug compliance (OR: 1.51, 95% CI: 1.08–2.11), poor knowledge of hypertension treatment (OR: 2.94, 95% CI: 2.16–3.99) and reluctance to remind blood pressure measurement information (OR: 1.64, 95% CI: 1.08–2.50) were identified independent predictors of uncontrolled hypertension among patients in the community who received anti-hypertensive treatment, the risk model includes the following seven factors: full-time work (4 points), self-pay medical care (7 points), non-marital status (4 points), poor cognitive diagnosis of hypertension (6 points), poor drug compliance (3 points), poor cognition of hypertension treatment (6 points) and unwilling reminder of blood pressure measurement (3 points), Hosmer Lemeshow statistics of multivariate models do not suggest lack of appropriateness (χ2=6.5649, P=0.5842). In the verification data set, the risk score showed good discriminating ability and predicting ability for the incidence of uncontrolled hypertension (C-static: 0.771) by bootstrap method.
CONCLUSIONS We have established 7 key predictors based prediction model with good predictability and high discriminatory ability. Our prediction model provide an good evaluation tool for community hypertension prevention and treatment to identify high-risk populations of uncontrolled hypertension.
GW31-e0671
Shiqun Chen, Xiaoling Cen, Zhidong Huang, Yong Liu
Guangdong Provincial People’s Hospital
OBJECTIVES Investigate the status of nonadherence in patients with hypertension and their requirements for the function of mobile health tools.
METHODS A total of 1089 patients with hypertension in the community were investigated by questionnaire. Nonadherence was defined as the monthly proportion of days covered <90%.
RESULTS Patients with poor adherence accounted for 26.4%. Significantly related factors were less than 65 years old (OR: 0.549, 95% CI: 0.313–0.963), single (OR: 0.599, 95% CI: 0.361–0.993), inaccurate recognition of the diagnostic value of hypertension (OR: 0.657, 95% CI: 0.439–0.983), failure to purchase antihypertensive drugs at community health stations (OR: 0.642, 95% CI: 0.424–0.972), poor blood pressure control (OR: 0.391, 95% CI: 0.254–0.602), irregularity of blood pressure measuring (OR: 0.523, 95% CI: 0.351–0.780) and smoking (OR: 1.586, 95% CI: 1.018–2.469). The patients had a large demand for functions such as doctor-patient communication, hypertension knowledge, and changes in blood pressure over a week.
CONCLUSIONS Enhancing the patient’s medication habits may improve the patient’s drug nonadherence. Mobile health tools can help patients strengthen their medication habits, but the exploration of their functions needs more verification.
GW31-e0701
Yilin Chen, Tingyan Xu, Jiguang Wang
The Shanghai Institute of Hypertension, Department of Hypertension, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China
OBJECTIVES We investigated right ventricular (RV) function using speckle tracking echocardiography (STE) in patients with primary aldosteronism (PA).
METHODS Our study included 51 PA patients and 50 age- and gender-matched primary hypertensive patients. We performed 2-dimensional echocardiography to measure cardiac structure and function. We performed STE offline analysis on RV four-chamber (RV4CLS) and free wall longitudinal strains (RVFWLS).
RESULTS PA patients, compared with primary hypertensive patients, had a significantly (P=0.045) larger left ventricular (LV) mass index (112.0±22.6 g/m2 vs. 95.8±18.5 g/m2) and left atrial volume index (26.9±6.0 mL/m2 vs. 24.7±5.6 mL/m2) and higher prevalence of LV concentric hypertrophy (35.3 vs. 12.0%), although they had similarly normal LV ejection fraction (55–77%). PA patients also had a significantly (P=0.047) larger right atrium and ventricle, lower tricuspid annular plane systolic excursion, and higher E/E′t (the peak early filling velocity of trans-tricuspid flow to the peak early filling velocity of lateral tricuspid annulus ratio), estimated pulmonary arterial systolic pressure and RV index of myocardial performance. On RV strain analysis, PA patients had a significantly (P<0.001) lower RV4CLS (-18.1±2.5% vs. -23.3±3.4%) and RVFWLS (-21.7±3.7% vs. -27.9±4.5%) than primary hypertensive patients. Overall, RV4CLS and RVFWLS were significantly (r=-0.58 to -0.41, P<0.001) correlated with plasma aldosterone concentration and 24-h urinary aldosterone excretion. After adjustment for confounding factors, the associations for RV4CLS and RVFWLS with 24-h urinary aldosterone excretion remained significant, with a standardized coefficient of -0.48 and -0.55, respectively (P<0.001).
CONCLUSIONS In addition to LV abnormalities, PA patients also show impaired RV function, probably because of adrenal aldosterone hypersecretion.
GW31-e0702
Wei Zhang, Jiguang Wang
Department of Cardiovascular Medicine, Centre for Epidemiological Studies and Clinical Trials, Shanghai Key Laboratory of Hypertension, The Shanghai Institute of Hypertension, Department of Hypertension, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China
OBJECTIVES We investigated ambulatory blood pressure (BP) in relation to hyperuricemia, dietary sodium intake and their interaction in children and adolescents with hypertension.
METHODS The 616 study participants were 10–24 years old and had primary hypertension diagnosed after admission in a specialized inpatient ward. Ambulatory BP monitoring was performed during hospitalization. 24-hour urine was collected for measurements of electrolytes. Hyperuricemia was defined as a serum uric acid of ≥327.25 μmol/L in patients <18 years old and of ≥420 μmol/L and ≥360 μmol/L, respectively, in male and female patients ≥18 years old.
RESULTS In adjusted analyses, patients with hyperuricemia (n=283), compared with those with normal serum uric acid, had similar 24-h systolic BP (131.7 mmHg, P=0.54) and a significantly (P=0.005) lower 24-h diastolic BP (77.5 vs. 80.9 mmHg) and higher 24-h pulse pressure (54.2 vs. 51.7 mmHg). In similar adjusted analyses, 24-h ambulatory pulse pressure, but not systolic/diastolic BP (P=0.12), significantly differed across the quartile distributions of urinary sodium excretion (P for trend ≤0.04). Further adjusted analyses showed significant (P=0.04) interaction between serum uric acid and urinary sodium excretion in relation to 24-h systolic BP. In patients with hyperuricemia (P=0.04), but not those with normal serum uric acid (P=0.13), 24-h systolic BP was significantly associated with urinary sodium excretion, with a 6.5±2.1 mmHg difference between quartiles 4 and 1. Similar results were observed for daytime and nighttime BP and pulse pressure.
CONCLUSIONS Both hyperuricemia and higher dietary sodium intake were associated with higher pulse pressure, and their interaction further heightened systolic BP.
GW31-e0703
Wei Zhang, Jiguang Wang
Centre for Epidemiological Studies and Clinical Trials, Shanghai Key Laboratory of Hypertension, The Shanghai Institute of Hypertension, Department of Hypertension, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China
OBJECTIVES We investigated associations of blood pressure (BP) with albuminuria and left ventricular hypertrophy (LVH) in young, middle and older aged patients with hypertension and/or diabetes mellitus.
METHODS Study participants were treated patients with hypertension or diabetes, enrolled in a China nationwide registry. The 2510 patients were classified into young (<45 years, n=345), middle (45–64 years, n=1383) and older (≥65 years, n=782) age groups. Clinic BP was measured three times consecutively on each of the two clinic visits. These six readings were averaged for analyses. Albuminuria was defined as a urinary albumin-to-creatinine ratio of ≥30 mg/g. LVH was assessed by the electrocardiogram (ECG) Cornell product and voltage methods.
RESULTS The prevalence of albuminuria and ECG-LVH was 17.8 and 6.5%, respectively. Mean (±SD) systolic/diastolic BP was 132.0±16.5/85.2±11.9 mmHg, 136.8±17.9/81.7±11.2 mmHg, and 139.8±16.7/75.8±10.4 mmHg in the young, middle and older age groups. In the young age group, the prevalence of albuminuria increased from 8.8% in systolic/diastolic BP <120/80 mmHg to 14.6, 16.0 and 16.5% in 120–129/80–84, 130–139/85–89 and ≥140/90 mmHg, respectively. The corresponding values were 8.9, 7.0, 18.1 and 22.2%, respectively, in the middle age group, and 21.2, 15.5, 16.4 and 24.4%, respectively, in the older age group. Analyses adjusted confirmed the J-shaped relation between BP and albuminuria in the older but not young age group. The prevalence of ECG-LVH was significantly (P for trend ≤0.04) higher with increasing BP similarly in all age groups.
CONCLUSIONS The relationship between BP and organ damage seems to be age-dependent for albuminuria but not ECG-LVH.
GW31-e0724
Di Zhang, Yan Li, Qifang Huang, Jiguang Wang
Centre for Epidemiological Studies and Clinical Trials, The Shanghai Institute of Hypertension, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China
OBJECTIVES In a 12-week, randomized, controlled trial, we investigated whether home blood pressure monitoring (HBPM) would improve treatment adherence and blood pressure control in stage 2–3 hypertension.
METHODS Eligible patients (18–75 years old and a systolic/diastolic blood pressure of 160–199/100–119 mmHg after 1-week wash-out) were randomized in a 1:4 ratio to HBPM and a control group without HBPM. All patients started antihypertensive treatment with the irbesartan 150 mg/hydrochlorothiazide 12.5 mg/day combination, with the possible addition of irbesartan 150 mg/day and up-titration to irbesartan 300 mg/hydrochlorothiazide 25 mg/day at 4 and 8 weeks of follow-up, respectively. The primary endpoint was the clinic blood pressure control (systolic/diastolic, non-diabetes <140/90 mmHg and diabetes <130/80 mmHg) rate at 12 weeks of follow-up.
RESULTS The randomized patients in the HBPM (n=96) and control groups (n=405) had similar characteristics at baseline, and similar use of higher dosages of irbesartan/hydrochlorothiazide (300 mg/12.5–25 mg) at 4 (9.4 vs. 12.2%, P=0.45) and 8 weeks of follow-up (27.1 vs. 35.5%, P=0.13). During follow-up, both the treatment discontinuation rate (1.0 vs. 12.6%, P=0.0008) and less optimal compliance rate (<90% of prescribed medication, 1.0 vs. 9.9%, P=0.005) were significantly lower in the HBPM than control group. The proportion of patients who achieved the goal of clinic blood pressure control at 12 weeks of follow-up was significantly higher in the HBPM than control group (66.7 vs. 55.1%, P=0.04).
CONCLUSIONS In conclusion, HBPM improved treatment adherence and blood pressure control in patients with hypertension, in spite of similar antihypertensive treatment intensity.
GW31-e0911
Yueyuan Liao, Jianjun Mu
First Affiliated Hospital of Medical College, Xi’an Jiaotong University, Xi’an 710000, China
OBJECTIVES Arterial stiffening is an important independent risk factor for cardiovascular disease and events. Obesity and hypertension are closely related to arterial stiffness and play an important role in the development of arterial stiffness. The impact of the long-term burden of BMI and BP from childhood to adulthood on arterial stiffness in adults, especially between sexes, still need to be explored. We aimed to examine the impact of the cumulative long-term burden and trends of BMI and BP from childhood on adult arterial stiffness and explore sex differences in this association.
METHODS This study is based on the Hanzhong Adolescent Hypertension Cohort, an ongoing prospective study. This longitudinal study consisted of 1553 individuals aged 6–15 years who were examined 4 or more times for BMI and BP since childhood, with a follow-up period of 30 years. Growth curves of BMI and BP, measured repeatedly at multiple time points from childhood, were constructed using a random-effects model. The area under the curve (AUC) was calculated as a measure of the long-term burden (total AUC) and trends (incremental AUC) of BMI and BP from childhood. Brachial–ankle pulse wave velocity (baPWV) was recorded by a non-invasive automatic waveform analyser in adulthood. Statistical significance was set as a two-tailed P value of less than 0.05.
RESULTS An analysis of the subjects’ general characteristics found that males had higher height, weight, BMI, SBP, DBP, waist circumference, hip circumference, total cholesterol, low-density lipoprotein cholesterol, triglycerides, fasting glucose, serum uric acid, urine albumin and baPWV than females. The results from the simple correlation analysis showed that baPWV in adults was significantly correlated with BMI total AUC (r=0.12), BMI incremental AUC (r=0.05), SBP total AUC (r=0.45), SBP incremental AUC (r=0.26), DBP total AUC (r=0.38), DBP incremental AUC (r=0.19). The total effect of the total AUC and incremental AUC of BMI and BP on adult baPWV were analyzed by linear regression models. Adult baPWV was associated with SBP total AUC (standardized regression coefficient, β=6.45, P<0.001), SBP incremental AUC (β=2.60, P<0.001), DBP total AUC (β=7.60, P<0.001) and DBP incremental AUC (β=1.57, P<0.001). And there were no gender differences in the effect of total AUC and incremental AUC of BP on adult baPWV. There was no association between the total AUC of BMI and arterial stiffness regardless of sex. However, there were sex differences in the association between the incremental AUC of BMI and arterial stiffness (P=0.019 for interaction). The incremental AUC of BMI indicated an increased risk of arterial stiffening during adulthood in males, but this association was not found in females.
CONCLUSIONS In summary, we demonstrated that the cumulative long-term burden and incremental trends of BP are all significantly associated with adult arterial stiffness measured as baPWV. In addition, there were differences in the impact of long-term trends of BMI from childhood to adulthood on the development of arterial stiffness between males and females. The long-term growth trends of BMI can increase the risk of arterial stiffness in males, independent of BP trends, but not in females. These results emphasize the importance of developing prevention and intervention strategies for hypertension and obese men (especially those who are gradually gaining weight) to reduce the risk of arterial stiffening and cardiovascular disease in adulthood.
GW31-e0996
Yumeng Shi, Lihua Hu, Minghui Li, Congcong Ding, Wei Zhou, Tao Wang, Lingjuan Zhu, Huihui Bao, Xiaoshu Cheng
The Second Affiliated Hospital of Nanchang University
OBJECTIVES We aimed to evaluate the relation of the ankle-brachial index (ABI) with the risk of stroke and to examine any possible effect modifiers among hypertensive patients without atrial fibrillation.
METHODS A total of 10,750 subjects with hypertension aged 27–96 years was included in the current study. The outcome was a stroke. Odds ratios of stroke concerning ABI were calculated using multivariate logistic regression models.
RESULTS Among 10,750 hypertensive participants, 690 (6.42%) had a stroke. Multivariate logistic analyses showed that ABI was negatively correlated with ABI was negatively correlated with the risk of stroke (Per SD increment; adjusted OR, 0.88; 95% CI, 0.82–0.94). Compared with participants in Q 1, the odds ratios (95% CI) for those in the Q2 (1.05–1.10), Q3 (1.10–1.15) and Q4 (≥1.15) were 0.71 (0.56, 0.90), 0.87 (0.70, 1.08) and 0.81 (0.65, 1.01), respectively. However, compared with higher ABI value, lower ABI value (<1.05) would significantly increase the risk of stroke [OR: 1.26, 95% CI (1.05–1.50)], especially in the elderly over 65 years old. A generalized additive model and a smooth curve fitting showed that there existed an L-shaped association between ABI and the risk of stroke.
CONCLUSIONS Our results suggest that an L-shaped association between ABI and risk of stroke was found in general hypertensive patients, with a turning point at about 1.05. Compared with higher ABI value, lower ABI value (<1.05) would significantly increase the risk of stroke [OR: 1.26, 95% CI (1.05–1.50)], especially in the elderly over 65 years old.
GW31-e1180
Qiong Ma, Yu Yan, Jianjun Mu
The First Affiliated Hospital of Xi’an Jiaotong University
OBJECTIVES Hypertension is the most common preventable risk factor for cardiovascular disease and is associated with an increased risk of all-cause morbidity. Elevated blood pressure (BP) affects approximately 270 million people in China and represents a public health crisis nationwide. Although considerable progress has been made on effective antihypertensive medications, current control rates for hypertension remain low in China, especially in rural areas. Controlling blood pressure and therefore reducing the risk of cardiovascular events remains a serious challenge. In this study, we aim to provide an evidence-based approach to the management of hypertension in rural China.
METHODS A parallel cluster randomized controlled trial was undertaken in 24 rural villages in Hanzhong, Shaanxi, China. Eligible villages were randomized using a computer-generated randomization method. After baseline evaluation, we enrolled 2116 hypertensive participants over 40 years of age who met inclusion criteria in this study. Participants of intervention villages (9 clusters) were offered intensive anti-hypertensive treatment, including the use of antihypertensive medication, healthy lifestyle guidance, and intensive BP monitoring by the village doctors. All major classes of antihypertensive agents were provided at low cost to the participants. BP measurements data were recorded by a mobile device and uploaded to the monitoring center, and medications were adjusted on a monthly basis to target a BP of less than 130/80 mmHg. Participants in the control villages (11 clusters) were advised to continue with their regular treatment. Participants were seen monthly for the first 3 months and every 6 months thereafter. Hypertension control was defined as SBP <140 mmHg and DBP <90 mmHg, and an intensive control rate (SBP <130 and DBP <80 mmHg) of hypertension were also estimated according to the 2017 American College of Cardiology/American Heart Association High Blood Pressure Guideline.
RESULTS At baseline, the mean age was 65.6±9.5 years and there was no significant difference in mean BP between the intervention and control groups (SBP, 153.7±16.3 vs. 154.9±16.1 mmHg; DBP, 86.7±11.1 vs. 85.2±10.7 mmHg; all P>0.05). After 18 months, there was a significant higher rate of the antihypertensive treatment in the intervention group compared with the control group (94.0 vs. 73.62%, P<0.05). Importantly, the mean SBP was 126.5±11.6 mmHg and DBP was 71.0±9.1 mmHg in the intervention group, which was significantly lower than that in the control group (145.3±19.0 and 78.6±11.2 mmHg; all P<0.05). The intervention group also presented a higher control rate of hypertension, both at 140/90 mmHg (86.1%) and 130/80 mmHg (68.6%) standard, compared with the control group (38.3 and 17.4%, respectively; all P<0.05).
CONCLUSIONS The internet-based integrated management approach can significantly reduce blood pressure and improve the control rate of hypertension and is expected to provide an effective approach for BP controlling in rural China.
GW31-e1201
Zhihua Zhang, Qin Luo, Nanfang Li
Hypertension Center of People’s Hospital Of Xinjiang Uygur Autonomous Region, Xinjiang Hypertension Institute, National Health Committee Key Laboratory of Hypertension Clinical Research
OBJECTIVES Metabolic syndrome (MetS) is a complex disorder comprising obesity, hyperglycemia, dyslipidemia, and hypertension, which is associated with a greater risk of cardiovascular disease. In a recent meta-analysis of 31 studies, comprising 3838 patients with primary aldosteronism (PA) and 9284 patients with essential hypertension, PA had increased risk of (OR 1.53, 95% CI 1.22–1.91) than patients with essential hypertension. PA has two main subtypes: unilateral, largely represented by aldosterone producing adenoma (APA), and bilateral, usually idiopathic hyperaldosteronism (IHA). We aimed to compare the prevalence of MetS in patients with the two subtypes of PA, diagnosed using adrenal venous sampling (AVS), and to determine the factors associated with the presence of MetS in PA patients.
METHODS This was a retrospective cross-section study in a single center. We included one hundred and sixty-nine hypertensive patients who had been diagnosed with PA in hypertension center of People’s Hospital of Xinjiang Autonomous Region between January and December 2017. We analyzed metabolic parameters from 169 PA patients subtyped by AVS, including 85 unilateral PA patients and 84 bilateral PA patients, and we also included 169 non-PA patients matched for age and sex.
RESULTS The prevalence of MetS in patients with PA was higher than in patients without PA, but this difference was not statistically significant ((72.2 vs. 65.7%, P>0.05). Patients with unilateral PA had higher concentrations of aldosterone and lower serum potassium than patients with bilateral PA. However, patients with bilateral PA had higher prevalence of MetS (79.8 vs. 64.7%, P=0.029), obesity (40.5 vs. 24.7%, P=0.029), dyslipidemia (72.6 vs. 55.3%, P=0.019) and hyperglycemia (29.8 vs. 16.5%, P=0.040) than those with unilateral PA. Meanwhile, compared to unilateral PA, patients with bilateral PA had higher BMI (27.6±4.6 vs. 25.6±3.3 kg/m2 P=0.001), waist circumference (98.5±11.4 vs. 93.3±10.6 cm, P=0.003) and fasting plasma glucose (4.98±1.16 vs. 4.64±0.93 mmol/L, P=0.034). We conducted logistic regression analysis to clarify the association between bilateral PA and MetS/obesity. Bilateral PA was significantly associated with MetS (OR, 2.150; 95% CI, 1.074–4.302; P=0.031) and obesity (OR, 2.072; 95% CI, 1.073–4.001; P=0.030). The associations remained statistically significant after adjustment for age, sex and duration of hypertension.
CONCLUSIONS Patients with bilateral PA have a higher prevalence of MetS than those with unilateral PA, despite unilateral PA patients exhibiting higher concentrations of aldosterone and lower serum potassium, suggesting that unilateral PA and bilateral PA may have differing mechanisms of MetS.
GW31-e1207
Tilakezi Tuersun, Qin Luo, Nanfang Li
Hypertension Center of People’s Hospital Of Xinjiang Uygur Autonomous Region, Xinjiang Hypertension Institute, National Health Committee Key Laboratory of Hypertension Clinical Research, Urumqi 830001, Xinjiang, China
OBJECTIVES Patients with primary aldosteronism (PA) have increased risk of target-organ damage, among which vascular calcification is an important indicator of cardiovascular mortality. A recent study indicated that the abdominal aorta calcification (AAC) prevalence was almost 1-fold higher in patients with PA than controls with essential hypertension, matched for age, sex, and blood pressure. Unilateral and bilateral PA are the most common subtypes of PA. However, no studies have addressed the difference in the prevalence of AAC between the two subtypes. In addition to aldosterone, parathyroid hormone (PTH), an important regulator of calcium metabolism, was also reported to be elevated in individuals with unilateral PA. Therefore, we hypothesized that the prevalence of AAC may be higher in individuals with unilateral PA, which may be related to the plasma aldosterone concentration (PAC) and PTH levels.
METHODS This was a single-center, cross-sectional study conducted in the Hypertension Center of People’s Hospital of Xinjiang Uygur Autonomous Region. From January 2017 to January 2018. A total of 156 patients diagnosed with PA who underwent AVS were included in our study, among whom, 76 were diagnosed with unilateral PA and 80 were diagnosed with bilateral PA. We also included 156 with essential hypertension (EH) matched by age and sex. The aortic calcification index (ACI) presented the severity of AAC and was measured by adrenal computed tomography scan and measured in all subjects.
RESULTS Patients with PA had higher PAC, PTH, and 24 h urinary calcium levels than EH patients, whereas serum potassium and TC levels were lower than those in the EH group (P<0.05). The prevalence of AAC was significantly higher in patients with PA than those with EH (32.7 vs. 19.6%; P=0.013). Moreover, the degree of ACI was more severe in PA patients than in EH patients (4.32±3.61% vs. 2.53±2.42%, P=0.028). In the PA subgroup analysis, unilateral PA was associated with a higher prevalence and more severe AAC than bilateral PA (40.7 vs. 25.0%; 5.12±4.07% vs. 3.08±2.34%, respectively). Moreover, PAC and PTH levels were higher in individuals with unilateral PA than in those with bilateral PA (P<0.05). After adjusted for age, age, duration of hypertension, abdominal circumference, phosphorus, etc., multivariate regression analysis revealed that PAC and PTH were positively-associated with AAC in patients with PA (OR 1.235, 95% CI 1.100–1.373, P<0.001; OR 1.038, 95% CI 1.013–1.064, P=0.002, respectively).
CONCLUSIONS Unilateral PA patients exhibited a higher prevalence of AAC and more severe AAC due to elevated PAC and PTH levels.
GW31-e1222
Jianshu Chen, Jing Yu
Lanzhou University Second College of Clinical Medicine
OBJECTIVES Reversing left ventricular hypertrophy (LVH) can reduce the incidence of adverse cardiovascular events. However, there is no clear superiority-inferiority differentiation between angiotensin-converting enzyme inhibitors (ACEI), angiotensin receptor blockers (ARB), angiotensin receptor neprilysin inhibitors (ARNI), beta-blockers (BB), calcium channel blockers (CCB), and diuretics in reversing LVH in hypertensive patients.
METHODS To provide further evidence for choosing the optimal antihypertensive drug for improving LVH, we performed a network meta-analysis of randomized controlled trials (RCTs) based on the Cochrane library database, Embase, and PubMed, and identified 49 studies involving 5402 patients that were eligible for inclusion.
RESULTS It was found that ARB could improve LVH in hypertensive patients more effectively than CCB (MD −4.05, 95% CI −7.81 to −0.34) and BB (MD −4.52, 95% CI −7.80 to −1.13). Matched comparison of renin-angiotensin system inhibitors (RASi) showed that the effect of ACEI in reducing left ventricular mass index (LVMi) was not effective as that of ARB (MD −3.68, 95% CI −7.30 to −0.15). The surface under the cumulative ranking for each intervention indicated that the use of ARB was more effective among the six types of antihypertensive drugs (98%). This network meta-analysis revealed that the use of ARB in antihypertensive therapy could achieve better efficacy in reversing LVH in hypertensive patients.
CONCLUSIONS This network meta-analysis revealed that the use of ARB in antihypertensive therapy could achieve better efficacy in reversing LVH in hypertensive patients.
GW31-e1269
Ting Wu1,2,3, Qing Zhu1,2,3, Bin Zhu1,2,3, Shasha Liu1,2,3, Shanshan Liu1,2,3, Nanfang Li1,2,3
1Hypertension Center of People’s Hospital of Xinjiang Uygur Autonomous Region
2Xinjiang Hypertension Institute
3National Health Committee Key Laboratory of Hypertension Clinical Research
OBJECTIVES Hypertension induced with vasculitis leads to more serious damage to the target organs. Pentraxin-3 (PTX-3) derived from the secretion of macrophages, neutrophils, endothelial cells, epithelial cells and vascular smooth muscle cells, which expressed locally at the sites of inflammatory processes and regulate the immune activity of macrophages. The objectives of our study were to investigate the serum PTX-3 levels in vasculitis (Vas) and analyze this correlation with hypertension.
METHODS A total of 155 cases consisting 51 patients with Vas [including 7 cases of takayasu arteritis (TA), 24 cases of polyarteritis nodosa (PAN), and 20 cases of antineutrophil cytoplasmic antibody-associated Vas (AAV)] were screened by angiography and/or biopsy; 46 patients with essential hypertensions (EH) and 58 healthy controls (HC) were enrolled in this study. Serum PTX-3 levels were determined by enzyme-linked immunosorbent assay.
RESULTS Compared with the HC and EH, the serum PTX-3 levels in systemic Vas were significantly higher (both P<0.001, 4.42±0.95 vs. 2.67±0.92, 4.42±0.95 vs. 2.95±0.60), and there was no significant difference between HC and EH (P=0.886, 2.67±0.92 vs. 2.95±0.60). There was no significant difference of PTX-3 levels among TA, PAN, and AAV, as well as active and inactive groups, and renal and nonrenal groups, respectively. There was no significant correlation between PTX-3 levels and blood pressure, erythrocyte sedimentation rate, or Birmingham Vasculitis Activity Score. Receiver operating characteristic analysis has shown that the best cutoff point was at 3.618 ng/mL; the sensitivity and specificity were calculated as 84.3 and 93.5% for the diagnosis of Vas from heath control, and the best cutoff point was at 3.425 ng/mL, The sensitivity and specificity were calculated as 88.2 and 82.6% for the diagnosis of Vas from essential hypertension.
CONCLUSIONS Serum PTX-3 levels were significantly higher in patients with Vas than essential hypertension or health control, and elevated PTX-3 levels can help identify Vas patients from healthy or essential hypertensive populations.
GW31-e1280
Xiaotong Wang, Nanfang Li
Hypertension Center of People’s Hospital of Xinjiang Uygur Autonomous Region; Xinjiang Hypertension Institute, National Health Committee Key Laboratory of Hypertension Clinical Research, Urumqi 830001 Xinjiang, China
OBJECTIVES Our study aimed to analyze the effect of SPL compliance on endothelial dysfunction by assessing MAU in patients with PA.
METHODS The study included 145 confirmed PA patients who received long-term medical treatment (mean, 5 years). We assigned patients who took SPL continuously as the compliant group (N=102) and those who withdrew from SPL treatment for at least 6 months as the noncompliant group (N=43). The primary outcome was the prevalence of MAU and secondary were clinical outcomes include serum potassium, blood pressure and the defined daily doses of antihypertensive agents.
RESULTS Expectedly, compliance with SPL treatment better improved patients’ blood pressure and serum potassium levels. Patients with PA who complied fully with SPL treatment had a lower rate of MAU than did the noncompliant patients (13.7 vs. 34.9%, respectively; P=0.004). Multivariate logistic regression analyses adjusted for age and sex showed that continuous SPL treatment was associated with a lower presence of MAU (odds ratio,0.319; 95% confidence interval, 0.135–0.750; P=0.009). This association remained significant after further adjusting for other major risk factors. However, in the subgroup analysis, the protective effect against MAU was limited in compliant patients treated with ≥40 mg/day SPL compared with that of the noncompliant patients (9.6 vs. 34.9%, P<0.05).
CONCLUSIONS Our findings demonstrated that in addition to improving high blood pressure and hypokalemia, full compliance with the appropriate dose of SPL may benefit endothelial function as reflected by a lower prevalence of MAU in patients with PA.
GW31-e1306
Shasha Liu, Qing Zhu, Ting Wu, Guoliang Wang, Xintian Cai, Ayiguzaili Aihemaiti, Xiayire Aierken, Nanfang Li
Hypertension Center of People’s Hospital of Xinjiang Uygur Autonomous Region
OBJECTIVES Systemic vasculitis is a group of unexplained connective tissue diseases with non infectious inflammation and necrotizing vasculitis as the basic pathological changes. Its clinical manifestations are nonspecific and often lead to multiple system invasion and multiple organ failure. It can also lead to hypertension or malignant hypertension. But it is difficult to diagnose. In recent years, related studies have found that MCP-1 is related to inflammation and immune response. Monocyte chemotactic protein 1 (MCP-1), which is able to adjust the migration of monocytes/macrophages, osmosis and raise monocytes and T lymphocyte cells to participate in a variety of inflammation, are closely linked to various types of systematic vasculitis. However, serum MCP-1 levels have not been evaluated in patients with systemic vasculitis, who also have moderate hypertension. The study was performed to investigate the level of serum MCP-1 in determining the patients.
METHODS We reviewed all patients admitted to our center for the etiology screening of hypertension between January 2013 and December 2016. All vasculitis are diagnosed by a rheumatologist and fulfilled the American College of Rheumatology (ACR) 1990 criteria. Serum samples were collected in 43 patients with systemic vasculitis with hypertension, 46 patients with essential hypertension (EH) and 43 healthy controls (HC). Serums MCP-1 were measured using commercially available ELISA kits.
RESULTS The serum MCP-1 levels were significantly higher in patients with systemic vasculitis, compared with EH (134.65 (73.74, 262.75) pg/mL, 68.43 (65.42, 104.84) pg/mL, P<0.008) and HC (134.65 (73.74, 262.75) pg/mL, 59.1 (37.41, 90.18) pg/mL, P=0.001), and no difference regarding serum MCP-1 levels could be found between EH and HC (P=0.197). Furthermore, it was significantly increasing in patients with renal than non-renal involvement (196.16 (104.41, 310.35) pg/mL, 73.74 (41.24, 145.95) pg/mL, P=0.001) and HC (P<0.001). However, there was no significant statistical differences between active and inactive phase. Serum MCP-1 levels with patients in systemic vasculitis were positive correlation with serum creatinine levels (r=0.387, P<0.010), 24-hour proteinuria (r=0.3627, P<0.0297) and. white blood cell counts (r=0.365, P<0.016).
CONCLUSIONS Serums MCP-1 were significantly higher in patients with systemic vasculitis compared with EH and HC, especially in patients with renal involvement. Serum MCP-1 might be as a potential biomarker tools in the systemic vasculitis with renal involvement.
GW31-e1359
Fengyu Han, Feng Hu, Xiaoshu Cheng
Department of Cardiovascular Medicine, the Second Affiliated Hospital of Nanchang University, Nanchang of Jiangxi, China
OBJECTIVES Serum uric acid (SUA) is a product of human purine metabolism. Most studies revealed that high level of SUA was associated with stroke risk and indicated that the influence of SUA on stroke is due to the secondary association of SUA with other established etiological risk factors, including hypertension, arterial stiffness, obesity, hyperinsulinemia. We propose that hypertension may be an intermediate in the pathway between hyperuricemia and stroke. However, the previous results concerning the role of SUA as an independent risk factor for stroke in hypertensive patients were still controversial. Our aim was to investigate the association between SUA level and the risk of first stroke in Chinese adults with hypertension.
METHODS This prospective study enrolled 14, 268 participants and was conducted from July 2018 to July 2023 in Wuyuan, Jiangxi province of China. After excluding 34 individuals without hypertension at baseline, 7 cases lost to follow-up, 7 cases without serum uric acid data, 191 cases with atrial fibrillation, 1371 cases with eGFR ≤60 mL/min/1.73 m2, and 817 cases with prior stroke at baseline, finally 11, 841 Chinese adult hypertensive individuals (median follow-up time: 614 days) were included in our analysis. Computed tomography or magnetic resonance imaging as well as medical records were conducted to confirm first stroke events (ischemic stroke, hemorrhagic stroke or unspecified stroke). Multivariate linear regression models and Kaplan-Meier curves were used to examine the associations between SUA level or HUA (hyperuricemia) and first stroke events.
RESULTS The clinical baseline characteristics of this cohort (age: 62.95±9.14 years, range 27–93 years; male, 45.67%) were presented by the quartiles of baseline SUA level, and the prevalence of HUA was 52.23%. A total of 99 (0.84%) first stroke events (51 ischemic events, 15 hemorrhagic events and 33 unspecified stroke events) occurred. There were no statistically significant differences in various stroke events between groups by the quartiles of baseline SUA level (P>0.05). The risk of first stroke was not significantly associated with the increased SUA levels or HUA. This unrelated association was also found after adjustment for gender and age. Subgroup analyses found that this association between SUA level and total first stroke events was modified by aging (age ≥60 years): in group with age less than 60 years, subjects with HUA had an statistically significant higher risk of total first stroke events compared to population without HUA (adjusted-HR: 4.89, 95% CI 1.36–17.63, P=0.015). However, this positive association was disappeared in aging population (adjusted-HR: 0.97, 95% CI 0.60–1.56, P=0.886; P-value for interaction=0.043). Survival analysis further confirmed this discrepancy (Kaplan–Meier, log-rank P=0.013 for non-aging group, P=0.899 for aging group, respectively). There were not statistically significant interactions in any of the subgroups, including gender, mean arterial pressure tertiles, systolic blood pressure tertiles, diastolic blood pressure tertiles, antihypertensive drugs usage, body mass index tertiles and central obesity.
CONCLUSIONS No significant evidence in present study indicates that increased SUA levels or HUA are predictive of first stroke in Chinese adults with hypertension. Nonetheless, subgroup analyses found that this correlation was modified by aging.
GW31-e1380
Haoyu Wang
Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College
OBJECTIVES AHA’s Life’s Simple 7 cardiovascular health score is recommended for use in primary prevention. Simpler tools not requiring laboratory tests, such as the Fuster-BEWAT score (FBS) (blood pressure [B], exercise [E], weight [W], alimentation [A], and tobacco [T]), are also available. This study sought to compare the effectiveness of Life’s Simple 7 and FBS in predicting the newly proposed 4-tiered LVH classification based on LV dilatation (high LV end-diastolic volume [EDV] index) and concentricity (mass/end-diastolic volume [M/EDV]0.67) in the general Chinese population.
METHODS Participants from Northeast China Rural Cardiovascular Health study who underwent cardiac echocardiography (n=11,261) were enrolled. Patients with LVH were divided into 4 groups – eccentric nondilated (normal M/EDV and EDV), eccentric dilated (increased EDV, normal M/EDV), concentric nondilated (increased M/EDV, normal EDV), and concentric dilated (increased M/EDV and EDV) – and compared with patients with normal LVM.
RESULTS With poor Life’s Simple 7 and FBS as references, individuals with ideal Life’s Simple 7 and FBS showed lower adjusted odds of having eccentric nondilated (Life’s Simple 7, odds ratio [OR]: 0.26; 95% confidence interval [CI]: 0.20–0.34 vs. FBS, OR: 0.28; 95% CI: 0.20–0.38), eccentric dilated (OR: 0.73 [0.57–0.94] vs. OR: 0.57 [0.43–0.76]), concentric nondilated (OR: 0.12 [0.04–0.38] vs. OR: 0.19 [0.07–0.52]), and concentric dilated LVH (OR: 0.12 [0.03–0.37] vs. OR: 0.26 [0.10–0.72]). Similar levels of significantly discriminating accuracy were found for Life’s Simple 7 and FBS with respect to the eccentric nondilated (C-statistic: 0.737; 95% CI: 0.725–0.750 vs. 0.731; 95% CI: 0.718–0.744, respectively), eccentric dilated (0.684 [0.670–0.699] vs. 0.686 [0.671–0.701]), concentric nondilated (0.658 [0.624–0.692] vs. 0.650 [0.615–0.684]), and concentric dilated LVH (0.711 [0.678–0.744] vs. 0.698 [0.663–0.733]).
CONCLUSIONS Our findings demonstrate that the FBS appears capable of performing just as well as does the Life’s Simple 7 in predicting the novel 4-group classification of LVH, making the FBS particularly suited as a reliable low-cost indicator of CV health in settings where access to laboratory analysis is limited and health care resources are constrained.
ARRHYTHMIAS
GW31-e0068
Jue Wang, Suyun Liu
Second Hospital of Hebei Medical University
OBJECTIVES Although several electrocardiography (ECG) criteria have been purposed to distinguish left and right origins of outflow tract ventricular arrhythmias (OT-VAs) with lead V3 transition, they remain limited in clinical practice. The purpose of this study was to determine if lead I R-wave amplitude is effective to distinguish left and right origin.
METHODS We measured lead I R-wave amplitude in a retrospective cohort of 82 OT-VAs patients with lead V3 transition and positive complex in lead I who underwent successful catheter ablation from right ventricular outflow tract (RVOT) and left ventricular outflow tract (LVOT). The optimal R-wave threshold was identified and diagnostic indices were compared with V2S/V3R, transitional zone (TZ) index, and the V2 transition ratio.
RESULTS Lead I R-wave amplitude for LVOT origins was significantly higher than that of RVOT origins (0.55±0.13 vs. 0.32±0.15 mV; P<0.001). The area under the curve (AUC) for the lead I R-wave amplitude by receiver operating characteristic (ROC) analysis was 0.926, with a cut-off value of ≥0.45 predicting an LVOT origin with a 92.9% sensitivity and 88.2% specificity. In accuracy, lead I R-wave amplitude was superior to V2S/V3R, TZ index, and the V2 transition ratio. 92.3% LVOT cases exhibiting lead I R-wave amplitude ≥0.45 mV originated from the right coronary cusp (RCC) and the left and right coronary cusp junction (L-RCC).
CONCLUSIONS The R-wave amplitude in lead I provides a useful and simple criterion to identify RCC or L-RCC origin in OT-VAs with lead V3 transition.
GW31-e0070
Yuhong Peng, Yanzhuo Ma, Dongmei Wang
The 980st Hospital of the PLA Joint Logistics Support Force (Bethune International Peace Hospital of PLA)
OBJECTIVES To observe the feasibility of applying a new full-lead wearable ambulatory ECG based on fabric electrodes for non-invasive long-range batch ECG monitoring in physical training.
METHODS Fifty-four subjects were selected, wearing a new ambulatory ECG monitoring suit for 48 hours and performing physical training, observing their tolerance and equipment reliability, and comparing the information (heart rate, arrhythmia and ST segment changes) obtained by ambulatory ECG monitoring before and after the training.
RESULTS Forty-four subjects completed the whole process of ambulatory ECG monitoring. The average record time is 43.44±3.56 h. All subjects indicated that the suit is comfortable or bearable. The real-time monitoring signal has clear graphics and reliable quality. The incidence of arrhythmia was compared before and after the training. The incidence of atrial arrhythmia on the day of the training was significantly lower than before the training (P=0.001). The average heart rate, total heart rate, and maximum heart rate on the day of the training are higher than before. The standard deviation of NN intervals (SDNN), SD of 5 min N–N intervals (SDANN), and SDNN index are also higher than the normal working state (P<0.05). The slowest heart rate, root mean square of successive RR interval differences (RMSSD), percentage of successive RR intervals that differ by more than 50 ms (PNN50), and frequency domain indicators did not differ between groups (P>0.05).
CONCLUSIONS The new wearable full-lead ambulatory electrocardiogram monitoring devices can be used for long-term batch ECG monitoring in physical training. Appropriate intensity training can reduce the occurrence of arrhythmia and improve heart rate variability. e, total heart rate, and maximum heart rate on the day of the training are higher than before. The standard deviation of NN intervals (SDNN), SD of 5 min N-N intervals (SDANN), and SDNN index are also higher than the normal working state (P<0.05). The slowest heart rate, root mean square of successive RR interval differences (RMSSD), percentage of successive RR intervals that differ by more than 50 ms (PNN50), and frequency domain indicators did not differ between groups (P>0.05).
GW31-e0071
Yuhong Peng, Yanzhuo Ma, Dongmei Wang
The 980st Hospital of the PLA Joint Logistics Support Force (Bethune International Peace Hospital of PLA)
OBJECTIVES To investigate the effects of long-term high intensity training on cardiovascular in military personnel.
METHODS High-intensity training soldiers were divided into long-term high-intensity group (127 cases) and short-term control group (87 cases) according to the training time. The data of blood pressure and electrocardiogram and the incidence of arrhythmia were compared in 5 km weight-bearing training. Phosphocreatine kinase isoenzyme (CKMB), troponin T (TnT), adrenocorticotropic hormone (ACTH), adrenaline (ADR), norepinephrine (NADR) and angiotensin II (ANG2) were collected from 45 individuals.
RESULTS The diastolic blood pressure measured immediately after exercise in both groups is lower than before training (P<0.01). The P-wave width in II, RV5 amplitude (mv), and RV5+SV1 (mv) voltage of the long-term high-intensity training group were higher than those of the short-term control group no matter before or after training (P<0.01). There was no statistically significant difference in the incidence of atrial premature beats, short PR interval, first-degree AVB, complete right bundle branching and ST-T changes (P>0.05). The elevated levels of TnT, ACTH, ADR and ANG2 after training in the long-term high-intensity training group were lower than that in the short-term high-intensity training group (P<0.01).
CONCLUSIONS High-intensity military training leads to the potential risk to the heart of soldiers. The body’s heart structure undergoes adaptive changes after long-term training, and the stress response can be adjusted adaptively.
GW31-e0072
Na Wang, Lianjun Gao, Yunlong Xia, Xiaomeng Yin, Xianjie Xiao, Zhengyan Wang, Rongfeng Zhang
The First Affiliated Hospital of Dalian Medical University
OBJECTIVES By comparing the different initial heparin doses in patients with atrial fibrillation (AF) ablation who continuously took new oral anticoagulants (NOACs) and warfarin, the appropriate initial heparin dose in radiofrequency ablation for patients with NOACs was obtained.
METHODS This study was a single-center, randomized, double-blind, prospective clinical trial, which was selected from 2018.06 to 2019.05 in 187 patients with AF who underwent the first catheter radiofrequency ablation at the First Affiliated Hospital of Dalian Medical University and met the inclusion criteria. Patients were divided into warfarin (WG) group (38 cases) and new oral anticoagulants (NG110, NG120, NG130) group (149 cases) according to different anticoagulation schemes before surgery. The warfarin group was given an initial heparin dose of 100 U/kg as a control group, while the NOACs group was randomly divided into three groups using an SPSS random number generator and, respectively given an initial heparin dose of 110, 120, and 130 U/kg. Monitor the ACT every 15 minutes during the operation and achieve the target ACT value of 250–350 s. To observe the baseline ACT, average ACT compliance rate, and ACT at each time period in the Chinese Farin group and the new oral anticoagulant group, and the incidence of bleeding and thromboembolic complications during ablation, post