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      Polysaccharide extracts of Astragalus membranaceus and Atractylodes macrocephala promote intestinal epithelial cell migration by activating the polyamine-mediated K + channel

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          Abstract

          Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 µmol·L −1, SPD), alpha-difluoromethylornithine (2.5 mmol·L −1, DFMO), 4-Aminopyridine (40 µmol·L −1, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L −1), DFMO plus SPD, or DFMO plus polysaccha-ride extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca 2+ ([Ca 2+] cyt) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca 2+] cyt and accelerated migration of IEC-6 cells, compared with the controls ( P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca 2+] cyt, but also restored IEC-6 cell migration to control level ( P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased ( P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.

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          Physiological roles and properties of potassium channels in arterial smooth muscle.

          This review examines the properties and roles of the four types of K+ channels that have been identified in the cell membrane of arterial smooth muscle cells. 1) Voltage-dependent K+ (KV) channels increase their activity with membrane depolarization and are important regulators of smooth muscle membrane potential in response to depolarizing stimuli. 2) Ca(2+)-activated K+ (KCa) channels respond to changes in intracellular Ca2+ to regulate membrane potential and play an important role in the control of myogenic tone in small arteries. 3) Inward rectifier K+ (KIR) channels regulate membrane potential in smooth muscle cells from several types of resistance arteries and may be responsible for external K(+)-induced dilations. 4) ATP-sensitive K+ (KATP) channels respond to changes in cellular metabolism and are targets of a variety of vasodilating stimuli. The main conclusions of this review are: 1) regulation of arterial smooth muscle membrane potential through activation or inhibition of K+ channel activity provides an important mechanism to dilate or constrict arteries; 2) KV, KCa, KIR, and KATP channels serve unique functions in the regulation of arterial smooth muscle membrane potential; and 3) K+ channels integrate a variety of vasoactive signals to dilate or constrict arteries through regulation of the membrane potential in arterial smooth muscle.
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            Botanical polysaccharides: macrophage immunomodulation and therapeutic potential.

            Botanical polysaccharides exhibit a number of beneficial therapeutic properties, and it is thought that the mechanisms involved in these effects are due to the modulation of innate immunity and, more specifically, macrophage function. In this review, we summarize our current state of understanding of the macrophage modulatory effects of botanical polysaccharides isolated from a wide array of different species of flora, including higher plants, mushrooms, lichens and algae. Overall, the primary effect of botanical polysaccharides is to enhance and/or activate macrophage immune responses, leading to immunomodulation, anti-tumor activity, wound-healing and other therapeutic effects. Furthermore, botanical and microbial polysaccharides bind to common surface receptors and induce similar immunomodulatory responses in macrophages, suggesting that evolutionarily conserved polysaccharide structural features are shared between these organisms. Thus, the evaluation of botanical polysaccharides provides a unique opportunity for the discovery of novel therapeutic agents and adjuvants that exhibit beneficial immunomodulatory properties.
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                Author and article information

                Journal
                CJNM
                Chinese Journal of Natural Medicines
                Elsevier
                1875-5364
                20 September 2018
                : 16
                : 9
                : 674-682
                Affiliations
                1Pi-wei Institute, Guangzhou University of Chinese Medicine, Guangzhou 510405, China
                2College of Traditional Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510006, China
                Author notes
                *Corresponding author: LI Ru-Liu, E-mail: lrl@ 123456gzucm.edu.cn

                ΔThese authors contributed equally to this work.

                These authors have no conflict of interest to declare.

                Article
                S1875-5364(18)30107-9
                10.1016/S1875-5364(18)30107-9
                Copyright © 2018 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.
                Funding
                Funded by: National Natural Science Foundation of China
                Award ID: 30772753
                Award ID: 81173254
                Award ID: 81673940
                Funded by: Science and Technology Program of Guangzhou
                Award ID: 20160701335
                This work was supported by the National Natural Science Foundation of China (Nos. 30772753, 81173254, and 81673940), the Science and Technology Program of Guangzhou (No. 20160701335) and the First-class discipline construction major project of Guangzhou University of Chinese Medicine (Guangzhou University of Chinese Medicine Planning, 2018-No. 6).

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