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      Ribose-cysteine protects against the development of atherosclerosis in apoE-deficient mice

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          Abstract

          Ribose-cysteine is a synthetic compound designed to increase glutathione (GSH) synthesis. Low levels of GSH and the GSH-dependent enzyme, glutathione peroxidase (GPx), is associated with cardiovascular disease (CVD) in both mice and humans. Here we investigate the effect of ribose-cysteine on GSH, GPx, oxidised lipids and atherosclerosis development in apolipoprotein E-deficient (apoE-/-) mice. Female 12-week old apoE-/- mice (n = 15) were treated with 4–5 mg/day ribose-cysteine in drinking water for 8 weeks or left untreated. Blood and livers were assessed for GSH, GPx activity and 8-isoprostanes. Plasma alanine transferase (ALT) and lipid levels were measured. Aortae were quantified for atherosclerotic lesion area in the aortic sinus and brachiocephalic arch and 8-isoprostanes measured. Ribose-cysteine treatment significantly reduced ALT levels ( p<0.0005) in the apoE-/- mice. Treatment promoted a significant increase in GSH concentrations in the liver ( p<0.05) and significantly increased GPx activity in the liver and erythrocytes of apoE-/-mice ( p<0.005). The level of 8-isoprostanes were significantly reduced in the livers and arteries of apoE-/- mice ( p<0.05 and p<0.0005, respectively). Ribose-cysteine treatment showed a significant decrease in total and low density lipoprotein (LDL) cholesterol ( p<0.05) with no effect on other plasma lipids with the LDL reduction likely through upregulation of scavenger receptor-B1 (SR-B1). Ribose-cysteine treatment significantly reduced atherosclerotic lesion area by >50% in both the aortic sinus and brachiocephalic branch ( p<0.05). Ribose-cysteine promotes a significant GSH-based antioxidant effect in multiple tissues as well as an LDL-lowering response. These effects are accompanied by a marked reduction in atherosclerosis suggesting that ribose-cysteine might increase protection against CVD.

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          Most cited references30

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          N-acetylcysteine: multiple clinical applications.

          N-acetylcysteine is the acetylated variant of the amino acid L-cysteine and is widely used as the specific antidote for acetaminophen overdose. Other applications for N-acetylcysteine supplementation supported by scientific evidence include prevention of chronic obstructive pulmonary disease exacerbation, prevention of contrast-induced kidney damage during imaging procedures, attenuation of illness from the influenza virus when started before infection, treatment of pulmonary fibrosis, and treatment of infertility in patients with clomiphene-resistant polycystic ovary syndrome. Preliminary studies suggest that N-acetylcysteine may also have a role as a cancer chemopreventive, an adjunct in the eradication of Helicobacter pylori, and prophylaxis of gentamicin-induced hearing loss in patients on renal dialysis.
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            Vitamin E suppresses isoprostane generation in vivo and reduces atherosclerosis in ApoE-deficient mice.

            Oxidative modification of low density lipoprotein (LDL) has been implicated in atherogenesis. Evidence consistent with this hypothesis includes the presence of oxidized lipids in atherosclerotic lesions, the newly discovered biological properties conferred on LDL by oxidation and the acceleration of atherogenesis by in vivo delivery of the gene for 15-lipoxygenase, an oxidizing enzyme present in atherosclerotic lesions. However, it is still unknown whether oxidative stress actually coincides with the evolution of the disease or whether it is of functional relevance to atherogenesis in vivo. Isoprostanes are products of arachidonic acid catalyzed by free radicals, which reflect oxidative stress and lipid peroxidation in vivo. Elevation of tissue and urinary isoprostanes is characteristic of human atherosclerosis. Here, deficiency in apolipoprotein E in the mouse (apoE-/-) resulted in atherogenesis and an increase in iPF2alpha-VI, an F2-isoprostane, in urine, plasma and vascular tissue. Supplementation with vitamin E significantly reduced isoprostane generation, but had no effect on plasma cholesterol levels in apoE-/- mice. Aortic lesion areas and iPF2alpha-VI levels in the arterial wall were also reduced significantly by vitamin E. Our results indicate that oxidative stress is increased in the apoE-/- mouse, is of functional importance in the evolution of atherosclerosis and can be suppressed by oral administration of vitamin E.
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              A population at risk. Prevalence of high cholesterol levels in hypertensive patients in the Framingham Study.

              In the Framingham Study, coronary heart disease developed in every fifth man and every 17th woman by the age of sixty. The level of total cholesterol proved to be an excellent predictor of coronary heart disease in those aged less than 50 years. However, in those aged over 50 years, more accurate predictors of coronary heart disease risk were serum lipoprotein measurements, including low-density lipoproteins, very-low-density lipoproteins, very-low-density lipoprotein triglycerides, and high-density lipoproteins. Both low-density and very-low density lipoproteins have a linear association with coronary heart disease. On multivariate analysis, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol are independently related to coronary heart disease risk in both sexes. In women, but not in men, very-low-density lipoprotein cholesterol or the triglyceride level is an independent risk factor on multivariate analysis. By likelihood ratio analysis, high-density lipoprotein is shown to be the most powerful single factor for predicting coronary heart disease risk in both sexes relative to the lipid fractions. It appears that one of the most reliable profiles in this regard is the ratio of total cholesterol to high-density lipoprotein cholesterol. However, a special constellation of elevated triglycerides, low high-density lipoprotein levels, and "normal" cholesterol values should no longer be overlooked in assessing coronary heart disease risk. Both systolic and diastolic blood pressures are also related to risk of coronary heart disease in a linear fashion: the higher the level of pressure, the greater the incidence of coronary heart disease. Blood pressure and serum cholesterol are correlated with an r factor of 0.12, suggesting that those with higher blood pressure values tend to have higher serum cholesterol levels. Most physicians agree that treatment is advisable in those with cholesterol levels above 300 mg/dl; some believe therapy is necessary in those with levels of more than 250 mg/dl. Few realize that half of the patients in whom coronary heart disease will eventually develop have cholesterol values under 250 mg/dl. The National Institutes of Health Consensus Development Conference on Lipid Lowering has recommended that cholesterol levels be reduced to 200 mg/dl in all persons. Practicing physicians argue that coronary heart disease does not develop in most patients with cholesterol levels between 200 and 250 mg/dl. The problem lies in deciding which patients with these cholesterol levels actually have a lipid abnormality.(ABSTRACT TRUNCATED AT 400 WORDS)
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Project administrationRole: SoftwareRole: Writing – original draftRole: Writing – review & editing
                Role: Investigation
                Role: Formal analysisRole: InvestigationRole: Software
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: SoftwareRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: Software
                Role: Formal analysisRole: InvestigationRole: Methodology
                Role: Formal analysisRole: InvestigationRole: Methodology
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SoftwareRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                21 February 2020
                2020
                : 15
                : 2
                : e0228415
                Affiliations
                [1 ] Department of Biochemistry, School of Biomedical Sciences, University of Otago, Dunedin, New Zealand
                [2 ] Department of Surgical Sciences, Dunedin School of Medicine, University of Otago, Dunedin, New Zealand
                [3 ] Centre for Free Radical Research, Department of Pathology and Biomedical Science, University of Otago Christchurch, Christchurch, New Zealand
                [4 ] Metabolomics Australia, Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Australia
                Max Delbruck Centrum fur Molekulare Medizin Berlin Buch, GERMANY
                Author notes

                Competing Interests: The funder Max International provided support in the form of a salary for one of the author's conducting the research [TK] and some of the research materials including ribose-cysteine for which they own the patent and commercial license. The funder did not have any role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. The commercial affiliation with Max International does not alter our adherence to PLOS ONE policies on sharing data and materials. The authors do not have any other competing interests in form of consultancy, patents, products in development, or marketed products, etc.

                [¤]

                Current address: Cancer Genomics Program, The Sir Peter MacCallum Cancer Centre, Melbourne, Australia

                Author information
                http://orcid.org/0000-0002-1463-7593
                http://orcid.org/0000-0001-9857-0703
                Article
                PONE-D-19-25760
                10.1371/journal.pone.0228415
                7034848
                32084149
                c48e6167-c644-4a60-8a11-4c0b1958d59d
                © 2020 Kader et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 13 September 2019
                : 14 January 2020
                Page count
                Figures: 4, Tables: 1, Pages: 15
                Funding
                This study was supported by funding from Max International, LLC, Salt Lake City, UT, USA who provided the ribose-cysteine and by an Otago School of Medical Sciences Bequest Fund. ST was supported by an Otago Postgraduate Research Scholarship.
                Categories
                Research Article
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
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                Animal Studies
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                Medicine and Health Sciences
                Vascular Medicine
                Atherosclerosis
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                Custom metadata
                The underlying results required to replicate the study can be found in the paper and additional files containing supplementary and original data.

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