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Hydrogen decreases athero-susceptibility in apolipoprotein B-containing lipoproteins and aorta of apolipoprotein E knockout mice

  • Author Footnotes
    1 Equal contribution to this study.
    Guohua Song
    Footnotes
    1 Equal contribution to this study.
    Affiliations
    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Author Footnotes
    1 Equal contribution to this study.
    Hua Tian
    Footnotes
    1 Equal contribution to this study.
    Affiliations
    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Shucun Qin
    Correspondence
    Corresponding author at: Institute of Atherosclerosis, TaiShan Medical University, No. 2 YingSheng East Road, ShanDong 271000, PR China. Tel.: +86 538 6237252; fax: +86 538 6225275.
    Affiliations
    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Xuejun Sun
    Correspondence
    Corresponding author at: Department of Diving Medicine, Second Military Medical University, 800 Xiangyin Road, Shanghai 200433, PR China. Tel.: +86 21 63520020; fax: +86 21 63520020.
    Affiliations
    Department of Diving Medicine, Second Military Medical University, Shanghai, China
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  • Shutong Yao
    Affiliations
    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Chuanlong Zong
    Affiliations
    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Yingying Luo
    Affiliations
    Yangzhou University, JiangSu, China

    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Jia Liu
    Affiliations
    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Yang Yu
    Affiliations
    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Hui Sang
    Affiliations
    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Xinnong Wang
    Affiliations
    Key Laboratory of Atherosclerosis in Universities of Shandong and Institute of Atherosclerosis, TaiShan Medical University, ShanDong, China
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  • Author Footnotes
    1 Equal contribution to this study.

      Abstract

      Objective

      It is to characterize the underlying molecular mechanisms of the anti-atherosclerotic effects of hydrogen (dihydrogen; H2), a novel antioxidant. In particular, to examine the effects of hydrogen on athero-susceptibility in lipoproteins and aorta of apolipoprotein E knockout (apoE−/−) mice.

      Methods and results

      Plasma analysis by enzymatic method and spectrophotometric measurement showed that eight weeks intraperitoneally injection of hydrogen-saturated saline remarkably decreased plasma total and non-high-density lipoprotein (non-HDL) cholesterol, and malondialdehyde in apoE−/− mice fed either chow or high fat diet. Western blot analysis showed hydrogen treatment reduced the contents of apolipoprotein B (apoB), a major protein constituent of non-HDL in either plasma or hepatic tissues. Moreover, ELISA assay revealed that the production of tumor necrosis factor-α and interleukin-6 were significantly suppressed by hydrogen in RAW264.7 macrophages, after stimulation with the isolated non-HDL from treated or untreated mice. Immunohistochemistry of aortic valve sections revealed that hydrogen suppressed the expression of several proinflammatory factors and decreased vessel wall infiltration of macrophages. Besides, real-time PCR and Western blot analysis disclosed that hepatic scavenger receptor class B type I (SR-BI), ATP-binding cassette (ABC) transporters ABCG8, ABCB4, ABCB11, and macrophage SR-BI, were all induced by hydrogen treatment. Finally arterial wall lipid disposition displayed by oil red O staining was reduced significantly in aortic root and whole aorta en face in hydrogen administrated mice. In addition, hydrogen significantly improved HDL functionality in C57BL/6J mice assessed in two independent ways, namely (i) stimulation of cholesterol efflux from macrophage foam cells by measuring HDL-induced [3H]cholesterol efflux, and (ii) protection against LDL oxidation as a measure of Cu2+-induced TBARS formation.

      Conclusion

      These results reveal that administration of hydrogen-saturated saline decreases athero-susceptibility in apoB-containing lipoprotein and aortic atherosclerosis in apoE−/− mice and improves HDL functionality in C57BL/6J mice.

      Keywords

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      References

        • Libby P.
        Inflammation in atherosclerosis.
        Nature. 2002; 420: 868-874
        • Barter P.J.
        • Caulfield M.
        • Eriksson M.
        • et al.
        Effects of torcetrapib in patients at high risk for coronary events.
        N Engl J Med. 2007; 357: 2109-2122
        • Sniderman A.
        • McQueen M.
        • Contois J.
        • et al.
        Why is non-high-density lipoprotein cholesterol a better marker of the risk of vascular disease than low-density lipoprotein cholesterol?.
        J Clin Lipidol. 2010; 4: 152-155
        • Wang C.Y.
        • Chang T.C.
        Non-HDL cholesterol level is reliable to be an early predictor for vascular inflammation in type 2 diabetes mellitus.
        J Clin Endocrinol Metab. 2004; 89: 4762-4767
        • Ohsawa I.
        • Ishikawa M.
        • Takahashi K.
        • et al.
        Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals.
        Nat Med. 2007; 13: 688-694
        • Kajiyama S.
        • Hasegawa G.
        • Asano M.
        • et al.
        Supplementation of hydrogen-rich water improves lipid and glucose metabolism in patients with type 2 diabetes or impaired glucose tolerance.
        Nutr Res. 2008; 28: 137-143
        • Gharib B.
        • Hanna S.
        • Abdallahi O.M.
        • et al.
        Anti-inflammatory properties of molecular hydrogen: investigation on parasite-induced liver inflammation.
        C R Acad Sci III. 2001; 324: 719-724
        • Ohsawa I.
        • Nishimaki K.
        • Yamagata K.
        • et al.
        Consumption of hydrogen water prevents atherosclerosis in apolipoprotein E knockout mice.
        Biochem Biophys Res Commun. 2008; 377: 1195-1198
        • Van Lenten B.J.
        • Navab M.
        • Shih D.
        • et al.
        The role of high-density lipoproteins in oxidation and inflammation.
        Trends Cardiovasc Med. 2001; 11: 155-161
        • Cai J.
        • Kang Z.
        • Liu K.
        • et al.
        Neuroprotective effects of hydrogen saline in neonatal hypoxia-ischemia rat model.
        Brain Res. 2009; 1256: 129-137
        • Itahara T.
        • Suehiro T.
        • Ikeda Y.
        • et al.
        Serum paraoxonase and arylesterase activities in hemodialysis patients.
        J Atheroscler Thromb. 2000; 7: 152-158
        • Ayaori M.
        • Sawada S.
        • Yonemura A.
        • et al.
        Glucocorticoid receptor regulates ATP-binding cassette transporter-A1 expression and apolipoprotein-mediated cholesterol efflux from macrophages.
        Arterioscler Thromb Vasc Biol. 2006; 26: 163-168
        • Jiang X.C.
        • Bruce C.
        • Mar J.
        • et al.
        Targeted mutation of plasma phospholipid transfer protein gene markedly reduces high-density lipoprotein levels.
        J Clin Invest. 1999; 103: 907-914
        • Smith J.D.
        • Miyata M.
        • Ginsberg M.
        • et al.
        Cyclic AMP induces apolipoprotein E binding activity and promotes cholesterol efflux from a macrophage cell line to apolipoprotein acceptors.
        J Biol Chem. 1996; 271: 30647-30655
        • Basu S.K.
        • Brown M.S.
        • Ho Y.K.
        • et al.
        Mouse macrophages synthesize and secrete a protein resembling apolipoprotein E.
        Proc Natl Acad Sci USA. 1981; 78: 7545-7549
        • Rikitake Y.
        • Hirata K.
        • Kawashima S.
        • et al.
        Inhibitory effect of inducible type nitric oxide synthase on oxidative modification of low density lipoprotein by vascular smooth muscle cells.
        Atherosclerosis. 1998; 136: 51-57
        • Weber C.
        • Zernecke A.
        • Libby P.
        The multifaceted contributions of leukocyte subsets to atherosclerosis: lessons from mouse models.
        Nat Rev Immunol. 2008; 8: 802-815
        • Iiyama K.
        • Hajra L.
        • Iiyama M.
        • et al.
        Patterns of vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 expression in rabbit and mouse atherosclerotic lesions and at sites predisposed to lesion formation.
        Circ Res. 1999; 85: 199-207
        • Khovidhunkit W.
        • Kim M.S.
        • Memon R.A.
        • et al.
        Effects of infection and inflammation on lipid and lipoprotein metabolism: mechanisms and consequences to the host.
        J Lipid Res. 2004; 45: 1169-1196
        • Esteve E.
        • Ricart W.
        • Fernandez-Real J.M.
        Dyslipidemia and inflammation: an evolutionary conserved mechanism.
        Clin Nutr. 2005; 24: 16-31
        • Kolovou G.
        • Anagnostopoulou K.
        • Mikhailidis D.P.
        • et al.
        Apolipoprotein E knockout models.
        Curr Pharm Des. 2008; 14: 338-351
        • LaRosa J.C.
        • He J.
        • Vupputuri S.
        Effect of statins on risk of coronary disease: a meta-analysis of randomized controlled trials.
        JAMA. 1999; 282: 2340-2346
        • Garg R.
        • Vasamreddy C.R.
        • Blumenthal R.S.
        Non-high-density lipoprotein cholesterol: why lower is better.
        Prev Cardiol. 2005; 8: 173-177
      1. Third report of the national cholesterol education program (NCEP) expert panel on detection, evaluation, and treatment of high blood cholesterol in adults (adult treatment panel III) final report.
        Circulation. 2002; 106: 3143-3421
        • Grundy S.M.
        Low-density lipoprotein, non-high-density lipoprotein, and apolipoprotein B as targets of lipid-lowering therapy.
        Circulation. 2002; 106: 2526-2529
        • Fukuda K.
        • Asoh S.
        • Ishikawa M.
        • et al.
        Inhalation of hydrogen gas suppresses hepatic injury caused by ischemia/reperfusion through reducing oxidative stress.
        Biochem Biophys Res Commun. 2007; 361: 670-674
        • White C.R.
        • Brock T.A.
        • Chang L.Y.
        • et al.
        Superoxide and peroxynitrite in atherosclerosis.
        Proc Natl Acad Sci USA. 1994; 91: 1044-1048
        • Hansson G.K.
        Immunological control mechanisms in plaque formation.
        Basic Res Cardiol. 1994; 89: 41-46
        • Li H.
        • Cybulsky M.I.
        • Gimbrone Jr., M.A.
        • et al.
        Inducible expression of vascular cell adhesion molecule-1 by vascular smooth muscle cells in vitro and within rabbit atheroma.
        Am J Pathol. 1993; 143: 1551-1559
        • van Leeuwen M.
        • Gijbels M.J.
        • Duijvestijn A.
        • et al.
        Accumulation of myeloperoxidase-positive neutrophils in atherosclerotic lesions in LDLR−/− mice.
        Arterioscler Thromb Vasc Biol. 2008; 28: 84-89
        • Zerfaoui M.
        • Suzuki Y.
        • Naura A.S.
        • et al.
        Nuclear translocation of p65 NF-kappaB is sufficient for VCAM-1, but not ICAM-1, expression in TNF-stimulated smooth muscle cells: differential requirement for PARP-1 expression and interaction.
        Cell Signal. 2008; 20: 186-194
        • Guo L.
        • Song Z.
        • Li M.
        • et al.
        Scavenger receptor BI protects against septic death through its role in modulating inflammatory response.
        J Biol Chem. 2009; 284: 19826-19834
        • Krane V.
        • Drechsler C.
        • Wanner C.
        Dyslipidemia, inflammation and dialysis outcomes: what we know now.
        Curr Opin Nephrol Hypertens. 2006; 15: 566-570
        • Lim W.S
        • Timmins J.M.
        • Seimon T.A.
        • et al.
        Signal transducer and activator of transcription-1 is critical for apoptosis in macrophages subjected to endoplasmic reticulum stress in vitro and in advanced atherosclerotic lesions in vivo.
        Circulation. 2008; 117: 940-951
        • Ross R.
        Atherosclerosis – an inflammatory disease.
        N Engl J Med. 1999; 340: 115-126
        • Zernecke A.
        • Weber C.
        Inflammatory mediators in atherosclerotic vascular disease.
        Basic Res Cardiol. 2005; 100: 93-101
        • Wang N.
        • Silver D.L.
        • Thiele C.
        • et al.
        ATP-binding cassette transporter A1 (ABCA1) functions as a cholesterol efflux regulatory protein.
        J Biol Chem. 2001; 276: 23742-23747
        • Kennedy M.A.
        • Barrera G.C.
        • Nakamura K.
        • et al.
        ABCG1 has a critical role in mediating cholesterol efflux to HDL and preventing cellular lipid accumulation.
        Cell Metab. 2005; 1: 121-131
        • Wang N.
        • Lan D.
        • Chen W.
        • et al.
        ATP-binding cassette transporters G1 and G4 mediate cellular cholesterol efflux to high-density lipoproteins.
        Proc Natl Acad Sci USA. 2004; 101: 9774-9779
        • Ji Y.
        • Jian B.
        • Wang N.
        • et al.
        Scavenger receptor BI promotes high density lipoprotein-mediated cellular cholesterol efflux.
        J Biol Chem. 1997; 272: 20982-20985
        • Gordon T.
        • Castelli W.P.
        • Hjortland M.C.
        • et al.
        High density lipoprotein as a protective factor against coronary heart disease. The Framingham Study.
        Am J Med. 1977; 62: 707-714
        • Suzuki Y.
        • Sano M.
        • Hayashida K.
        • et al.
        Are the effects of alpha-glucosidase inhibitors on cardiovascular events related to elevated levels of hydrogen gas in the gastrointestinal tract?.
        FEBS Lett. 2009; 583: 2157-2159
        • Zhang S.H.
        • Reddick R.L.
        • Piedrahita J.A.
        • et al.
        Spontaneous hypercholesterolemia and arterial lesions in mice lacking apolipoprotein E.
        Science. 1992; 258: 468-471
        • Kris-Etherton P.M.
        • Dietschy J.
        Design criteria for studies examining individual fatty acid effects on cardiovascular disease risk factors: human and animal studies.
        Am J Clin Nutr. 1997; 65: 1590S-1596S