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Exogenous in vivo NO-donor treatment preserves p53 levels and protects vascular cells from apoptosis

  • Author Footnotes
    1 Both authors have equally contributed to the work.
    Xavier Duran
    Footnotes
    1 Both authors have equally contributed to the work.
    Affiliations
    Cardiovascular Research Center, CSIC-ICCC, Barcelona, Spain
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  • Author Footnotes
    1 Both authors have equally contributed to the work.
    Gemma Vilahur
    Footnotes
    1 Both authors have equally contributed to the work.
    Affiliations
    Cardiovascular Research Center, CSIC-ICCC, Barcelona, Spain

    CIBEROBN CB06/03, Instituto de Salud Carlos III, Spain
    Search for articles by this author
  • Lina Badimon
    Correspondence
    Corresponding author at: Cardiovascular Research Center, C/Sant Antoni Ma Claret 167, 08025 Barcelona, Spain. Tel.: +34 93 556 59 00; fax: +34 93 556 55 59.
    Affiliations
    Cardiovascular Research Center, CSIC-ICCC, Barcelona, Spain

    CIBEROBN CB06/03, Instituto de Salud Carlos III, Spain

    Cátedra Investigación Cardiovascular (UAB), Catalana-Occidente, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain
    Search for articles by this author
  • Author Footnotes
    1 Both authors have equally contributed to the work.

      Abstract

      Objective

      Nitric oxide (NO) is critical in cardiovascular protection. However, NO production is impaired in atherosclerosis resulting in thrombosis, vasoconstriction, and restenosis. Exogenous NO-donors (NOd) have proven protection against ischemia and thrombosis. However, their effect on vascular cell function remains unknown. Our objective was to determine the effect of NOd on vascular smooth muscle cell (VSMC) survival.

      Methods

      Pigs (N = 16) were randomly distributed in the following treatment groups: (I) LA419 (nitratethiol; 10-day p.o. 0.9 mg/kg bid); (II) LA816 (nitrosothiol; 2 h i.v. infusion, 6.6 nmol/(kg min)); (III) nitroglycerine (GTN, 2 h i.v. infusion, 2.5 mg/kg); and (IV) placebo-control. After sacrifice, pig aortic arch explants were either frozen or incubated with 20% homologous pig serum for different time periods (18 h, 2 and 10 days). Bcl2/bax ratio, phosphorylated-p53, cyclin-D2, casp-3, casp-8, and bax mRNA levels and protein expression were evaluated by real-time PCR and western blot, respectively.

      Results

      Within the first 2 days explants from NOd-treated animals had an increased pro-survival Bcl2/bax ratio compared with placebo-control (5× higher in LA816 and GTN i.v.-treated animals and 10× higher in 10-day orally treated LA419 animals; P < 0.05). Phosphorylated-P53-protein levels were consistently increased by all NOd-treatments vs. placebo-control (P < 0.05) and p53 mRNA levels showed an increase in placebo-controls at day 10 while they did not rise in NOd-treated animals (P < 0.05). Cyclin-D2, casp-3, and bax mRNA content were 6× lower in NOd-treated animals than in control explants at day-10 (P < 0.001). No changes were detected on caspase-8.

      Conclusions

      Taken together, our findings suggest that exogenous in vivo NO treatment seems to preserve VSMC from mitochondrial-dependent apoptosis and drive cells to quiescence through p53 increase.

      Keywords

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      References

        • Chesebro J.
        • Lam J.
        • Badimon L.
        • Fuster V.
        Restenosis after arterial angioplasty: a hemorrheologic response to injury.
        Am J Cardiol. 1987; 60: 10B-16B
        • Angelini G.D.
        • Newby A.C.
        The future of saphenous vein as a coronary artery bypass conduit.
        Eur Heart J. 1989; 10: 273-280
        • Vousden K.H.
        Activation of the p53 tumor suppressor protein.
        Biochim Biophys Acta. 2002; 1602: 47-59
        • Villalobo A.
        Nitric oxide and cell proliferation.
        FEBS. 2006; 273: 2329-2344
        • Rodriguez-Campos A.
        • Ruiz-Enriquez P.
        • Faraudo S.
        • Badimon L.
        Mitogen-induced p53 downregulation precedes vascular smooth muscle cell migration from healthy tunica media and proliferation.
        Arterioscler Thromb Vasc Biol. 2001; 2: 214-219
        • Brüne B.
        • von Knethen A.
        • Sandau K.
        Transcription factors p53 and HIF-1alpha as targets of nitric oxide.
        Cell Signal. 2001; 13: 525-533
        • Auchon D.
        • Jiang X.
        • Morgan D.
        • et al.
        Three-dimensional structure of the apoptosome: implications for assembly, procaspase-9 binding, and activation.
        Mol Cell. 2002; 9: 423-432
        • Walford G.
        • Loscalzo J.
        Nitric oxide in vascular biology.
        J Thromb Haemost. 2003; 1: 2112-2118
        • Jeremy J.Y.
        • Rowe D.
        • Emsley A.
        • Newby A.
        Nitric oxide and the proliferation of vascular smooth muscle cells.
        Cardiovasc Res. 1999; 43: 580-594
        • Sarkar R.
        • Gordon D.
        • Stanley J.
        • Webb R.
        Cell cycle effects of nitric oxide on vascular smooth muscle cells.
        Am J Physiol. 1997; 272: H1810-H1818
        • Tanner F.
        • Meier P.
        • Greutert H.
        • et al.
        Nitric oxide modulates expression of cell cycle regulatory proteins: a cytostatic strategy for inhibition of human vascular smooth muscle cell proliferation.
        Circulation. 2000; 101: 1982-1989
        • Messmer U.
        • Brüne B.
        Nitric oxide-induced apoptosis: p53-dependent and p53-independent signaling pathways.
        Biochem J. 1996; 319: 299-305
        • Jeremy J.Y.
        • Rowe D.
        • Emsley A.M.
        • Newby A.C.
        Nitric oxide and the proliferation of vascular smooth muscle cells.
        Cardiovasc Res. 1999; 43: 580-594
        • Vilahur G.
        • Segales E.
        • Casani L.
        • Badimon L.
        A novel anti-ischemic nitric oxide donor inhibits thrombosis without modifying haemodynamic parameters.
        Thromb Haemost. 2004; 91: 1035-1043
        • Urooj Zafar M.
        • Vilahur G.
        • Choi B.G.
        • et al.
        A novel anti-ischemic nitric oxide donor (LA419) reduces thrombogenesis in healthy human subjects.
        J Thromb Haemost. 2007;
        • Vilahur G.
        • Pena E.
        • Padro T.
        • Badimon L.
        Protein disulphide isomerase-mediated LA419-NO release provides additional antithrombotic effects to the blockade of the ADP receptor.
        Thromb Haemost. 2007; 97: 650-657
        • Vilahur G.
        • Segales E.
        • Salas E.
        • Badimon L.
        Effects of a novel platelet nitric oxide donor (LA816), aspirin, clopidogrel, and combined therapy in inhibiting flow- and lesion-dependent thrombosis in the porcine ex vivo model.
        Circulation. 2004; 110: 1686-1693
        • Manabe T.
        • Yamamoto A.
        • Satoh K.
        • Ichihara K.
        Tolerance to nitroglycerin induced by isosorbide-5-mononitrate infusion in vivo.
        Biol Pharm Bull. 2001; 24: 1370-1372
        • Loscalzo J.
        Nitric oxide insufficiency, platelet activation, and arterial thrombosis.
        Circ Res. 2001; 88: 756-762
        • Garg U.C.
        • Hassid A.
        Nitric oxide-generating vasodilators and 8-bromo-cyclic guanosine monophosphate inhibit mitogenesis and proliferation of cultured rat vascular smooth muscle cells.
        J Clin Invest. 1989; 83: 1774-1777
        • Geng Y.J.
        • Libby P.
        Progression of atheroma: a struggle between death and procreation.
        Arterioscler Thromb Vasc Biol. 2002; 22: 1370-1380
        • Ihling C.
        • Menzel G.
        • Wellens E.
        • Monting J.S.
        • Schaefer H.E.
        • Zeiher A.M.
        Topographical association between the cyclin-dependent kinases inhibitor P21, p53 accumulation, and cellular proliferation in human atherosclerotic tissue.
        Arterioscler Thromb Vasc Biol. 1997; 17: 2218-2224
        • Bennett M.R.
        • Littlewood T.D.
        • Schwartz S.M.
        • Weissberg P.L.
        Increased sensitivity of human vascular smooth muscle cells from atherosclerotic plaques to p53-mediated apoptosis.
        Circ Res. 1997; 81: 591-599
        • Dimmeler S.
        • Haendeler J.
        • Nehls M.
        • Zeiher A.M.
        Suppression of apoptosis by nitric oxide via inhibition of interleukin-1beta-converting enzyme (ICE)-like and cysteine protease protein (CPP)-32-like proteases.
        J Exp Med. 1997; 185: 601-607
        • Pollman M.J.
        • Yamada T.
        • Horiuchi M.
        • Gibbons G.H.
        Vasoactive substances regulate vascular smooth muscle cell apoptosis. Countervailing influences of nitric oxide and angiotensin II.
        Circ Res. 1996; 79: 748-756
        • Sancar A.
        • Lindsey-Boltz L.A.
        • Unsal-Kacmaz K.
        • Linn S.
        Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints.
        Annu Rev Biochem. 2004; 73: 39-85
        • Hengartner M.O.
        The biochemistry of apoptosis.
        Nature. 2000; 407: 770-776
        • Xie K.
        • Huang S.
        • Wang Y.
        • et al.
        Bcl-2 protects cells from cytokine-induced nitric-oxide-dependent apoptosis.
        Cancer Immunol Immunother. 1996; 43: 109-115
        • Green D.R.
        • Reed J.C.
        Mitochondria and apoptosis.
        Science. 1998; 281: 1309-1312
        • Janssens S.
        • Flaherty D.
        • Nong Z.
        • et al.
        Human endothelial nitric oxide synthase gene transfer inhibits vascular smooth muscle cell proliferation and neointima formation after balloon injury in rats.
        Circulation. 1998; 97: 1274-1281