Serum-induced proliferation of rabbit aortic smooth muscle cells from the contractile state is inhibited by 8-Br-CAMP but not 8-Br-cGMP

  • Kay Southgate
    Correspondence to: Kay M. Southgate, Department of Cardiology, University of Wales College of Medicine, Heath Park, Cardiff CF4 4XN, U.K. Tel.: 0222 755944 ext.: 2338.
    Department of Cardiology, University of Wales College of Medicine, Heath Park, Cardiff CF4 4XN UK
    Search for articles by this author
  • Andrew C. Newby
    Department of Cardiology, University of Wales College of Medicine, Heath Park, Cardiff CF4 4XN UK
    Search for articles by this author
      This paper is only available as a PDF. To read, Please Download here.


      A method is described for the quantification of vascular smooth muscle cell growth from individual explants of contractile rabbit aortic tunica media. The precision of the method probably depends on regular explant geometry (1-mm squares) and pooling sufficient explants. Serum-induced growth was quantified by measurements of ATP concentration, incorporation of [3H]thymidine and DNA concentration. The possible effects of endogenous vasodilator agents on growth were investigated by using lipid soluble analogues of their second messengers, namely 8-Br-cAMP and 8-Br-cGMP, which are known to relax rabbit aortic strips. Cell growth was inhibited concentration-dependently by 8-Br-cAMP but not 8-Br-cGMP (0.01-1 mM). The effect of 8-Br-cAMP was reversible, and also occurred when addition was delayed until after growth had commenced. The results imply that endogenous vasodilators such as prostacyclin, adenosine and adrenaline, which increase cAMP concentration, may normally suppress smooth muscle cell growth, whereas nitric oxide and atriopeptins, which increase cGMP concentration, may not.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Ross R.
        • Glomset J.A.
        The pathogenesis of atherosclerosis (first of two parts).
        N. Engl. J. Med. 1976; 295: 369
        • Chesebro J.H.
        • Lam J.Y.T.
        • Badimon L.
        • Fuster V.
        Restenosis after arterial angioplasty: a haemorrheologic response to injury.
        Am. J. Cardiol. 1987; 60: 10B
        • Angelini G.D.
        • Newby A.C.
        The future of saphenous vein as a coronary artery bypass conduit.
        Eur. Heart J. 1989; 10: 273
        • Campbell J.H.
        • Campbell G.R.
        Endothelial cell influences on vascular smooth muscle cell phenotype.
        Annu. Rev. Physiol. 1986; 48: 295
        • Campbell J.H.
        • Black M.J.
        • Campbell G.R.
        Replication of smooth muscle cells in atherosclerosis and hypertension.
        in: Meyer P. Marche P. Blood Cells and Arteries in Hypertension. Raven Press, New York1989: 15
        • Schwartz S.M.
        • Campbell G.R.
        • Campbell J.H.
        Replication of smooth muscle cells in vascular disease.
        Circ. Res. 1986; 58: 427
        • Heyns A.
        • Eldor A.
        • Vlodavsky I.
        • Kaiser N.
        • Fridman R.
        • Panet A.
        The antiproliferative effect of Interferon and the mitogenic activity of growth factors are independent cell cycle events.
        Exp. Cell Res. 1985; 161: 297
        • Assoian R.K.
        • Sporn M.B.
        Type B transforming growth factor in human platelets: release during platelet degranulation and action on vascular smooth muscle cells.
        J. Cell. Biol. 1986; 102: 1217
        • Owen N.E.
        Prostacyclin can inhibit DNA synthesis in vascular smooth muscle cells.
        in: Bailey J.M.B. Prostaglandins, Leukotrienes, and Lipoxins. Plenum Publishing Corporation, New York1985: 193 (Ch. 19)
        • Loesberg C.
        • Van Wijk R.
        • Zandbergen J.
        • Van Aken W.G.
        • Van Mourik J.A.
        • De Groot P.H.G.
        Cell cycle dependent inhibition of human vascular smooth muscle cell proliferation by prostaglandin E1.
        Exp. Cell Res. 1985; 160: 117
        • Jonzon B.
        • Nilsson J.
        • Fredholm B.B.
        Adenosine receptor-mediated changes in cyclic AMP production and DNA synthesis in cultured arterial smooth muscle cells.
        J. Cell Physiol. 1985; 124: 451
        • 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
        • Abell T.J.
        • Richards A.M.
        • Ikram H.
        • Espinier E.A.
        • Yandle T.
        Atrial natriuretic factor inhibits proliferation of vascular smooth muscle cells stimulated by platelet-derived growth factor.
        Biochem. Biophys. Res. Commun. 1989; 60: 1392
        • Khym J.X.
        An analytical system for rapid separation of tissue nucleotides at low pressures on conventional anion exchangers.
        Clin. Chem. 1975; 21/9: 1245
        • Kissane J.M.
        • Robins E.
        The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system.
        J. Biol. Chem. 1958; 233: 184
        • Spielmann H.
        • Jacob-Müller U.
        • Schulz P.
        Simple assay of 0.1–1.0 pmol of ATP, ADP and AMP in single somatic cells using purified luciferin luciferase.
        Anal. Biochem. 1981; 113: 172
        • Needleman P.
        • Blehm D.J.
        Effect of epinephrine and potassium choride on contraction and energy intermediates in rabbit thoracic aorta strips.
        Life Sci. 1970; 9: 1181
        • Angus J.A.
        • Cocks T.M.
        Endothelium-derived relaxing factor.
        Phatmac. Ther. 1989; 41: 303
        • Collins P.
        • Griffith T.M.
        • Henderson A.H.
        • Lewis M.J.
        Endothelium-derived relaxing factor alters calcium fluxes in rabbit aorta: a cyclic guanosine monophosphatemediated effect.
        J. Physiol. 1986; 381: 427
        • Martin W.
        • Morgan R.O.
        • Smith J.A.
        • White D.C.
        Atriopeptin II-induced relaxation of rabbit aorta is potentiaced by M&B 22,948 but not blocked by haemoglobin.
        Br. J. Pharmacol. 1986; 89: 557
        • Collins P.
        • Henderson A.H.
        • Lang D.
        • Lewis M.J.
        Endothelium-derived relaxing factor and nitroprusside compared in noradrenaline and K+-contracted rabbit and rat aortae.
        J. Physiol. 1988; 400: 395
        • Lang D.
        • Lewis M.J.
        Endothelium-derived relaxing factor inhibits the formation of inositol triphosphate by rabbit aorta.
        J. Physiol. 1989; 411: 45
        • Jarmolych J.
        • Daoud A.S.
        • Landau J.
        • Fritz K.E.
        • McElvene E.
        Aortic media explants: cell proliferation and production of mucopolysacchrides, collagen and elastic tissue.
        Exp. Mol. Pathol. 1968; 9: 171
        • Angelini G.D.
        • Passani S.L.
        • Breckenridge I.M.
        • Newby A.C.
        The nature and pressure-dependence of damage induced by distension of human saphenous vein coronary artery bypass grafts.
        Cardiovasc. Res. 1987; 21: 902
        • McGilvery R.W.
        • Murray T.W.
        Calculated equilibria of phosphocreatine and adenosine phosphate during utilization of high energy phosphate by muscle.
        J. Biol. Chem. 1974; 249: 5845
        • Beebe S.J.
        • Holloway R.
        • Rannels S.R.
        • Corbin J.D.
        Two classes of cAMP analogs which are selective for the two different cGMP-binding sites of type II protein kinase demonstrate synergism when added together to intact adipocytes.
        J. Biol. Chem. 1984; 259: 3539
        • Orekhov A.N.
        • Tertov V.V.
        • Kudryashov S.A.
        • Khashimov K.A.
        • Smirnov V.N.
        Primary culture of human aortic intima cells as a model for testing antiatherosclerotic drugs.
        Atheiosclerosis. 1986; 60: 101
        • Magnaldo I.
        • Pouyssegur J.
        • Paris S.
        Cyclic AMP inhibits mitogen-induced DNA synthesis in hamster fibroblasts, regardless of the signalling pathway involved.
        FEBS Lett. 1985; 245: 65
        • Corbin J.D.
        • Øgried D.
        • Miller J.P.
        • Suva R.H.
        • Jastorff B.
        • Døskeland S.O.
        Studies of cGMP analog specificity and function of the two intrasubunit binding sites of cGMP-dependent protein kinase.
        J. Biol. Chem. 1986; 261: 1208
        • Resink T.J.
        • Scott-Burden T.
        • Baur U.
        • Jones C.R.
        • Bühler F.R.
        Atrial natriuretic peptide induces breakdown of phosphatidylinositol phosphates in cultured vascular smooth-muscle cells.
        Eur. J. Biochem. 1988; 172: 499
        • Radomski M.W.
        • Palmer R.M.J.
        • Moncada S.
        The anti-aggregating properties of vascular endothelium: interactions between prostacyclin and nitric oxide.
        Br. J. Pharmacol. 1987; 92: 639
        • Morgan J.P.
        • Morgan K.G.
        Alteration of cytoplasmic ionized calcium levels in smooth muscle by vasodilators in the ferret.
        J. Physiol. 1984; 357: 539
        • Brunwald J.
        • Haudenschild C.C.
        Intimal injury in vivo activates vascular smooth muscle cell migration and explant outgrowth in vitro.
        Arteriosclerosis. 1984; 4: 183
        • McMurray H.F.
        Macrophage growth factors in atherogenesis.
        in: 3rd edn. PhD Thesis. University of Cambridge, Cambridge1989