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The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1985 Aug;76(2):703–708. doi: 10.1172/JCI112024

N-Acetylcysteine potentiates inhibition of platelet aggregation by nitroglycerin.

J Loscalzo
PMCID: PMC423881  PMID: 2863286

Abstract

Platelet aggregation is currently felt to play an important role in the pathogenesis of ischemic vascular disorders. The smooth muscle relaxant, nitroglycerin, has been shown to inhibit platelet aggregation in vitro, but at concentrations that were felt to be unattainable in vivo. Because the in vivo action of nitroglycerin on smooth muscle cells has been shown to depend on the presence of reduced cytosolic sulfhydryl groups, the inhibitory effect of nitroglycerin on platelet aggregation was examined in the presence of the reduced thiol, N-acetylcysteine. Millimolar concentrations of N-acetylcysteine potentiated markedly the inhibitory effect of nitroglycerin on platelet aggregation induced by ADP, epinephrine, collagen, and arachidonate, decreasing the 50% inhibitory concentration (IC50) approximately 50-fold for each of these agents. Other guanylate cyclase activators inhibited ADP-induced aggregation similarly and this inhibition was likewise potentiated by N-acetylcysteine. Platelet guanosine 3',5'-cyclic monophosphate content increased fivefold in the presence of nitroglycerin and N-acetylcysteine 2 min before maximal inhibition of ADP-induced aggregation was achieved, while simultaneously measured cyclic AMP did not change relative to base-line levels. In the absence of N-acetylcysteine, nitroglycerin induced a marked decrease in platelet-reduced glutathione content as S-nitroso-thiol adducts were produced. The synthetic S-nitroso-thiol, S-nitroso-N-acetylcysteine, markedly inhibited platelet aggregation with an IC50 of 6 nM. These data show that N-acetylcysteine markedly potentiates the inhibition of platelet aggregation by nitroglycerin and likely does so by inducing the formation of an S-nitrosothiol adduct(s), which activate guanylate cyclase.

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Selected References

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  1. Akkerman J. W. Regulation of carbohydrate metabolism in platelets. A review. Thromb Haemost. 1978 Jun 30;39(3):712–724. [PubMed] [Google Scholar]
  2. BEUTLER E., DURON O., KELLY B. M. Improved method for the determination of blood glutathione. J Lab Clin Med. 1963 May;61:882–888. [PubMed] [Google Scholar]
  3. BORN G. V., CROSS M. J. THE AGGREGATION OF BLOOD PLATELETS. J Physiol. 1963 Aug;168:178–195. doi: 10.1113/jphysiol.1963.sp007185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chiang T. M., Beachey E. H., Kang A. H. Interaction of a chick skin collagen fragment (alpha1-CB5) with human platelets. Biochemical studies during the aggregation and release reaction. J Biol Chem. 1975 Sep 10;250(17):6916–6922. [PubMed] [Google Scholar]
  5. Chiang T. M., Dixit S. N., Kang A. H. Effect of cyclic 3',5'-guanosine monophosphate on human platelet function. J Lab Clin Med. 1976 Aug;88(2):215–221. [PubMed] [Google Scholar]
  6. Claesson H. E., Malmsten C. On the interrelationship of prostaglandin endoperoxide G2 and cyclic nucleotides in platelet function. Eur J Biochem. 1977 Jun 1;76(1):277–284. doi: 10.1111/j.1432-1033.1977.tb11593.x. [DOI] [PubMed] [Google Scholar]
  7. Collier H. B. Letter: A note on the molar absorptivity of reduced Ellman's reagent, 3-carboxylato-4-nitrothiophenolate. Anal Biochem. 1973 Nov;56(1):310–311. doi: 10.1016/0003-2697(73)90196-6. [DOI] [PubMed] [Google Scholar]
  8. ELLMAN G. L. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959 May;82(1):70–77. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
  9. Ellis E. F., Oelz O., Roberts L. J., 2nd, Payne N. A., Sweetman B. J., Nies A. S., Oates J. A. Coronary arterial smooth muscle contraction by a substance released from platelets: evidence that it is thromboxane A2. Science. 1976 Sep 17;193(4258):1135–1137. doi: 10.1126/science.959827. [DOI] [PubMed] [Google Scholar]
  10. Feldman R. L., Conti C. R. Relief of myocardial ischemia with nitroglycerin: what is the mechanism? Circulation. 1981 Dec;64(6):1098–1100. doi: 10.1161/01.cir.64.6.1098. [DOI] [PubMed] [Google Scholar]
  11. Feldman R. L., Pepine C. J., Conti C. R. Magnitude of dilatation of large and small coronary arteries of nitroglycerin. Circulation. 1981 Aug;64(2):324–333. doi: 10.1161/01.cir.64.2.324. [DOI] [PubMed] [Google Scholar]
  12. Glass D. B., Gerrard J. M., Townsend D., Carr D. W., White J. G., Goldberg N. D. The involvement of prostaglandin endoperoxide formation in the elevation of cyclic GMP levels during platelet aggregation. J Cyclic Nucleotide Res. 1977 Feb;3(1):37–44. [PubMed] [Google Scholar]
  13. Greenwald J. E., Bianchine J. R., Wong L. K. The production of the arachidonate metabolite HETE in vascular tissue. Nature. 1979 Oct 18;281(5732):588–589. doi: 10.1038/281588a0. [DOI] [PubMed] [Google Scholar]
  14. Gruetter C. A., Gruetter D. Y., Lyon J. E., Kadowitz P. J., Ignarro L. J. Relationship between cyclic guanosine 3':5'-monophosphate formation and relaxation of coronary arterial smooth muscle by glyceryl trinitrate, nitroprusside, nitrite and nitric oxide: effects of methylene blue and methemoglobin. J Pharmacol Exp Ther. 1981 Oct;219(1):181–186. [PubMed] [Google Scholar]
  15. Habeeb A. F. Chemical evaluation of conformational differences in native and chemically modified proteins. Biochim Biophys Acta. 1966 Feb 28;115(2):440–454. doi: 10.1016/0304-4165(66)90442-9. [DOI] [PubMed] [Google Scholar]
  16. Habig W. H., Keen J. H., Jakoby W. B. Glutathione S-transferase in the formation of cyanide from organic thiocyantes and as an organic nitrate reductase. Biochem Biophys Res Commun. 1975 May 19;64(2):501–506. doi: 10.1016/0006-291x(75)90349-6. [DOI] [PubMed] [Google Scholar]
  17. Haerem J. W. Mural platelet microthrombi and major acute lesions of main epicardial arteries in sudden coronary death. Atherosclerosis. 1974 May-Jun;19(3):529–541. doi: 10.1016/s0021-9150(74)80017-1. [DOI] [PubMed] [Google Scholar]
  18. Haerem J. W. Platelet aggregates in intramyocardial vessels of patients dying suddenly and unexpectedly of coronary artery disease. Atherosclerosis. 1972 Mar-Apr;15(2):199–213. doi: 10.1016/0021-9150(72)90070-6. [DOI] [PubMed] [Google Scholar]
  19. Hampton J. R., Harrison M. J., Honour A. J., Mitchell J. R. Platelet behaviour and drugs used in cardiovascular disease. Cardiovasc Res. 1967 Apr;1(2):101–107. doi: 10.1093/cvr/1.2.101. [DOI] [PubMed] [Google Scholar]
  20. Horowitz J. D., Antman E. M., Lorell B. H., Barry W. H., Smith T. W. Potentiation of the cardiovascular effects of nitroglycerin by N-acetylcysteine. Circulation. 1983 Dec;68(6):1247–1253. doi: 10.1161/01.cir.68.6.1247. [DOI] [PubMed] [Google Scholar]
  21. Ignarro L. J., Gruetter C. A. Requirement of thiols for activation of coronary arterial guanylate cyclase by glyceryl trinitrate and sodium nitrite: possible involvement of S-nitrosothiols. Biochim Biophys Acta. 1980 Aug 13;631(2):221–231. doi: 10.1016/0304-4165(80)90297-4. [DOI] [PubMed] [Google Scholar]
  22. Ignarro L. J., Kadowitz P. J., Baricos W. H. Evidence that regulation of hepatic guanylate cyclase activity involves interactions between catalytic site -SH groups and both substrate and activator. Arch Biochem Biophys. 1981 Apr 15;208(1):75–86. doi: 10.1016/0003-9861(81)90125-9. [DOI] [PubMed] [Google Scholar]
  23. Ignarro L. J., Lippton H., Edwards J. C., Baricos W. H., Hyman A. L., Kadowitz P. J., Gruetter C. A. Mechanism of vascular smooth muscle relaxation by organic nitrates, nitrites, nitroprusside and nitric oxide: evidence for the involvement of S-nitrosothiols as active intermediates. J Pharmacol Exp Ther. 1981 Sep;218(3):739–749. [PubMed] [Google Scholar]
  24. Marcus A. J. The role of lipids in platelet function: with particular reference to the arachidonic acid pathway. J Lipid Res. 1978 Sep;19(7):793–826. [PubMed] [Google Scholar]
  25. McGregor M. Pathogenesis of angina pectoris and role of nitrates in relief of myocardial ischemia. Am J Med. 1983 Jun 27;74(6B):21–27. doi: 10.1016/0002-9343(83)90850-1. [DOI] [PubMed] [Google Scholar]
  26. Mellion B. T., Ignarro L. J., Myers C. B., Ohlstein E. H., Ballot B. A., Hyman A. L., Kadowitz P. J. Inhibition of human platelet aggregation by S-nitrosothiols. Heme-dependent activation of soluble guanylate cyclase and stimulation of cyclic GMP accumulation. Mol Pharmacol. 1983 May;23(3):653–664. [PubMed] [Google Scholar]
  27. Mellion B. T., Ignarro L. J., Ohlstein E. H., Pontecorvo E. G., Hyman A. L., Kadowitz P. J. Evidence for the inhibitory role of guanosine 3', 5'-monophosphate in ADP-induced human platelet aggregation in the presence of nitric oxide and related vasodilators. Blood. 1981 May;57(5):946–955. [PubMed] [Google Scholar]
  28. Needleman P., Jakschik B., Johnson E. M., Jr Sulfhydryl requirement for relaxation of vascular smooth muscle. J Pharmacol Exp Ther. 1973 Nov;187(2):324–331. [PubMed] [Google Scholar]
  29. Needleman P., Johnson E. M., Jr Mechanism of tolerance development to organic nitrates. J Pharmacol Exp Ther. 1973 Mar;184(3):709–715. [PubMed] [Google Scholar]
  30. Needleman P., Kulkarni P. S., Raz A. Coronary tone modulation: formation and actions of prostaglandins, endoperoxides, and thromboxanes. Science. 1977 Jan 28;195(4276):409–412. doi: 10.1126/science.831285. [DOI] [PubMed] [Google Scholar]
  31. Needleman P. Organic nitrate metabolism. Annu Rev Pharmacol Toxicol. 1976;16:81–93. doi: 10.1146/annurev.pa.16.040176.000501. [DOI] [PubMed] [Google Scholar]
  32. Schafer A. I., Alexander R. W., Handin R. I. Inhibition of platelet function by organic nitrate vasodilators. Blood. 1980 Apr;55(4):649–654. [PubMed] [Google Scholar]
  33. Synek P., Rysánek K., Spánková H., Mlejnková M. The effect of ethanol and nitroglycerin on platelet aggregation. Act Nerv Super (Praha) 1970 Jan 12;12(1):77–78. [PubMed] [Google Scholar]
  34. Warren S. E., Francis G. S. Nitroglycerin and nitrate esters. Am J Med. 1978 Jul;65(1):53–62. doi: 10.1016/0002-9343(78)90692-7. [DOI] [PubMed] [Google Scholar]
  35. Weiss A., Baenziger N. L., Atkinson J. P. Platelet release reaction and intracellular cGMP. Blood. 1978 Sep;52(3):524–531. [PubMed] [Google Scholar]

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