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J Clin Invest. Mar 15, 1998; 101(6): 1408–1414.
PMCID: PMC508696

L-2-Oxothiazolidine-4-carboxylic acid reverses endothelial dysfunction in patients with coronary artery disease.


The effective action of endothelium-derived nitric oxide (EDNO) is impaired in patients with atherosclerosis. This impairment has been attributed in part to increased vascular oxidative stress. EDNO action is improved by administration of ascorbic acid, a water-soluble antioxidant. Ascorbic acid is a potent free-radical scavenger in plasma, and also regulates intracellular redox state in part by sparing cellular glutathione. We specifically investigated the role of intracellular redox state in EDNO action by examining the effect of L-2-oxo-4-thiazolidine carboxylate (OTC) on EDNO-dependent, flow-mediated dilation in a randomized double-blind placebo-controlled study of patients with angiographically proven coronary artery disease. OTC augments intracellular glutathione by providing substrate cysteine for glutathione synthesis. Brachial artery flow-mediated dilation was examined with high-resolution ultrasound before and after oral administration of 4.5 g of OTC or placebo in 48 subjects with angiographically documented coronary artery disease. Placebo treatment produced no change in flow-mediated dilation (7.0+/-3.9% vs. 7.2+/-3.7%), whereas OTC treatment was associated with a significant improvement in flow-mediated dilation (6.6+/-4.4% vs. 11.0+/-6.3%; P = 0.005). OTC had no effect on arterial dilation to nitroglycerin, systemic blood pressure, heart rate, or reactive hyperemia. These data suggest that augmenting cellular glutathione levels improves EDNO action in human atherosclerosis. Cellular redox state may be an important regulator of EDNO action, and is a potential target for therapy in patients with coronary artery disease.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Vita JA, Treasure CB, Ganz P, Cox DA, Fish RD, Selwyn AP. Control of shear stress in the epicardial coronary arteries of humans: impairment by atherosclerosis. J Am Coll Cardiol. 1989 Nov 1;14(5):1193–1199. [PubMed]
  • Ludmer PL, Selwyn AP, Shook TL, Wayne RR, Mudge GH, Alexander RW, Ganz P. Paradoxical vasoconstriction induced by acetylcholine in atherosclerotic coronary arteries. N Engl J Med. 1986 Oct 23;315(17):1046–1051. [PubMed]
  • Vita JA, Treasure CB, Nabel EG, McLenachan JM, Fish RD, Yeung AC, Vekshtein VI, Selwyn AP, Ganz P. Coronary vasomotor response to acetylcholine relates to risk factors for coronary artery disease. Circulation. 1990 Feb;81(2):491–497. [PubMed]
  • Levine GN, Keaney JF, Jr, Vita JA. Cholesterol reduction in cardiovascular disease. Clinical benefits and possible mechanisms. N Engl J Med. 1995 Feb 23;332(8):512–521. [PubMed]
  • Keaney JF, Jr, Vita JA. Atherosclerosis, oxidative stress, and antioxidant protection in endothelium-derived relaxing factor action. Prog Cardiovasc Dis. 1995 Sep-Oct;38(2):129–154. [PubMed]
  • Ohara Y, Peterson TE, Harrison DG. Hypercholesterolemia increases endothelial superoxide anion production. J Clin Invest. 1993 Jun;91(6):2546–2551. [PMC free article] [PubMed]
  • Keaney JF, Jr, Xu A, Cunningham D, Jackson T, Frei B, Vita JA. Dietary probucol preserves endothelial function in cholesterol-fed rabbits by limiting vascular oxidative stress and superoxide generation. J Clin Invest. 1995 Jun;95(6):2520–2529. [PMC free article] [PubMed]
  • Rubanyi GM, Vanhoutte PM. Superoxide anions and hyperoxia inactivate endothelium-derived relaxing factor. Am J Physiol. 1986 May;250(5 Pt 2):H822–H827. [PubMed]
  • Gryglewski RJ, Palmer RM, Moncada S. Superoxide anion is involved in the breakdown of endothelium-derived vascular relaxing factor. Nature. 1986 Apr 3;320(6061):454–456. [PubMed]
  • Kugiyama K, Kerns SA, Morrisett JD, Roberts R, Henry PD. Impairment of endothelium-dependent arterial relaxation by lysolecithin in modified low-density lipoproteins. Nature. 1990 Mar 8;344(6262):160–162. [PubMed]
  • Chin JH, Azhar S, Hoffman BB. Inactivation of endothelial derived relaxing factor by oxidized lipoproteins. J Clin Invest. 1992 Jan;89(1):10–18. [PMC free article] [PubMed]
  • Mügge A, Elwell JH, Peterson TE, Hofmeyer TG, Heistad DD, Harrison DG. Chronic treatment with polyethylene-glycolated superoxide dismutase partially restores endothelium-dependent vascular relaxations in cholesterol-fed rabbits. Circ Res. 1991 Nov;69(5):1293–1300. [PubMed]
  • Keaney JF, Jr, Gaziano JM, Xu A, Frei B, Curran-Celentano J, Shwaery GT, Loscalzo J, Vita JA. Dietary antioxidants preserve endothelium-dependent vessel relaxation in cholesterol-fed rabbits. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11880–11884. [PMC free article] [PubMed]
  • Simon BC, Haudenschild CC, Cohen RA. Preservation of endothelium-dependent relaxation in atherosclerotic rabbit aorta by probucol. J Cardiovasc Pharmacol. 1993 Jun;21(6):893–901. [PubMed]
  • Anderson TJ, Meredith IT, Yeung AC, Frei B, Selwyn AP, Ganz P. The effect of cholesterol-lowering and antioxidant therapy on endothelium-dependent coronary vasomotion. N Engl J Med. 1995 Feb 23;332(8):488–493. [PubMed]
  • Meister A. Glutathione-ascorbic acid antioxidant system in animals. J Biol Chem. 1994 Apr 1;269(13):9397–9400. [PubMed]
  • Ting HH, Timimi FK, Boles KS, Creager SJ, Ganz P, Creager MA. Vitamin C improves endothelium-dependent vasodilation in patients with non-insulin-dependent diabetes mellitus. J Clin Invest. 1996 Jan 1;97(1):22–28. [PMC free article] [PubMed]
  • Heitzer T, Just H, Münzel T. Antioxidant vitamin C improves endothelial dysfunction in chronic smokers. Circulation. 1996 Jul 1;94(1):6–9. [PubMed]
  • Ting HH, Timimi FK, Haley EA, Roddy MA, Ganz P, Creager MA. Vitamin C improves endothelium-dependent vasodilation in forearm resistance vessels of humans with hypercholesterolemia. Circulation. 1997 Jun 17;95(12):2617–2622. [PubMed]
  • Levine GN, Frei B, Koulouris SN, Gerhard MD, Keaney JF, Jr, Vita JA. Ascorbic acid reverses endothelial vasomotor dysfunction in patients with coronary artery disease. Circulation. 1996 Mar 15;93(6):1107–1113. [PubMed]
  • Ma XL, Lopez BL, Liu GL, Christopher TA, Gao F, Guo Y, Feuerstein GZ, Ruffolo RR, Jr, Barone FC, Yue TL. Hypercholesterolemia impairs a detoxification mechanism against peroxynitrite and renders the vascular tissue more susceptible to oxidative injury. Circ Res. 1997 Jun;80(6):894–901. [PubMed]
  • Williamson JM, Meister A. Stimulation of hepatic glutathione formation by administration of L-2-oxothiazolidine-4-carboxylate, a 5-oxo-L-prolinase substrate. Proc Natl Acad Sci U S A. 1981 Feb;78(2):936–939. [PMC free article] [PubMed]
  • Williamson JM, Boettcher B, Meister A. Intracellular cysteine delivery system that protects against toxicity by promoting glutathione synthesis. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6246–6249. [PMC free article] [PubMed]
  • Boesgaard S, Poulsen HE, Aldershvile J, Loft S, Anderson ME, Meister A. Acute effects of nitroglycerin depend on both plasma and intracellular sulfhydryl compound levels in vivo. Effect of agents with different sulfhydryl-modulating properties. Circulation. 1993 Feb;87(2):547–553. [PubMed]
  • Boesgaard S, Aldershvile J, Poulsen HE, Loft S, Anderson ME, Meister A. Nitrate tolerance in vivo is not associated with depletion of arterial or venous thiol levels. Circ Res. 1994 Jan;74(1):115–120. [PubMed]
  • Porta P, Aebi S, Summer K, Lauterburg BH. L-2-oxothiazolidine-4-carboxylic acid, a cysteine prodrug: pharmacokinetics and effects on thiols in plasma and lymphocytes in human. J Pharmacol Exp Ther. 1991 Apr;257(1):331–334. [PubMed]
  • Hamsten A, Walldius G, Szamosi A, Dahlen G, de Faire U. Relationship of angiographically defined coronary artery disease to serum lipoproteins and apolipoproteins in young survivors of myocardial infarction. Circulation. 1986 Jun;73(6):1097–1110. [PubMed]
  • Lieberman EH, Gerhard MD, Uehata A, Walsh BW, Selwyn AP, Ganz P, Yeung AC, Creager MA. Estrogen improves endothelium-dependent, flow-mediated vasodilation in postmenopausal women. Ann Intern Med. 1994 Dec 15;121(12):936–941. [PubMed]
  • Lieberman EH, Gerhard MD, Uehata A, Selwyn AP, Ganz P, Yeung AC, Creager MA. Flow-induced vasodilation of the human brachial artery is impaired in patients <40 years of age with coronary artery disease. Am J Cardiol. 1996 Dec 1;78(11):1210–1214. [PubMed]
  • Assmann G, Schriewer H, Schmitz G, Hägele EO. Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem. 1983 Dec;29(12):2026–2030. [PubMed]
  • Friedewald WT, Levy RI, Fredrickson DS. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem. 1972 Jun;18(6):499–502. [PubMed]
  • Frei B, England L, Ames BN. Ascorbate is an outstanding antioxidant in human blood plasma. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6377–6381. [PMC free article] [PubMed]
  • Fiskerstrand T, Refsum H, Kvalheim G, Ueland PM. Homocysteine and other thiols in plasma and urine: automated determination and sample stability. Clin Chem. 1993 Feb;39(2):263–271. [PubMed]
  • Winkler BS. In vitro oxidation of ascorbic acid and its prevention by GSH. Biochim Biophys Acta. 1987 Sep 11;925(3):258–264. [PubMed]
  • Mårtensson J, Han J, Griffith OW, Meister A. Glutathione ester delays the onset of scurvy in ascorbate-deficient guinea pigs. Proc Natl Acad Sci U S A. 1993 Jan 1;90(1):317–321. [PMC free article] [PubMed]
  • Hecker M, Siegle I, Macarthur H, Sessa WC, Vane JR. Role of intracellular thiols in release of EDRF from cultured endothelial cells. Am J Physiol. 1992 Mar;262(3 Pt 2):H888–H896. [PubMed]
  • Murphy ME, Piper HM, Watanabe H, Sies H. Nitric oxide production by cultured aortic endothelial cells in response to thiol depletion and replenishment. J Biol Chem. 1991 Oct 15;266(29):19378–19383. [PubMed]
  • Mügge A, Elwell JH, Peterson TE, Harrison DG. Release of intact endothelium-derived relaxing factor depends on endothelial superoxide dismutase activity. Am J Physiol. 1991 Feb;260(2 Pt 1):C219–C225. [PubMed]
  • Ghigo D, Alessio P, Foco A, Bussolino F, Costamagna C, Heller R, Garbarino G, Pescarmona GP, Bosia A. Nitric oxide synthesis is impaired in glutathione-depleted human umbilical vein endothelial cells. Am J Physiol. 1993 Sep;265(3 Pt 1):C728–C732. [PubMed]
  • Komori Y, Hyun J, Chiang K, Fukuto JM. The role of thiols in the apparent activation of rat brain nitric oxide synthase (NOS). J Biochem. 1995 Apr;117(4):923–927. [PubMed]
  • Stuehr DJ, Kwon NS, Nathan CF. FAD and GSH participate in macrophage synthesis of nitric oxide. Biochem Biophys Res Commun. 1990 Apr 30;168(2):558–565. [PubMed]
  • Hobbs AJ, Fukuto JM, Ignarro LJ. Formation of free nitric oxide from l-arginine by nitric oxide synthase: direct enhancement of generation by superoxide dismutase. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10992–10996. [PMC free article] [PubMed]
  • Hofmann H, Schmidt HH. Thiol dependence of nitric oxide synthase. Biochemistry. 1995 Oct 17;34(41):13443–13452. [PubMed]
  • Howells DW, Hyland K. Direct analysis of tetrahydrobiopterin in cerebrospinal fluid by high-performance liquid chromatography with redox electrochemistry: prevention of autoxidation during storage and analysis. Clin Chim Acta. 1987 Jul 30;167(1):23–30. [PubMed]
  • Heales S, Hyland K. Determination of quinonoid dihydrobiopterin by high-performance liquid chromatography and electrochemical detection. J Chromatogr. 1989 Sep 29;494:77–85. [PubMed]
  • Patel JM, Abeles AJ, Block ER. Nitric oxide exposure and sulfhydryl modulation alter L-arginine transport in cultured pulmonary artery endothelial cells. Free Radic Biol Med. 1996;20(5):629–637. [PubMed]
  • Heinzel B, John M, Klatt P, Böhme E, Mayer B. Ca2+/calmodulin-dependent formation of hydrogen peroxide by brain nitric oxide synthase. Biochem J. 1992 Feb 1;281(Pt 3):627–630. [PMC free article] [PubMed]
  • Pou S, Pou WS, Bredt DS, Snyder SH, Rosen GM. Generation of superoxide by purified brain nitric oxide synthase. J Biol Chem. 1992 Dec 5;267(34):24173–24176. [PubMed]
  • Xia Y, Dawson VL, Dawson TM, Snyder SH, Zweier JL. Nitric oxide synthase generates superoxide and nitric oxide in arginine-depleted cells leading to peroxynitrite-mediated cellular injury. Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6770–6774. [PMC free article] [PubMed]
  • Mãrtensson J, Meister A, Mrtensson J. Glutathione deficiency decreases tissue ascorbate levels in newborn rats: ascorbate spares glutathione and protects. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4656–4660. [PMC free article] [PubMed]
  • Wink DA, Nims RW, Darbyshire JF, Christodoulou D, Hanbauer I, Cox GW, Laval F, Laval J, Cook JA, Krishna MC, et al. Reaction kinetics for nitrosation of cysteine and glutathione in aerobic nitric oxide solutions at neutral pH. Insights into the fate and physiological effects of intermediates generated in the NO/O2 reaction. Chem Res Toxicol. 1994 Jul-Aug;7(4):519–525. [PubMed]
  • Moro MA, Darley-Usmar VM, Goodwin DA, Read NG, Zamora-Pino R, Feelisch M, Radomski MW, Moncada S. Paradoxical fate and biological action of peroxynitrite on human platelets. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6702–6706. [PMC free article] [PubMed]
  • Stamler JS, Singel DJ, Loscalzo J. Biochemistry of nitric oxide and its redox-activated forms. Science. 1992 Dec 18;258(5090):1898–1902. [PubMed]
  • Stamler J, Mendelsohn ME, Amarante P, Smick D, Andon N, Davies PF, Cooke JP, Loscalzo J. N-acetylcysteine potentiates platelet inhibition by endothelium-derived relaxing factor. Circ Res. 1989 Sep;65(3):789–795. [PubMed]
  • Cooke JP, Stamler J, Andon N, Davies PF, McKinley G, Loscalzo J. Flow stimulates endothelial cells to release a nitrovasodilator that is potentiated by reduced thiol. Am J Physiol. 1990 Sep;259(3 Pt 2):H804–H812. [PubMed]
  • Creager MA, Roddy MA, Boles K, Stamler JS. N-acetylcysteine does not influence the activity of endothelium-derived relaxing factor in vivo. Hypertension. 1997 Feb;29(2):668–672. [PubMed]
  • Wong BK, Chan HC, Corcoran GB. Selective effects of N-acetylcysteine stereoisomers on hepatic glutathione and plasma sulfate in mice. Toxicol Appl Pharmacol. 1986 Dec;86(3):421–429. [PubMed]
  • Celermajer DS, Sorensen KE, Gooch VM, Spiegelhalter DJ, Miller OI, Sullivan ID, Lloyd JK, Deanfield JE. Non-invasive detection of endothelial dysfunction in children and adults at risk of atherosclerosis. Lancet. 1992 Nov 7;340(8828):1111–1115. [PubMed]
  • Sorensen KE, Celermajer DS, Georgakopoulos D, Hatcher G, Betteridge DJ, Deanfield JE. Impairment of endothelium-dependent dilation is an early event in children with familial hypercholesterolemia and is related to the lipoprotein(a) level. J Clin Invest. 1994 Jan;93(1):50–55. [PMC free article] [PubMed]
  • Celermajer DS, Sorensen KE, Bull C, Robinson J, Deanfield JE. Endothelium-dependent dilation in the systemic arteries of asymptomatic subjects relates to coronary risk factors and their interaction. J Am Coll Cardiol. 1994 Nov 15;24(6):1468–1474. [PubMed]
  • Treasure CB, Manoukian SV, Klein JL, Vita JA, Nabel EG, Renwick GH, Selwyn AP, Alexander RW, Ganz P. Epicardial coronary artery responses to acetylcholine are impaired in hypertensive patients. Circ Res. 1992 Oct;71(4):776–781. [PubMed]
  • Nitenberg A, Valensi P, Sachs R, Dali M, Aptecar E, Attali JR. Impairment of coronary vascular reserve and ACh-induced coronary vasodilation in diabetic patients with angiographically normal coronary arteries and normal left ventricular systolic function. Diabetes. 1993 Jul;42(7):1017–1025. [PubMed]
  • Nitenberg A, Antony I, Foult JM. Acetylcholine-induced coronary vasoconstriction in young, heavy smokers with normal coronary arteriographic findings. Am J Med. 1993 Jul;95(1):71–77. [PubMed]
  • Anderson TJ, Uehata A, Gerhard MD, Meredith IT, Knab S, Delagrange D, Lieberman EH, Ganz P, Creager MA, Yeung AC, et al. Close relation of endothelial function in the human coronary and peripheral circulations. J Am Coll Cardiol. 1995 Nov 1;26(5):1235–1241. [PubMed]

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