PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
 
J Clin Invest. 1986 April; 77(4): 1370–1376.
PMCID: PMC424498

Endothelial cell injury due to copper-catalyzed hydrogen peroxide generation from homocysteine.

Abstract

We have examined whether the toxic effects of homocysteine on cultured endothelial cells could result from the formation and action of hydrogen peroxide. In initial experiments with a cell-free system, micromolar amounts of copper were found to catalyze an oxygen-dependent oxidation of homocysteine. The molar ratio of homocysteine oxidized to oxygen consumed was approximately 4.0, which suggests that oxygen was reduced to water. The addition of catalase, however, decreased oxygen consumption by nearly one-half, which suggests that H2O2 was formed during the reaction. Confirming this hypothesis, H2O2 formation was detected using the horseradish peroxidase-dependent oxidation of fluorescent scopoletin. Ceruloplasmin was also found to catalyze oxidation of homocysteine and generation of H2O2 in molar amounts equivalent to copper sulfate. Finally, homocysteine oxidation was catalyzed by normal human serum in a concentration-dependent manner. Using cultured human and bovine endothelial cells, we found that homocysteine plus copper could lyse the cells in a dose-dependent manner, an effect that was completely prevented by catalase. Homocystine plus copper was not toxic to the cells. Specific injury to endothelial cells was seen only after 4 h of incubation with homocysteine plus copper. Confirming the biochemical studies, ceruloplasmin was also found to be equivalent to Cu++ in its ability to cause injury to endothelial cells in the presence of homocysteine. Since elevated levels of homocysteine have been implicated in premature development of atherosclerosis, these findings may be relevant to the mechanism of some types of chronic vascular injury.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.5M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • McCully KS. Vascular pathology of homocysteinemia: implications for the pathogenesis of arteriosclerosis. Am J Pathol. 1969 Jul;56(1):111–128. [PubMed]
  • Harker LA, Slichter SJ, Scott CR, Ross R. Homocystinemia. Vascular injury and arterial thrombosis. N Engl J Med. 1974 Sep 12;291(11):537–543. [PubMed]
  • McCully KS, Ragsdale BD. Production of arteriosclerosis by homocysteinemia. Am J Pathol. 1970 Oct;61(1):1–11. [PubMed]
  • Harker LA, Ross R, Slichter SJ, Scott CR. Homocystine-induced arteriosclerosis. The role of endothelial cell injury and platelet response in its genesis. J Clin Invest. 1976 Sep;58(3):731–741. [PMC free article] [PubMed]
  • Harker LA, Harlan JM, Ross R. Effect of sulfinpyrazone on homocysteine-induced endothelial injury and arteriosclerosis in baboons. Circ Res. 1983 Dec;53(6):731–739. [PubMed]
  • de Groot PG, Willems C, Boers GH, Gonsalves MD, van Aken WG, van Mourik JA. Endothelial cell dysfunction in homocystinuria. Eur J Clin Invest. 1983 Oct;13(5):405–410. [PubMed]
  • Wall RT, Harlan JM, Harker LA, Striker GE. Homocysteine-induced endothelial cell injury in vitro: a model for the study of vascular injury. Thromb Res. 1980 Apr 1;18(1-2):113–121. [PubMed]
  • Cavallini D, De Marco C, Duprè S, Rotilio G. The copper catalyzed oxidation of cysteine to cystine. Arch Biochem Biophys. 1969 Mar;130(1):354–361. [PubMed]
  • Sedlak J, Lindsay RH. Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem. 1968 Oct 24;25(1):192–205. [PubMed]
  • Starkebaum G, Root RK. D-Penicillamine: analysis of the mechanism of copper-catalyzed hydrogen peroxide generation. J Immunol. 1985 May;134(5):3371–3378. [PubMed]
  • Root RK, Metcalf J, Oshino N, Chance B. H2O2 release from human granulocytes during phagocytosis. I. Documentation, quantitation, and some regulating factors. J Clin Invest. 1975 May;55(5):945–955. [PMC free article] [PubMed]
  • Harlan JM, Killen PD, Harker LA, Striker GE, Wright DG. Neutrophil-mediated endothelial injury in vitro mechanisms of cell detachment. J Clin Invest. 1981 Dec;68(6):1394–1403. [PMC free article] [PubMed]
  • Ross R, Glomset JA. Atherosclerosis and the arterial smooth muscle cell: Proliferation of smooth muscle is a key event in the genesis of the lesions of atherosclerosis. Science. 1973 Jun 29;180(4093):1332–1339. [PubMed]
  • Harlan JM, Levine JD, Callahan KS, Schwartz BR, Harker LA. Glutathione redox cycle protects cultured endothelial cells against lysis by extracellularly generated hydrogen peroxide. J Clin Invest. 1984 Mar;73(3):706–713. [PMC free article] [PubMed]
  • Misra HP. Generation of superoxide free radical during the autoxidation of thiols. J Biol Chem. 1974 Apr 10;249(7):2151–2155. [PubMed]
  • Saez G, Thornalley PJ, Hill HA, Hems R, Bannister JV. The production of free radicals during the autoxidation of cysteine and their effect on isolated rat hepatocytes. Biochim Biophys Acta. 1982 Oct 28;719(1):24–31. [PubMed]
  • Chidambaram MV, Zgirski A, Frieden E. The reaction of cysteine with ceruloplasmin copper. J Inorg Biochem. 1984 Jul;21(3):227–239. [PubMed]
  • Nyberg GK, Granberg GP, Carlsson J. Bovine superoxide dismutase and copper ions potentiate the bactericidal effect of autoxidizing cysteine. Appl Environ Microbiol. 1979 Jul;38(1):29–34. [PMC free article] [PubMed]
  • Glatt H, Protić-SabljiC M, Oesch F. Mutagenicity of glutathione and cysteine in the Ames test. Science. 1983 May 27;220(4600):961–963. [PubMed]
  • O'Donnell-Tormey J, DeBoer CJ, Nathan CF. Resistance of human tumor cells in vitro to oxidative cytolysis. J Clin Invest. 1985 Jul;76(1):80–86. [PMC free article] [PubMed]
  • Ross R, Glomset JA. The pathogenesis of atherosclerosis (second of two parts). N Engl J Med. 1976 Aug 19;295(8):420–425. [PubMed]
  • Dudman NP, Wilcken DE. Increased plasma copper in patients with homocystinuria due to cystathionine beta-synthase deficiency. Clin Chim Acta. 1983 Jan 7;127(1):105–113. [PubMed]
  • Brattstrom LE, Hardebo JE, Hultberg BL. Moderate homocysteinemia--a possible risk factor for arteriosclerotic cerebrovascular disease. Stroke. 1984 Nov-Dec;15(6):1012–1016. [PubMed]
  • Boers GH, Smals AG, Trijbels FJ, Fowler B, Bakkeren JA, Schoonderwaldt HC, Kleijer WJ, Kloppenborg PW. Heterozygosity for homocystinuria in premature peripheral and cerebral occlusive arterial disease. N Engl J Med. 1985 Sep 19;313(12):709–715. [PubMed]
  • Wilcken DE, Gupta VJ. Cysteine-homocysteine mixed disulphide: differing plasma concentrations in normal men and women. Clin Sci (Lond) 1979 Aug;57(2):211–215. [PubMed]
  • Gupta VJ, Wilcken DE. The detection of cysteine-homocysteine mixed disulphide in plasma of normal fasting man. Eur J Clin Invest. 1978 Aug;8(4):205–207. [PubMed]
  • Boers GH, Smals AG, Trijbels FJ, Leermakers AI, Kloppenborg PW. Unique efficiency of methionine metabolism in premenopausal women may protect against vascular disease in the reproductive years. J Clin Invest. 1983 Dec;72(6):1971–1976. [PMC free article] [PubMed]
  • Wilcken DE, Gupta VJ. Sulphr containing amino acids in chronic renal failure with particular reference to homocystine and cysteine-homocysteine mixed disulphide. Eur J Clin Invest. 1979 Aug;9(4):301–307. [PubMed]
  • Wilcken DE, Gupta VJ, Reddy SG. Accumulation of sulphur-containing amino acids including cysteine-homocysteine in patients on maintenance haemodialysis. Clin Sci (Lond) 1980 May;58(5):427–430. [PubMed]
  • Wilcken DE, Gupta VJ, Betts AK. Homocysteine in the plasma of renal transplant recipients: effects of cofactors for methionine metabolism. Clin Sci (Lond) 1981 Dec;61(6):743–749. [PubMed]
  • Warso MA, Lands WE. Presence of lipid hydroperoxide in human plasma. J Clin Invest. 1985 Feb;75(2):667–671. [PMC free article] [PubMed]
  • Taylor L, Menconi MJ, Polgar P. The participation of hydroperoxides and oxygen radicals in the control of prostaglandin synthesis. J Biol Chem. 1983 Jun 10;258(11):6855–6857. [PubMed]
  • Whorton AR, Montgomery ME, Kent RS. Effect of hydrogen peroxide on prostaglandin production and cellular integrity in cultured porcine aortic endothelial cells. J Clin Invest. 1985 Jul;76(1):295–302. [PMC free article] [PubMed]
  • Heinecke JW, Rosen H, Chait A. Iron and copper promote modification of low density lipoprotein by human arterial smooth muscle cells in culture. J Clin Invest. 1984 Nov;74(5):1890–1894. [PMC free article] [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation