Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1984 Feb 1;159(2):592–603. doi: 10.1084/jem.159.2.592

Differential effects of hydrogen peroxide on indices of endothelial cell function

PMCID: PMC2187215  PMID: 6363599

Abstract

The responses of pig aortic endothelial cells to sublethal doses of potentially toxic stimuli were investigated by monitoring K+ efflux, prostaglandin production, and the release of cytoplasmic purines. Xanthine plus xanthine oxidase reversibly stimulated these three parameters of endothelial cell function at doses that were not cytotoxic, as measured by chromium release, adenine uptake, and vital dye exclusion. The effects of xanthine plus xanthine oxidase were inhibited by catalase but not by superoxide dismutase, suggesting that H2O2 was responsible. Reagent H2O2 also reversibly stimulated K+ efflux, prostaglandin production, and the release of purines. The threshold concentration of H2O2 for these effects was approximately 10 microM, which was at least 30-fold lower than that which caused cytotoxicity. In addition to the direct effect of H2O2 in stimulating prostaglandin production (PGI2 and PGE2), prior exposure of endothelial cells to lower doses of H2O2 (less than 0.1 microM) at high oxygen tension inhibited the subsequent stimulation of prostaglandin production by ATP, A23187, and H2O2 itself. We conclude that H2O2 has substantial effects on endothelial physiology at doses up to 3,000-fold lower than those which induce cytotoxicity.

Full Text

The Full Text of this article is available as a PDF (943.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ager A., Gordon J. L., Moncada S., Pearson J. D., Salmon J. A., Trevethick M. A. Effects of isolation and culture on prostaglandin synthesis by porcine aortic endothelial and smooth muscle cells. J Cell Physiol. 1982 Jan;110(1):9–16. doi: 10.1002/jcp.1041100103. [DOI] [PubMed] [Google Scholar]
  2. Arrick B. A., Nathan C. F., Griffith O. W., Cohn Z. A. Glutathione depletion sensitizes tumor cells to oxidative cytolysis. J Biol Chem. 1982 Feb 10;257(3):1231–1237. [PubMed] [Google Scholar]
  3. COHEN G., HOCHSTEIN P. GLUTATHIONE PEROXIDASE: THE PRIMARY AGENT FOR THE ELIMINATION OF HYDROGEN PEROXIDE IN ERYTHROCYTES. Biochemistry. 1963 Nov-Dec;2:1420–1428. doi: 10.1021/bi00906a038. [DOI] [PubMed] [Google Scholar]
  4. Egan R. W., Gale P. H., Kuehl F. A., Jr Reduction of hydroperoxides in the prostaglandin biosynthetic pathway by a microsomal peroxidase. J Biol Chem. 1979 May 10;254(9):3295–3302. [PubMed] [Google Scholar]
  5. Egan R. W., Paxton J., Kuehl F. A., Jr Mechanism for irreversible self-deactivation of prostaglandin synthetase. J Biol Chem. 1976 Dec 10;251(23):7329–7335. [PubMed] [Google Scholar]
  6. Gordon J. L., Martin W. Endothelium-dependent relaxation of the pig aorta: relationship to stimulation of 86Rb efflux from isolated endothelial cells. Br J Pharmacol. 1983 Jun;79(2):531–541. doi: 10.1111/j.1476-5381.1983.tb11028.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gryglewski R. J., Bunting S., Moncada S., Flower R. J., Vane J. R. Arterial walls are protected against deposition of platelet thrombi by a substance (prostaglandin X) which they make from prostaglandin endoperoxides. Prostaglandins. 1976 Nov;12(5):685–713. doi: 10.1016/0090-6980(76)90047-2. [DOI] [PubMed] [Google Scholar]
  8. Ham E. A., Egan R. W., Soderman D. D., Gale P. H., Kuehl F. A., Jr Peroxidase-dependent deactivation of prostacyclin synthetase. J Biol Chem. 1979 Apr 10;254(7):2191–2194. [PubMed] [Google Scholar]
  9. Hemler M. E., Lands W. E. Evidence for a peroxide-initiated free radical mechanism of prostaglandin biosynthesis. J Biol Chem. 1980 Jul 10;255(13):6253–6261. [PubMed] [Google Scholar]
  10. Hong S. L., Carty T., Deykin D. Tranylcypromine and 15-hydroperoxyarachidonate affect arachidonic acid release in addition to inhibition of prostacyclin synthesis in calf aortic endothelial cells. J Biol Chem. 1980 Oct 25;255(20):9538–9540. [PubMed] [Google Scholar]
  11. Hong S. L., Deykin D. Activation of phospholipases A2 and C in pig aortic endothelial cells synthesizing prostacyclin. J Biol Chem. 1982 Jun 25;257(12):7151–7154. [PubMed] [Google Scholar]
  12. Hong S. L. Effect of bradykinin and thrombin on prostacyclin synthesis in endothelial cells from calf and pig aorta and human umbilical cord vein. Thromb Res. 1980 Jun 15;18(6):787–795. doi: 10.1016/0049-3848(80)90201-7. [DOI] [PubMed] [Google Scholar]
  13. Lollar P., Owen W. G. Active-site-dependent, thrombin-induced release of adenine nucleotides from cultured human endothelial cells. Ann N Y Acad Sci. 1981;370:51–56. doi: 10.1111/j.1749-6632.1981.tb29720.x. [DOI] [PubMed] [Google Scholar]
  14. Martin W., Gordon J. L. Differential calcium dependence of contractile responses and 86Rb efflux from the rabbit aorta induced by vasoactive stimuli. J Cell Physiol. 1983 Apr;115(1):46–52. doi: 10.1002/jcp.1041150108. [DOI] [PubMed] [Google Scholar]
  15. Nathan C. F., Brukner L. H., Silverstein S. C., Cohn Z. A. Extracellular cytolysis by activated macrophages and granulocytes. I. Pharmacologic triggering of effector cells and the release of hydrogen peroxide. J Exp Med. 1979 Jan 1;149(1):84–99. doi: 10.1084/jem.149.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pearson J. D., Carleton J. S., Gordon J. L. Metabolism of adenine nucleotides by ectoenzymes of vascular endothelial and smooth-muscle cells in culture. Biochem J. 1980 Aug 15;190(2):421–429. doi: 10.1042/bj1900421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pearson J. D., Carleton J. S., Hutchings A., Gordon J. L. Uptake and metabolism of adenosine by pig aortic endothelial and smooth-muscle cells in culture. Biochem J. 1978 Feb 15;170(2):265–271. doi: 10.1042/bj1700265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pearson J. D., Carleton J. S., Hutchings A. Prostacyclin release stimulated by thrombin or bradykinin in porcine endothelial cells cultured from aorta and umbilical vein. Thromb Res. 1983 Jan 15;29(2):115–124. doi: 10.1016/0049-3848(83)90133-0. [DOI] [PubMed] [Google Scholar]
  19. Pearson J. D., Gordon J. L. Vascular endothelial and smooth muscle cells in culture selectively release adenine nucleotides. Nature. 1979 Oct 4;281(5730):384–386. doi: 10.1038/281384a0. [DOI] [PubMed] [Google Scholar]
  20. Polgar P., Taylor L. Stimulation of prostaglandin synthesis by ascorbic acid via hydrogen peroxide formation. Prostaglandins. 1980 May;19(5):693–700. doi: 10.1016/0090-6980(80)90168-9. [DOI] [PubMed] [Google Scholar]
  21. Rosen H., Klebanoff S. J. Bactericidal activity of a superoxide anion-generating system. A model for the polymorphonuclear leukocyte. J Exp Med. 1979 Jan 1;149(1):27–39. doi: 10.1084/jem.149.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sacks T., Moldow C. F., Craddock P. R., Bowers T. K., Jacob H. S. Oxygen radicals mediate endothelial cell damage by complement-stimulated granulocytes. An in vitro model of immune vascular damage. J Clin Invest. 1978 May;61(5):1161–1167. doi: 10.1172/JCI109031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Smith W. L., Lands W. E. Oxygenation of polyunsaturated fatty acids during prostaglandin biosynthesis by sheep vesicular gland. Biochemistry. 1972 Aug 15;11(17):3276–3285. doi: 10.1021/bi00767a024. [DOI] [PubMed] [Google Scholar]
  24. Taylor L., Menconi M. J., 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] [Google Scholar]
  25. WROBLEWSKI F., LADUE J. S. Lactic dehydrogenase activity in blood. Proc Soc Exp Biol Med. 1955 Oct;90(1):210–213. doi: 10.3181/00379727-90-21985. [DOI] [PubMed] [Google Scholar]
  26. Waud W. R., Brady F. O., Wiley R. D., Rajagopalan K. V. A new purification procedure for bovine milk xanthine oxidase: effect of proteolysis on the subunit structure. Arch Biochem Biophys. 1975 Aug;169(2):695–701. doi: 10.1016/0003-9861(75)90214-3. [DOI] [PubMed] [Google Scholar]
  27. Wedmore C. V., Williams T. J. Control of vascular permeability by polymorphonuclear leukocytes in inflammation. Nature. 1981 Feb 19;289(5799):646–650. doi: 10.1038/289646a0. [DOI] [PubMed] [Google Scholar]
  28. Weening R. S., Wever R., Roos D. Quantitative aspects of the production of superoxide radicals by phagocytizing human granulocytes. J Lab Clin Med. 1975 Feb;85(2):245–252. [PubMed] [Google Scholar]
  29. Weiss S. J., Young J., LoBuglio A. F., Slivka A., Nimeh N. F. Role of hydrogen peroxide in neutrophil-mediated destruction of cultured endothelial cells. J Clin Invest. 1981 Sep;68(3):714–721. doi: 10.1172/JCI110307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weksler B. B., Ley C. W., Jaffe E. A. Stimulation of endothelial cell prostacyclin production by thrombin, trypsin, and the ionophore A 23187. J Clin Invest. 1978 Nov;62(5):923–930. doi: 10.1172/JCI109220. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES