Skip to main content
Log in

Effects of peroxide on endothelial nitric oxide synthase in coronary arteries

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Reactive oxygen species (ROS) are produced in ischemia and reperfusion. Since endothelial nitric oxide synthase (eNOS) is key to the endothelium- dependent vasodilation, we examined the effects of peroxide on this enzyme. We treated cells cultured from pig coronary artery endothelium with different concentrations of hydrogen peroxide, washed them, solubilized them and measured NOS activity by arginine to citrulline conversion. Hydrogen peroxide inhibited the eNOS activity with an IC50 value of 0.85 ± 0.39 mM. In another experiment, we perfused arteries with solutions containing 0 or 1 mM hydrogen peroxide, washed them, removed the endothelium using a cotton swab, centrifuged and solubilized the endothelium and monitored its NOS activity. Hydrogen peroxide (1 mM) did not affect the NOS activity significantly (p > 0.05) in this assay. We conclude that the inactivation of eNOS by hydrogen peroxide does not play a major role in the ischemia- reperfusion damage because the peroxide concentrations attained during ischemia-perfusion are much lower than those affecting the eNOS activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Pantely GA, Bristow JD, Swenson LJ, Ladley HD, Johnson WB, Anselone CG. Incomplete coronary vasodilation during myocardial ischemia in swine. Am J Physiol 249: H638–H647, 1985

    PubMed  Google Scholar 

  2. Sobey CG, Woodman OL: Myocardial ischaemia-What happens to the coronary arteries? Trends Pharmacol Sci 14: 448–453, 1993

    Article  PubMed  Google Scholar 

  3. Detry JM: The pathophysiology of myocardial ischaemia. Eur Heart J 17 Suppl G: 48–52, 1996

    Google Scholar 

  4. Singal PK, Dhalla AK, Hill M, Thomas TP: Endogenous antioxidant changes in the myocardium in response to acute and chronic stress conditions. Mol Cell Biochem 129: 179–186, 1993

    PubMed  Google Scholar 

  5. Downey JM: Free radicals and their involvement during long-term myocardial ischemia and reperfusion. [Review]. Ann Rev Physiol 52: 487–504, 1990

    Article  Google Scholar 

  6. Kim MS, Akera T: O2 free radicals: cause of ischemia-reperfusion injury to cardiac Na+-K+-ATPase. Am J Physiol 252: H252–H257, 1987

    PubMed  Google Scholar 

  7. Globus MYT, Busto R, Lin BW, Schnippering H, Ginsberg MD: Detection of free radical activity during transient global ischemia and recirculation: Effects of intraischemic brain temperature modulation. J Neurochem 65: 1250–1256, 1995

    PubMed  Google Scholar 

  8. Suzuki YJ, Ford GD: Inhibition of Ca(2+)-ATPase of vascular smooth muscle sarcoplasmic reticulum by reactive oxygen intermediates. Am J Physiol 261: H568–H574, 1991

    PubMed  Google Scholar 

  9. Luscher TF, Tanner FC, Tschudi MR, Noll G: Endothelial dysfunction in coronary artery disease. [Review]. Ann Rev Med 44: 395–418, 1993

    Article  PubMed  Google Scholar 

  10. Grover AK, Samson SE, Fomin VP: Peroxide inactivates calcium pumps in pig coronary artery. Am J Physiol 263: H537–H543, 1992

    PubMed  Google Scholar 

  11. Roveri A, Coassin M, Maiorino M, Zamburlini A, van Amsterdam FT, Ursini F: Effect of hydrogen peroxide on calcium homeostasis in smooth muscle cells. Arch Biochem Biophys 297: 265–70, 1992

    PubMed  Google Scholar 

  12. Todoki K, Okabe E, Kiyose T, Sekishita T, Ito H: Oxygen free radicalmediated selective endothelial dysfunction in isolated coronary artery. Am J Physiol 262: H806–H812, 1992

    PubMed  Google Scholar 

  13. Gross GJ, O'Rourke ST, Pelc LR, Warltier DC: Myocardial and endothelial dysfunction after multiple, brief coronary occlusions: role of oxygen radicals. Am J Physiol 263: H1703–H1709, 1992

    PubMed  Google Scholar 

  14. Bradley JR, Johnson DR, Pober JS: Endothelial activation by hydrogen peroxide. Selective increases of intercellular adhesion molecule-1 and major histocompatibility complex class I. Am J Path 142: 1598–1609, 1993

    PubMed  Google Scholar 

  15. Kukreja RC, Okabe E, Schrier GM, Hess ML: Oxygen radicalmediated lipid peroxidation and inhibition of Ca2+-ATPase activity of cardiac sarcoplasmic reticulum. Arch Biochem Biophys 261: 447–457, 1988

    PubMed  Google Scholar 

  16. Dhalla NS, Dixon IM, Rupp H, Barwinsky J: Experimental congestive heart failure due to myocardial infarction: sarcolemmal receptors and cation transporters. Basic Res Cardiol 86 Suppl 3: 13–23, 1991

    Google Scholar 

  17. Grover AK, Samson SE, Fomin VP, Werstiuk ES: Effects of peroxide and superoxide on coronary artery: Ang II response and sarcoplasmic reticulum Ca2+-pump. Am J Physiol 269: C546–C553, 1995

    PubMed  Google Scholar 

  18. Elmoselhi A, Butcher A, Samson SE, Grover AK: Free radicals uncouple the sodium pump in pig coronary artery. Am J Physiol 266: C720–C728, 1994

    PubMed  Google Scholar 

  19. Vlessis AA, Muller P, Bartos D, Trunkey D: Mechanism of peroxide-induced cellular injury in cultured adult cardiac myocytes. FASEB J 5: 2600–2605, 1991

    PubMed  Google Scholar 

  20. Duprat F, Guillemare E, Romey G, Fink M, Lesage F, Lazdunski M, Honore E: Susceptibility of cloned K+ channels to reactive oxygen species. Proc Natl Acad Sci USA 92: 11796–11800, 1995

    PubMed  Google Scholar 

  21. Kiechle FL, Malinski T: Nitric oxide. Biochemistry, pathophysiology, and detection. Am J Clin Pathol 100: 567–575, 1993

    PubMed  Google Scholar 

  22. Bredt DS, Hwang PM, Glatt CE, Lowenstein C, Reed RR, Snyder SH: Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase. Nature 351: 714–718, 1991

    Article  PubMed  Google Scholar 

  23. Lamas S, Marsden PA, Li GK, Tempst P, Michel T: Endothelial nitric oxide synthase: molecular cloning and characterization of a distinct constitutive enzyme isoform. Proc Nat Acad Sci USA 89: 6348–6352, 1992

    PubMed  Google Scholar 

  24. Grover AK, Samson SE: Sarco/endoplasmic reticulum Ca2+ pump is more resistant to peroxide in endothelium than in smooth muscle: implications to coronary artery function. Am J Physiol (accepted) 1997

  25. Grover AK, Samson SE, Lee RM: Subcellular fractionation of pig coronary artery smooth muscle. Biochim Biophys Acta 818: 191–199, 1985

    PubMed  Google Scholar 

  26. Elmoselhi A, Samson SE, Grover AK: Properties of the sarcoplasmic reticulum Ca2+-pump in coronary artery skinned smooth muscle. Mol Cell Biochem 151: 149–155, 1995

    PubMed  Google Scholar 

  27. Bredt DS, Snyder SH: Isolation of nitric oxide synthetase, a calmodulinrequiring enzyme. Proc Natl Acad Sci USA 87: 682–685, 1990

    PubMed  Google Scholar 

  28. Kostka P, Jang E, Watson EG, Stewart JL, Daniel EE: Nitric oxide synthase in the autonomic nervous system of canine ileum. J Pharmacol Exp Ther 264: 234–239, 1993

    PubMed  Google Scholar 

  29. Darley-Usmar V, Wiseman H, Halliwell B: Nitric oxide and oxygen radicals: a question of balance. FEBS Lett 369: 131–135, 1995

    Article  PubMed  Google Scholar 

  30. Tsao PS, Lefer AM: Time course and mechanism of endothelial dysfunction in isolated ischemic-and hypoxic-perfused rat hearts. Am J Physiol 259: H1660–H1666, 1990

    PubMed  Google Scholar 

  31. Shattock MJ, Manning AS, Hearse DJ: Effects of hydrogen peroxide on cardiac function and post-ischaemic functional recovery in the isolated ‘working’ rat heart. Pharmacology 24: 118–122, 1982

    PubMed  Google Scholar 

  32. Mukhopadhyay A, Steinberg N, Das DK: Effects of free radicals and oxidants on myocardial cellular injury. Clin Physiol Biochem 7: 278–285, 1989

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shah, K., Samson, S. & Grover, A. Effects of peroxide on endothelial nitric oxide synthase in coronary arteries. Mol Cell Biochem 183, 147–152 (1998). https://doi.org/10.1023/A:1006828205261

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006828205261

Navigation