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Related Experiment Videos

Peroxynitrite formation from macrophage-derived nitric oxide.

H Ischiropoulos1, L Zhu, J S Beckman

  • 1Department of Anesthesiology, University of Alabama, Birmingham 35294.

Archives of Biochemistry and Biophysics
|November 1, 1992
PubMed
Summary

Activated macrophages produce peroxynitrite, a potent oxidant implicated in tissue injury. This study quantifies peroxynitrite formation and its contribution to inflammation.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Immunology

Background:

  • Macrophages play a key role in inflammatory responses.
  • Nitric oxide (NO) and superoxide are key mediators in macrophage activation.
  • Peroxynitrite, formed from NO and superoxide, is a potent oxidant.

Purpose of the Study:

  • To quantify peroxynitrite formation by activated rat alveolar macrophages.
  • To investigate the role of superoxide dismutase (SOD) in peroxynitrite detection.
  • To assess the contribution of peroxynitrite to inflammatory processes.

Main Methods:

  • Assay of peroxynitrite formation using Cu,Zn SOD-catalyzed nitration of 4-hydroxyphenylacetate.
  • Inhibition studies using N-methyl-L-arginine, an inhibitor of nitric oxide synthesis.

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  • Measurement of superoxide and nitrite/nitrate accumulation.

    • Main Results:

    • Activated macrophages produced peroxynitrite at a rate of 9 +/- 2 pmol x 10(6) cells-1 x min-1.
    • N-methyl-L-arginine inhibited peroxynitrite formation, confirming its dependence on nitric oxide synthesis.
    • Cu-depleted Zn SOD failed to catalyze nitration, highlighting the role of Cu,Zn SOD.
    • Peroxynitrite formation rate was estimated up to 0.11 nmol x 10(6) cells-1 x min-1.
    • Nitrite and nitrate accumulation indicated significant nitric oxide conversion to peroxynitrite.

    Conclusions:

    • Activated macrophages generate significant amounts of peroxynitrite.
    • Peroxynitrite is a major product of nitric oxide metabolism in activated macrophages.
    • The formation of this strong oxidant may contribute to inflammatory cell-mediated tissue injury.