Poly(ADP-ribose) synthetase activation mediates mitochondrial injury during oxidant-induced cell death

László Virág, Andrew L. Salzman, Csaba Szabo

Research output: Contribution to journalArticle

200 Citations (Scopus)

Abstract

Reactive oxidant species are important mediators of tissue injury in shock, inflammation, and reperfusion injury. The actions of a number of these oxidants (e.g., hydroxyl radical and peroxynitrite, a reactive oxidant produced by the reaction of nitric oxide and superoxide) are mediated in part by the activation of the nuclear nick sensor enzyme, poly(ADP)-ribose synthetase (PARS), with consequent cellular energy depletion. Here we investigated whether PARS activation contributes to the mitochondrial alterations in cells exposed to oxidants. Authentic peroxynitrite (20 μM), the peroxynitrite-generating compound 3-morpholinosid-nonimine, the combination of pyrogallol and S-nitroso-N-acetyl-D,L-penicillamine, as well as hydrogen peroxide induced a time-and dose-dependent decrease in mitochondrial transmembrane potential (ΔΩ(m)) in thymocytes, as determined by flow cytometry using the mitochondrial potential sensitive dyes DiOC6(3) and JC-1. A time- and dose-dependent increase in secondary reactive oxygen intermediate production and loss of cardiolipin, an indicator of mitochondrial membrane damage, were also observed, as measured by flow cytometry using the fluorescent dyes dihydroethidine and nonyl-acridine orange, respectively. Inhibition of PARS by 3-aminobenzamide or 5-iodo-6- amino-1,2-benzopyrone attenuated peroxynitrite-induced ΔΩ(m) reduction, secondary reactive oxygen intermediate generation, cardiolipin degradation, and intracellular calcium mobilization. Furthermore, thymocytes from PARS- deficient animals were protected against the peroxynitrite- and hydrogen peroxide-induced functional and ultrastructural mitochondrial alterations. In conclusion, mitochondrial perturbations during oxidant-mediated cytotoxicity are, to a significant degree, related to PARS activation rather than to direct effects of the oxidants on the mitochondria.

Original languageEnglish (US)
Pages (from-to)3753-3759
Number of pages7
JournalJournal of Immunology
Volume161
Issue number7
StatePublished - Oct 1 1998
Externally publishedYes

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Poly Adenosine Diphosphate Ribose
Ligases
Peroxynitrous Acid
Oxidants
Cell Death
Wounds and Injuries
Cardiolipins
Thymocytes
Hydrogen Peroxide
Flow Cytometry
Pyrogallol
Oxygen
Acridine Orange
Penicillamine
Mitochondrial Membranes
Reperfusion Injury
Fluorescent Dyes
Superoxides
Hydroxyl Radical
Membrane Potentials

ASJC Scopus subject areas

  • Immunology

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Poly(ADP-ribose) synthetase activation mediates mitochondrial injury during oxidant-induced cell death. / Virág, László; Salzman, Andrew L.; Szabo, Csaba.

In: Journal of Immunology, Vol. 161, No. 7, 01.10.1998, p. 3753-3759.

Research output: Contribution to journalArticle

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