Endogenous Peroxynitrite Is Involved in the Inhibition of Mitochondrial Respiration in Immuno-Stimulated J774.2 Macrophages

Csaba Szabo, A. L. Salzman

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111 Citations (Scopus)

Abstract

The free radicals nitric oxide (NO) and surperoxide (O2 -) are known to react to form peroxynitrite (ONOO-), a potentially more injurious species. Here we compared the inhibitory effects of ONOO- and NO on mitochondrial respiration in J774.2 macrophages. In addition, using uric acid, a potent scavenger of ONOO-, we investigated the potential involvement of endogenous ONOO- in the inhibitory effects of bacterial lipopolysaccharide (LPS) and γ-interferon (IFN) on mitochondrial respiration. The NO donors S-nitroso-N-acetyl-DL-penicillamine (SNAP, 1 mM) or diethylamine NONOate (DN, 1 mM) inhibited cellular respiration by approximately 30% over 24h. Equimolar amounts of ONOO- caused a more pronounced inhibition of cell respiration. There was a synergistic effect between the O2 - generator pyrogallol (10 μM-1 mM) and the NO donor SNAP (1 mM) in inhibiting mitochondrial respiration. The ONOO- scavenger uric acid (UA, 1 mM) did not prevent the decrease in viability in response to SNAP, DN or pyrogallol, but significantly prevented the decrease in cell viability in response to ONOO-, to the combination of SNAP and pyrogallol, and to SIN-1, a compound that simultaneously generates NO and O2 -. The decrease in mitochondrial respiration in response to LPS and IFN was also inhibited by UA as well as by NG-methyl-arginine, an inhibitor of NOS. Thus, ONOO- is a more potent suppressant of mitochondrial respiration than NO and endogenous formation of ONOO- appears to be involved in the cytotoxicity associated with immune stimulation.

Original languageEnglish (US)
Pages (from-to)739-743
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume209
Issue number2
DOIs
StatePublished - Apr 17 1995
Externally publishedYes

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Peroxynitrous Acid
Macrophages
Pyrogallol
Nitric Oxide
Respiration
Nitric Oxide Donors
Cell Respiration
Uric Acid
Interferons
Lipopolysaccharides
Cells
Penicillamine
Cytotoxicity
Free Radicals
Arginine
Cell Survival

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology
  • Biophysics
  • Biochemistry

Cite this

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title = "Endogenous Peroxynitrite Is Involved in the Inhibition of Mitochondrial Respiration in Immuno-Stimulated J774.2 Macrophages",
abstract = "The free radicals nitric oxide (NO) and surperoxide (O2 -) are known to react to form peroxynitrite (ONOO-), a potentially more injurious species. Here we compared the inhibitory effects of ONOO- and NO on mitochondrial respiration in J774.2 macrophages. In addition, using uric acid, a potent scavenger of ONOO-, we investigated the potential involvement of endogenous ONOO- in the inhibitory effects of bacterial lipopolysaccharide (LPS) and γ-interferon (IFN) on mitochondrial respiration. The NO donors S-nitroso-N-acetyl-DL-penicillamine (SNAP, 1 mM) or diethylamine NONOate (DN, 1 mM) inhibited cellular respiration by approximately 30{\%} over 24h. Equimolar amounts of ONOO- caused a more pronounced inhibition of cell respiration. There was a synergistic effect between the O2 - generator pyrogallol (10 μM-1 mM) and the NO donor SNAP (1 mM) in inhibiting mitochondrial respiration. The ONOO- scavenger uric acid (UA, 1 mM) did not prevent the decrease in viability in response to SNAP, DN or pyrogallol, but significantly prevented the decrease in cell viability in response to ONOO-, to the combination of SNAP and pyrogallol, and to SIN-1, a compound that simultaneously generates NO and O2 -. The decrease in mitochondrial respiration in response to LPS and IFN was also inhibited by UA as well as by NG-methyl-arginine, an inhibitor of NOS. Thus, ONOO- is a more potent suppressant of mitochondrial respiration than NO and endogenous formation of ONOO- appears to be involved in the cytotoxicity associated with immune stimulation.",
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AU - Salzman, A. L.

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N2 - The free radicals nitric oxide (NO) and surperoxide (O2 -) are known to react to form peroxynitrite (ONOO-), a potentially more injurious species. Here we compared the inhibitory effects of ONOO- and NO on mitochondrial respiration in J774.2 macrophages. In addition, using uric acid, a potent scavenger of ONOO-, we investigated the potential involvement of endogenous ONOO- in the inhibitory effects of bacterial lipopolysaccharide (LPS) and γ-interferon (IFN) on mitochondrial respiration. The NO donors S-nitroso-N-acetyl-DL-penicillamine (SNAP, 1 mM) or diethylamine NONOate (DN, 1 mM) inhibited cellular respiration by approximately 30% over 24h. Equimolar amounts of ONOO- caused a more pronounced inhibition of cell respiration. There was a synergistic effect between the O2 - generator pyrogallol (10 μM-1 mM) and the NO donor SNAP (1 mM) in inhibiting mitochondrial respiration. The ONOO- scavenger uric acid (UA, 1 mM) did not prevent the decrease in viability in response to SNAP, DN or pyrogallol, but significantly prevented the decrease in cell viability in response to ONOO-, to the combination of SNAP and pyrogallol, and to SIN-1, a compound that simultaneously generates NO and O2 -. The decrease in mitochondrial respiration in response to LPS and IFN was also inhibited by UA as well as by NG-methyl-arginine, an inhibitor of NOS. Thus, ONOO- is a more potent suppressant of mitochondrial respiration than NO and endogenous formation of ONOO- appears to be involved in the cytotoxicity associated with immune stimulation.

AB - The free radicals nitric oxide (NO) and surperoxide (O2 -) are known to react to form peroxynitrite (ONOO-), a potentially more injurious species. Here we compared the inhibitory effects of ONOO- and NO on mitochondrial respiration in J774.2 macrophages. In addition, using uric acid, a potent scavenger of ONOO-, we investigated the potential involvement of endogenous ONOO- in the inhibitory effects of bacterial lipopolysaccharide (LPS) and γ-interferon (IFN) on mitochondrial respiration. The NO donors S-nitroso-N-acetyl-DL-penicillamine (SNAP, 1 mM) or diethylamine NONOate (DN, 1 mM) inhibited cellular respiration by approximately 30% over 24h. Equimolar amounts of ONOO- caused a more pronounced inhibition of cell respiration. There was a synergistic effect between the O2 - generator pyrogallol (10 μM-1 mM) and the NO donor SNAP (1 mM) in inhibiting mitochondrial respiration. The ONOO- scavenger uric acid (UA, 1 mM) did not prevent the decrease in viability in response to SNAP, DN or pyrogallol, but significantly prevented the decrease in cell viability in response to ONOO-, to the combination of SNAP and pyrogallol, and to SIN-1, a compound that simultaneously generates NO and O2 -. The decrease in mitochondrial respiration in response to LPS and IFN was also inhibited by UA as well as by NG-methyl-arginine, an inhibitor of NOS. Thus, ONOO- is a more potent suppressant of mitochondrial respiration than NO and endogenous formation of ONOO- appears to be involved in the cytotoxicity associated with immune stimulation.

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