Increased spontaneous chemiluminescence from liver homogenates and isolated hepatocytes upon inhibition of o2 - and h2o2 utilization

Julio F. Turrens, Cecilia R Giulivi, Alberto Boveris

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The intracellular steady-state concentrations of hydrogen peroxide or Superoxide anion were increased by inhibiting either catalase, glutathione peroxidase, or Superoxide dismutase activities. Catalase was inhibited with aminotriazole while glutathione peroxidase activity was blocked by eliminating reduced glutathione after addition of either iodoacetamide diethylmaleate or phorone. The concentration of aminotriazole that stimulated chemiluminescence in 50% (60 mM) was very similar to the Ki for catalase activity (70 mM). Cyanide, an inhibitor of both catalase and Superoxide dismutase, stimulated chemiluminescence in 50% at a concentration (0.15 mM) which is much closer from the Ki for Superoxide dismutase (0.25 mM) than from the Ki for catalase (15 μM). The Superoxide dismutase inhibitor diethyldithiocarbamate also increased chemiluminescence six- to ten-fold. Depletion of reduced glutathione stimulated spontaneous chemiluminescence when its concentration decreased below 4.5 μmol · g liver-1. The results shown herein suggest that the changes in the intracellular steady-state concentration occurring after inhibition of any antioxidant enzyme are responsible for the increased spontaneous chemilumi-nescence. Spontaneous chemiluminescence from intact cells may be used as a noninvasive method for monitoring intracellular free radical metabolism.

Original languageEnglish (US)
Pages (from-to)135-140
Number of pages6
JournalJournal of Free Radicals in Biology and Medicine
Volume2
Issue number2
DOIs
StatePublished - 1986
Externally publishedYes

Fingerprint

Chemiluminescence
Luminescence
Liver
Catalase
Hepatocytes
Superoxide Dismutase
Amitrole
diethyl maleate
Glutathione Peroxidase
Glutathione
Iodoacetamide
Ditiocarb
Cyanides
Metabolism
Superoxides
Hydrogen Peroxide
Free Radicals
Antioxidants
Monitoring
Enzymes

Keywords

  • Aminotriazole
  • Antioxidant enzymes
  • Catalase
  • Chemiluminescence
  • Glutathione peroxidase
  • Lipid peroxidation
  • Superoxide dismutase

ASJC Scopus subject areas

  • Biochemistry

Cite this

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title = "Increased spontaneous chemiluminescence from liver homogenates and isolated hepatocytes upon inhibition of o2 - and h2o2 utilization",
abstract = "The intracellular steady-state concentrations of hydrogen peroxide or Superoxide anion were increased by inhibiting either catalase, glutathione peroxidase, or Superoxide dismutase activities. Catalase was inhibited with aminotriazole while glutathione peroxidase activity was blocked by eliminating reduced glutathione after addition of either iodoacetamide diethylmaleate or phorone. The concentration of aminotriazole that stimulated chemiluminescence in 50{\%} (60 mM) was very similar to the Ki for catalase activity (70 mM). Cyanide, an inhibitor of both catalase and Superoxide dismutase, stimulated chemiluminescence in 50{\%} at a concentration (0.15 mM) which is much closer from the Ki for Superoxide dismutase (0.25 mM) than from the Ki for catalase (15 μM). The Superoxide dismutase inhibitor diethyldithiocarbamate also increased chemiluminescence six- to ten-fold. Depletion of reduced glutathione stimulated spontaneous chemiluminescence when its concentration decreased below 4.5 μmol · g liver-1. The results shown herein suggest that the changes in the intracellular steady-state concentration occurring after inhibition of any antioxidant enzyme are responsible for the increased spontaneous chemilumi-nescence. Spontaneous chemiluminescence from intact cells may be used as a noninvasive method for monitoring intracellular free radical metabolism.",
keywords = "Aminotriazole, Antioxidant enzymes, Catalase, Chemiluminescence, Glutathione peroxidase, Lipid peroxidation, Superoxide dismutase",
author = "Turrens, {Julio F.} and Giulivi, {Cecilia R} and Alberto Boveris",
year = "1986",
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TY - JOUR

T1 - Increased spontaneous chemiluminescence from liver homogenates and isolated hepatocytes upon inhibition of o2 - and h2o2 utilization

AU - Turrens, Julio F.

AU - Giulivi, Cecilia R

AU - Boveris, Alberto

PY - 1986

Y1 - 1986

N2 - The intracellular steady-state concentrations of hydrogen peroxide or Superoxide anion were increased by inhibiting either catalase, glutathione peroxidase, or Superoxide dismutase activities. Catalase was inhibited with aminotriazole while glutathione peroxidase activity was blocked by eliminating reduced glutathione after addition of either iodoacetamide diethylmaleate or phorone. The concentration of aminotriazole that stimulated chemiluminescence in 50% (60 mM) was very similar to the Ki for catalase activity (70 mM). Cyanide, an inhibitor of both catalase and Superoxide dismutase, stimulated chemiluminescence in 50% at a concentration (0.15 mM) which is much closer from the Ki for Superoxide dismutase (0.25 mM) than from the Ki for catalase (15 μM). The Superoxide dismutase inhibitor diethyldithiocarbamate also increased chemiluminescence six- to ten-fold. Depletion of reduced glutathione stimulated spontaneous chemiluminescence when its concentration decreased below 4.5 μmol · g liver-1. The results shown herein suggest that the changes in the intracellular steady-state concentration occurring after inhibition of any antioxidant enzyme are responsible for the increased spontaneous chemilumi-nescence. Spontaneous chemiluminescence from intact cells may be used as a noninvasive method for monitoring intracellular free radical metabolism.

AB - The intracellular steady-state concentrations of hydrogen peroxide or Superoxide anion were increased by inhibiting either catalase, glutathione peroxidase, or Superoxide dismutase activities. Catalase was inhibited with aminotriazole while glutathione peroxidase activity was blocked by eliminating reduced glutathione after addition of either iodoacetamide diethylmaleate or phorone. The concentration of aminotriazole that stimulated chemiluminescence in 50% (60 mM) was very similar to the Ki for catalase activity (70 mM). Cyanide, an inhibitor of both catalase and Superoxide dismutase, stimulated chemiluminescence in 50% at a concentration (0.15 mM) which is much closer from the Ki for Superoxide dismutase (0.25 mM) than from the Ki for catalase (15 μM). The Superoxide dismutase inhibitor diethyldithiocarbamate also increased chemiluminescence six- to ten-fold. Depletion of reduced glutathione stimulated spontaneous chemiluminescence when its concentration decreased below 4.5 μmol · g liver-1. The results shown herein suggest that the changes in the intracellular steady-state concentration occurring after inhibition of any antioxidant enzyme are responsible for the increased spontaneous chemilumi-nescence. Spontaneous chemiluminescence from intact cells may be used as a noninvasive method for monitoring intracellular free radical metabolism.

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KW - Glutathione peroxidase

KW - Lipid peroxidation

KW - Superoxide dismutase

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