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 Scopus citations

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

Keywords

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

ASJC Scopus subject areas

  • Biochemistry

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