Glutathione modulates toxic oxygen metabolite injury of canine chief cell monolayers in primary culture.
Cultured canine gastric chief cells exposed to a toxic oxygen metabolite-generating system (xanthine plus xanthine oxidase) demonstrated minimal cytolysis, suggesting that these cells have important endogenous antioxidant mechanisms. We have quantified the role of glutathione for protection against toxic oxygen metabolites by measuring cell lysis by lactate dehydrogenase release after variable depletion and repletion of cellular glutathione content. In the absence of exogenous oxidant stress, the glutathione content of chief cells can be depleted to less than 0.2 nmol total glutathione/micrograms DNA or 22% of control without cell lysis over 5 h. However, when challenged with the oxygen metabolite-generating system, cytolysis was greatly enhanced by glutathione depletion. Oxygen metabolite-mediated cytolysis after glutathione depletion was inhibited by exogenous catalase, thiourea, and deferoximine, but not superoxide dismutase or mannitol. These data suggested that hydrogen peroxide and hydroxyl radical mediated cytolysis in glutathione-depleted chief cells. If a substrate for glutathione synthesis, N-acetyl-L-cysteine, was provided to the depleted cells for 1 h before challenge with the oxygen radical-generating system, cell lysis was markedly decreased. However, if glutathione synthesis was blocked during the repletion period by buthionine sulfoximine, protection was not restored. The data supported an important role for glutathione as an endogenous antioxidant, which modulated the sensitivity of cultured chief cells to toxic oxygen metabolite injury.
Olson
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CE
Center for Ulcer Research and Education, Wadsworth Veterans Administration Medical Center, Los Angeles, California.
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002478:Cells, Cultured; D003584:Cytological Techniques; D003603:Cytotoxins / Q000494:pharmacology; D004285:Dogs; D004789:Enzyme Activation; D005609:Free Radicals; D005753:Gastric Mucosa / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism / Q000494:pharmacology*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008298:Maleates / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism*; D013270:Stomach / Q000166:cytology / Q000187:drug effects*; D013997:Time Factors; D019820:Xanthine; D014969:Xanthine Oxidase / Q000378:metabolism / Q000494:pharmacology; D014970:Xanthines / Q000494:pharmacology
D000975:Antioxidants; D003603:Cytotoxins; D005609:Free Radicals; D008298:Maleates; D014970:Xanthines; D019820:Xanthine; D006861:Hydrogen Peroxide; D014969:Xanthine Oxidase; C014476:diethyl maleate; D005978:Glutathione; D010100:Oxygen