Toxic consequence of the abrupt depletion of glutathione in cultured rat hepatocytes.
Cultured hepatocytes were exposed to two chemicals, dinitrofluorobenzene (DNFB) and diethyl maleate (DEM), that abruptly deplete cellular stores of glutathione. Upon the loss of GSH, lipid peroxidation was evidenced by an accumulation of malondialdehyde in the cultures followed by the death of the hepatocytes. Pretreatment of the hepatocytes with a ferric iron chelator, deferoxamine, or the addition of an antioxidant, N,N'-diphenyl-p-phenylenediamine (DPPD), to the culture medium prevented both the lipid peroxidation and the cell death produced by either DNFB or DEM. However, neither deferoxamine nor DPPD prevented the depletion of GSH caused by either agent. Inhibition of glutathione reductase by 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) or inhibition of catalase by aminotriazole sensitized the hepatocytes to the cytotoxicity of DNFB. In a similar manner, pretreatment with BCNU potentiated the cell killing by DEM. DPPD and deferoxamine protected hepatocytes pretreated with BCNU and then exposed to DNFB or DEM. These data indicate that an abrupt depletion of GSH leads to lipid peroxidation and cell death in cultured hepatocytes. It is proposed that GSH depletion sensitizes the hepatocyte to its constitutive flux of partially reduced oxygen species. Such an oxidative stress is normally detoxified by GSH-dependent mechanisms. However, with GSH depletion these activated oxygen species are toxic as a result of the iron-dependent formation of a potent oxidizing species.
Department of Pathology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
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S
Miccadei
M E
ME
Kyle
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D
Gilfor
J L
JL
Farber
D000640:Amitrole / Q000494:pharmacology; D000818:Animals; D002330:Carmustine / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004139:Dinitrofluorobenzene / Q000633:toxicity*; D004347:Drug Interactions; D005978:Glutathione / Q000378:metabolism*; D008054:Lipid Peroxides / Q000096:biosynthesis; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008298:Maleates / Q000633:toxicity*; D009578:Nitrobenzenes / Q000633:toxicity*; D010084:Oxidation-Reduction / Q000187:drug effects; D051381:Rats; D011919:Rats, Inbred Strains
D008054:Lipid Peroxides; D008298:Maleates; D009578:Nitrobenzenes; D004139:Dinitrofluorobenzene; C014476:diethyl maleate; D005978:Glutathione; D002330:Carmustine; D000640:Amitrole