Reactive oxygen species and non-peroxidative mechanisms of cocaine-induced cytotoxicity in rat hepatocyte cultures.
Primary short-term cultures of hepatocytes derived from phenobarbital-induced male Sprague-Dawley rats were used to investigate the mechanisms of cocaine-induced cytotoxicity. Exposure of cells to cocaine resulted in a time and concentration-dependent release of lactate dehydrogenase (LDH) into the culture medium which became evident after 7 h of incubation. Over the course of 24 h incubation with cocaine (0.3 mM) there was no significant lipid peroxidation (measured as the formation of thiobarbituric acid-reactive substances. TBA-RS). The addition of the ferric iron chelator, deferoxamine (DFO), prevented in part cocaine-induced LDH release. Alternatively, addition of the antioxidant, alpha-tocopherol polyethylene glycol 1000 succinate (TPGS), did not protect against hepatocyte injury. Depletion of the intracellular glutathione (GSH) with diethyl maleate (DEM) to below critical levels for antioxidative protection markedly accelerated the onset and increased the extent of cocaine-induced LDH release, concomitant with massive production of lipid peroxidation. During the first four hours of incubation DFO and TPGS protected against cocaine-induced cytotoxicity in GSH-depleted cells. However, at later stages (24 h), the protective effect was lost even in the absence of lipid peroxidation. These results suggest that reactive oxygen species are involved in cocaine-mediated hepatocyte injury. However, lipid peroxidation can be dissociated from other, non-peroxidative, iron-dependent mechanisms of oxidative cell injury.
Institute of Toxicology, Swiss Federal Institute of Technology, Schwerzenbach.
C R
CR
Göldlin
U A
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Boelsterli
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003042:Cocaine / Q000633:toxicity*; D003676:Deferoxamine / Q000494:pharmacology; D004347:Drug Interactions; D005978:Glutathione / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D015227:Lipid Peroxidation; D008099:Liver / Q000166:cytology / Q000187:drug effects*; D008297:Male; D008298:Maleates / Q000633:toxicity; D010100:Oxygen / Q000378:metabolism*; D011092:Polyethylene Glycols; D051381:Rats; D011919:Rats, Inbred Strains; D014810:Vitamin E / Q000031:analogs & derivatives / Q000494:pharmacology
D008298:Maleates; D014810:Vitamin E; D011092:Polyethylene Glycols; D007770:L-Lactate Dehydrogenase; C014476:diethyl maleate; D005978:Glutathione; D003042:Cocaine; D003676:Deferoxamine; C014225:tocophersolan; D010100:Oxygen