Role of cellular thiol status in tocopheryl hemisuccinate cytoprotection against ethyl methanesulfonate-induced toxicity.
Suspensions of rat hepatocytes treated with the alkylating agent ethyl methanesulfonate (EMS) exhibited extensive lipid peroxidation as well as rapid and near complete depletion of cellular reduced glutathione (GSH) levels prior to cell death. Pretreatment of hepatocytes with medium deficient in sulfur amino acids accelerated cell death induced by EMS, confirming the previously reported cytoprotective role for GSH in this toxic event. Nearly all of the cellular GSH lost following 50 mM EMS treatment was accounted for as S-ethyl glutathione (GS-Et). No significant formation of glutathione disulfide was observed. The GS-Et formed was not exported from the cell but remained at high intracellular concentrations throughout the course of the experiment. In addition, EMS treatment inhibited the efflux of intracellular GSH and inhibited the cellular accumulation of glutamate (Glu). Supplementation of hepatocytes with 25 microM d-alpha-tocopheryl hemisuccinate (TS) protected these cells against EMS-induced lipid peroxidation and cell death. Cytoprotection with TS had no effect on EMS-induced depletion of intracellular GSH or intracellular levels of GS-Et or Glu. However, TS supplementation did prevent EMS-induced depletion of cellular protein thiols. Interestingly, the pretreatment of hepatocytes with 1 mM dithiothreitol promoted EMS toxicity. The results of this study suggest that the cytoprotective abilities of TS are related to the prevention of both EMS-induced lipid peroxidation and protein thiol depletion. Thus, the onset of lipid peroxidation and the loss of protein thiols in hepatocytes appear to be critical cellular events leading to EMS-induced cell death.
Fariss
M W
MW
Department of Pharmaceutical Sciences, College of Pharmacy, Washington State University, Pullman 99164-6510, USA.
Bryson
K F
KF
Tirmenstein
M A
MA
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D004229:Dithiothreitol / Q000494:pharmacology; D005020:Ethyl Methanesulfonate / Q000633:toxicity*; D005978:Glutathione / Q000502:physiology; D015227:Lipid Peroxidation / Q000187:drug effects; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D013438:Sulfhydryl Compounds / Q000502:physiology*; D024505:Tocopherols; D014810:Vitamin E / Q000031:analogs & derivatives* / Q000494:pharmacology
D000975:Antioxidants; D013438:Sulfhydryl Compounds; D014810:Vitamin E; D005020:Ethyl Methanesulfonate; D005978:Glutathione; D024505:Tocopherols; D004229:Dithiothreitol