Buthionine sulfoximine protects the viability of adult rat Leydig cells exposed to ethane dimethanesulfonate.
Exposure to (EDS) in vivo or in vitro alters adult rat Leydig cell function within 3 hr and these cells subsequently die within 24-48 hr. Previously, we determined that reducing intracellular glutathione levels with buthionine sulfoximine (BSO) protects adult rat Leydig cell function during short-term (3 hr) EDS exposures. The present study extends these observations by assessing whether BSO also can prevent Leydig cell death following EDS exposure. To assess whether BSO can prevent EDS-induced Leydig cell death, Leydig cells were cultured in the presence or the absence of 4 mM BSO (12 hr), and subsequently with increasing doses of EDS (3 hr). The effects of EDS on Leydig cell death were estimated using a cytotoxicity assay (MTT assay) and by assessing the ability of adult rat Leydig cells to maintain LH-stimulated testosterone production and [35S]methionine incorporation 24 hr following EDS exposure. This culture duration was chosen because sustained alterations in MTT reduction, LH-stimulated testosterone production, and [35S]methionine incorporation 24 hr following EDS exposure were found to reflect alterations in Leydig cell viability. The results suggest that lowering intracellular levels of glutathione with BSO prevents EDS-induced Leydig cell death. Restoring intracellular levels of glutathione with glutathione ethyl ester (8 mM) in the presence of BSO also restored the ability of EDS to kill Leydig cells. Taken together, these data suggest that glutathione mediates the mechanism by which EDS kills adult rat Leydig cells.
Kelce
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ManTech Environmental Technology, Inc., Research Triangle Park, North Carolina 27709.
D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology*; D019328:Buthionine Sulfoximine; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D007985:Leydig Cells / Q000187:drug effects*; D008297:Male; D008698:Mesylates / Q000633:toxicity*; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives* / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley
D000963:Antimetabolites; D008698:Mesylates; D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; C002994:ethylene dimethanesulfonate