Studies with nitrogen-containing steroids and freshly isolated rat hepatocytes: role of cytochrome P-450 in detoxication.
Several nitrogen-containing steroids produced concentration- and time-dependent decreases in the viability of freshly isolated F-344 rat hepatocytes. N,N-Diethyl-4-methyl-3-oxo-4-aza-5 alpha-androst-1-ene-17 beta-carboxamide (I) was not cytotoxic at or below 0.3 mM but produced decreases in cell viability at higher concentrations. In contrast, the desmethyl analog of I was essentially nontoxic, demonstrating that relatively small structural changes result in substantial differences in cytotoxicity. Testosterone and other steroids specifically potentiated the cytotoxicity of I in a concentration-dependent manner, while having no effect upon the toxicity of other chemical agents. Pargyline and methimazole had no effect upon the cytotoxicity of I, suggesting that monoamine oxidase and flavin-containing monooxygenase are not involved. The cytochrome P-450 inhibitors octylamine and metyrapone potentiated the cytotoxicity of I. Induction of cytochrome P-450 isozymes by phenobarbital and beta-naphthoflavone treatment protected the cells against the cytotoxicity of I, while acetone or dexamethasone treatment had no effect. The initial rates of hepatocyte metabolism of the six nitrogen-containing steroids investigated did not correlate with cytotoxicity. Dithiothreitol and other thiol compounds had no effect upon the cytotoxicity of I, suggesting that sulfhydryl oxidation is not involved. Galactosamine and sulfate-free media had no effect upon the cytotoxicity of I. These results suggest that cytochrome P-450 is involved in the detoxication of I by rat hepatocytes while conjugative metabolism does not play a significant role.
Kedderis
G L
GL
Department of Animal Drug Metabolism, Merck Sharp & Dohme Research Laboratories, Rahway, New Jersey 07065.
Argenbright
L S
LS
Miwa
G T
GT
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D003577:Cytochrome P-450 Enzyme System / Q000502:physiology*; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004790:Enzyme Induction / Q000187:drug effects; D005965:Glucuronates / Q000378:metabolism; D066298:In Vitro Techniques; D008658:Inactivation, Metabolic; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D013256:Steroids / Q000378:metabolism / Q000633:toxicity*; D013329:Structure-Activity Relationship; D013431:Sulfates / Q000378:metabolism; D013739:Testosterone / Q000633:toxicity
D005965:Glucuronates; D013256:Steroids; D013431:Sulfates; D013739:Testosterone; D003577:Cytochrome P-450 Enzyme System; D004229:Dithiothreitol