Cytotoxicity of mitomycin C and adriamycin in freshly isolated rat hepatocytes: the role of cytochrome P450.
The role of cytochrome P450 (P450) in the cytotoxicity of mitomycin C (MMC) and Adriamycin (ADR) was investigated in freshly isolated hepatocytes from phenobarbital-induced rats. The loss of cell viability [measured as lactate dehydrogenase (LDH) leakage] upon MMC exposure was accompanied by a rapid and extensive intracellular glutathione (GSH) depletion and followed by minor lipid peroxidation (LPO). Coincubation of the hepatocytes with the P450 inhibitors, metyrapone and SK&F 525-A, strongly protected against MMC-induced LDH leakage, GSH depletion, and LPO. Inasmuch as the depletion of intracellular GSH by MMC, which is considered as a critical event in the development of MMC cytotoxicity, was not accompanied by a stoichiometric oxidation to oxidized GSH (GSSG), the formation of a MMC-GSH conjugate after one-electron reductive bioactivation of MMC by P450 was anticipated. In contrast to MMC, ADR was only cytotoxic to the hepatocytes upon prior depletion of intracellular GSH with diethylmaleate. Addition of metyrapone and SK&F 525-A completely protected the hepatocytes against ADR-induced LDH leakage and LPO. Moreover, the ADR-induced LDH leakage and LPO were strongly inhibited by dimethyl sulfoxide and ethanol, indicating that hydroxyl radicals were involved in the cytotoxicity of ADR. In conclusion, the present investigations indicate that P450 plays a major role in the cytotoxicity of both MMC and ADR in freshly isolated hepatocytes from phenobarbital-induced rats. The present findings lead to a better understanding of the mechanism of the cytotoxic actions of both MMC and ADR.
Goeptar
A R
AR
Division of Molecular Toxicology, Leiden/Amsterdam Center for Drug Research, Vrije Universiteit, Amsterdam, The Netherlands.
Groot
E J
EJ
Scheerens
H
H
Commandeur
J N
JN
Vermeulen
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NP
D000818:Animals; D003374:Coumarins / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism / Q000502:physiology*; D004317:Doxorubicin / 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* / Q000378:metabolism; D008297:Male; D008298:Maleates / Q000494:pharmacology; D008797:Metyrapone / Q000494:pharmacology; D016685:Mitomycin / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D011335:Proadifen / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
D003374:Coumarins; D008298:Maleates; C017299:7-ethoxycoumarin; D016685:Mitomycin; D004317:Doxorubicin; D003577:Cytochrome P-450 Enzyme System; D011335:Proadifen; D007770:L-Lactate Dehydrogenase; C014476:diethyl maleate; D005978:Glutathione; D008797:Metyrapone