Effects of ethanol and clofibrate on expression of cytochrome P-450 enzymes and epoxide hydrolase in cultures and cocultures of rat hepatocytes.
Cultured and cocultured rat hepatocytes were used to study the effects of ethanol and clofibrate on cytochrome P-450 (P-450) enzymes and epoxide hydrolase. We showed that in the presence of ethanol, clofibrate or both compounds, rat hepatocytes were able to express, after 3 days of pure culture, quantitatively and qualitatively reasonable levels of most cytochrome P-450 enzymes and epoxide hydrolase, compared to freshly isolated hepatocytes. However, ethanol induced the P-450IA subfamily, and clofibrate the P-450-IVA subfamily. In cocultures, after 6 days, most P-450 enzymes were still expressed while P-450IIC11 was completely lost. Ethanol and clofibrate had the same effect as in pure culture. These results show, by modifying culture medium conditions and cell-cell interactions, that it is possible to maintain reasonable xenobiotic-metabolizing-enzyme expression; however, these conditions have to be improved in order to preserve better P-450 expression. The mechanism of these effects and the inducibility of these systems remain to be elucidated by a study at molecular level.
Perrot
N
N
Institut National de la Santé et de la Recherche Médicale Unité 75, Centre Hospitalo-Universitaire Necker-Enfants-Malades, Paris, France.
Chesné
C
C
De Waziers
I
I
Conner
J
J
Beaune
P H
PH
Guillouzo
A
A
D000818:Animals; D002478:Cells, Cultured; D002994:Clofibrate / Q000494:pharmacology*; D003470:Culture Media; D046508:Culture Techniques; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004851:Epoxide Hydrolases / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology*; D015151:Immunoblotting; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains
D003470:Culture Media; D000431:Ethanol; D003577:Cytochrome P-450 Enzyme System; D004851:Epoxide Hydrolases; D002994:Clofibrate