Effects of the antifungal imidazole ketoconazole on CYP1A and CYP3A in rainbow trout and killifish.
The use of N-substituted imidazoles is widespread, and imidazole and triazole fungicides have been detected in the aquatic environment and shown to bioaccumulate in fish. We have investigated effects of the model imidazole, ketoconazole, on drug-metabolizing cytochrome P450 (CYP) forms. We focused on cytochrome P4501A (CYP1A) and cytochrome P4503A (CYP3A) expression and activities in juvenile rainbow trout and in adult killifish. The CYP1A expression (mRNA, protein) and activity was induced in rainbow trout, whereas in killifish no effect of ketoconazole on CYP1A protein expression was observed. A biphasic dose-response relationship was observed between ketoconazole exposure and hepatic CYP1A-mediated ethoxyresorufin O-deethylase (EROD) activity in rainbow trout in vitro and in vivo, implying that higher doses of ketoconazole inhibit CYP1A activities. Slight induction of CYP3A protein levels was observed in rainbow trout exposed in vivo to ketoconazole. However, the CYP3A-mediated benzyloxy-4-[trifluoromethyl]-coumarin (BFC) O-debenzyloxylase activity was reduced in rainbow trout and killifish treated with ketoconazole. In vitro inhibition studies confirmed that ketoconazole was a potent inhibitor of both CYP3A and CYP1A enzyme activities in these species. This study showed that ketoconazole induced CYP1A and CYP3A expression in rainbow trout. However, the most pronounced effect of ketoconazole was a 60 to 90% decrease in CYP3A catalytic activities in rainbow trout and in killifish.
Hegelund
Tove
T
Göteborg University, Department of Zoophysiology, Box 463, SE 405 30 Göteborg, Sweden.
Ottosson
Karin
K
Rådinger
Madeleine
M
Tomberg
Peter
P
Celander
Malin C
MC
D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D001189:Aryl Hydrocarbon Hydroxylases / Q000187:drug effects / Q000378:metabolism*; D019363:Cytochrome P-450 CYP1A1 / Q000187:drug effects / Q000378:metabolism*; D051544:Cytochrome P-450 CYP3A; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D023781:Fundulidae / Q000378:metabolism*; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology; D007413:Intestinal Mucosa / Q000378:metabolism; D007654:Ketoconazole / Q000494:pharmacology*; D008099:Liver / Q000378:metabolism; D017686:Oncorhynchus mykiss / Q000378:metabolism*; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism
D000935:Antifungal Agents; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D001189:Aryl Hydrocarbon Hydroxylases; D019363:Cytochrome P-450 CYP1A1; D051544:Cytochrome P-450 CYP3A; C106496:cytochrome P-450 CYP1A3 (rainbow trout); D010089:Oxidoreductases, N-Demethylating; D007654:Ketoconazole