Chlormethiazole as an efficient inhibitor of cytochrome P450 2E1 expression in rat liver.
Ethanol-inducible cytochrome P450 (CYP) 2E1 (CYP2E1) is responsible for the metabolism of many xenobiotics which exert toxic effects in humans. Specific inhibitors might constitute valuable tools in the elucidation of the pharmacological and toxicological roles of this isozyme in vivo. In the present investigation we have evaluated the effects of a drug used for treatment of ethanol withdrawal states, chloromethiazole (CMZ), on CYP2E1 expression in rat liver. A 4-fold induction of CYP2E1 was observed after 3 days of starvation, accompanied by a similar increase in the level of the corresponding mRNA. CMZ specifically inhibited the elevation of CYP2E1 mRNA and protein, but did not prevent CYP2B1 and CYP3A1 or CYP1A1 induction caused by treatment with phenobarbital or beta-naphthoflavone, respectively. From nuclear run-off experiments it was apparent that the rate of the CYP2E1 gene transcription was inhibited greatly by CMZ treatment. Rats treated with ethanol in a total enteral nutrition model had higher CYP2E1-dependent hepatic microsomal activities of p-nitrophenol hydroxylase and carbon tetrachloride-induced lipid peroxidation than controls, and simultaneous CMZ treatment abolished the ethanol-dependent induction. In vitro experiments with rat liver microsomes showed that CMZ did not act as an inhibitor of CYP2E1-dependent catalytic activities or as an inhibitor of microsomal NADPH and CYP2E1-dependent lipid peroxidation. In conclusion, we suggest that CMZ might constitute an efficient and specific inhibitor of CYP2E1 expression suitable for in vivo experiments.
Hu
Y
Y
Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden.
Mishin
V
V
Johansson
I
I
von Bahr
C
C
Cross
A
A
Ronis
M J
MJ
Badger
T M
TM
Ingelman-Sundberg
M
M
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D002719:Chlormethiazole / Q000378:metabolism / Q000494:pharmacology*; D019392:Cytochrome P-450 CYP2E1; D051544:Cytochrome P-450 CYP3A; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D008297:Male; D010089:Oxidoreductases, N-Demethylating / Q000037:antagonists & inhibitors* / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley
D065607:Cytochrome P-450 Enzyme Inhibitors; D002719:Chlormethiazole; D003577:Cytochrome P-450 Enzyme System; D019392:Cytochrome P-450 CYP2E1; D001189:Aryl Hydrocarbon Hydroxylases; C465138:Cyp3a23-3a1 protein, rat; D051544:Cytochrome P-450 CYP3A; D010089:Oxidoreductases, N-Demethylating