Induction of cytochromes P450 2B and 2E1 in rat liver by isomeric picoline N-oxides.
Pyridine derivatives are widely used solvents and precursors for the synthesis of chemicals of industrial importance. Oxidized metabolites have been implicated in the observed toxicity of pyridines and are known to induce drug-metabolizing enzymes in rat liver. In this study the three isomeric picoline (methylpyridine) N-oxides, as major oxidized metabolites of 2-, 3- and 4-picoline, were evaluated as inducers of cytochrome P450 (CYP) enzymes in rat liver. After a single dose of 100 mg/kg 24 h before sacrifice the 3- and 4-isomers were effective inducers of microsomal substrate oxidations associated with the phenobarbital-inducible CYPs 2B; upregulation of CYP2B protein was confirmed by immunoblotting. In contrast, the 2-isomer did not increase CYP2B protein or activity in rat liver but CYP2E1 protein expression was upregulated by the isomers to 160-200% of control. The three chemicals increased aniline 4-hydroxylation activity in rat liver, which is consistent with induction of CYPs 2B or 2E1 and 4-nitrophenol 2-hydroxylation activity was increased in microsomal fractions from 3- and 4-picoline N-oxide-treated rats. The activities of several other CYPs were also determined and CYP1A-dependent 7-ethylresorufin O-deethylation was increased (to approximately 6- and 2-fold of control) by the 3- and 4-isomer, respectively, whereas the activity of CYP3A-mediated androstenedione 6beta-hydroxylation was decreased by the agents--most notably by the 2-isomer. During NADPH-supported oxidation of CCl4, lipid peroxidation was increased in microsomes from 3- and 4-picoline N-oxide-pretreated rats and was modulated in vitro by the CYP2B inhibitor orphenadrine, but not by the CYP2E1 inhibitor 4-methylpyrazole. These findings establish that particular isomers of picoline N-oxide rapidly upregulate CYP2B or, to a lesser extent, CYP2E1 and implicate CYP2B in the enhanced lipid peroxidation observed in microsomes from rats treated with 3- and 4-picoline N-oxides. Such induction process may contribute to the hepatotoxicity of pyridines by enhancing the capacity for microsomal lipid peroxidation.
Murray
M
M
Department of Medicine, University of Sydney, Westmead Hospital, NSW, Australia. M.Murray@hri.edu.au
Sefton
R M
RM
Martini
R
R
Butler
A M
AM
D000818:Animals; D019362:Cytochrome P-450 CYP2B1 / Q000096:biosynthesis* / Q000187:drug effects; D019392:Cytochrome P-450 CYP2E1 / Q000096:biosynthesis* / Q000187:drug effects; D004790:Enzyme Induction; D007536:Isomerism; D015227:Lipid Peroxidation; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D009569:Nitric Oxide / Q000378:metabolism / Q000494:pharmacology; D010847:Picolines / Q000378:metabolism* / Q000494:pharmacology / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar
D010847:Picolines; D009569:Nitric Oxide; D019392:Cytochrome P-450 CYP2E1; D019362:Cytochrome P-450 CYP2B1