title: Immunochemical detection of cytochrome P450 isozymes induced in rat liver by n-hexane, 2-hexanone and acetonyl acetone.
abstract: Cytochrome P450 isozymes induced in rat liver by treatment with n-hexane, 2-hexanone and acetonyl acetone (given intraperitoneally 5 mmol/kg for 4 days) were investigated using enzyme assays (benzene, toluene, 7-ethoxyresorufin and 7-pentoxyresorufin metabolism) and monoclonal antibodies (anti-P450IA1/2, anti-P450IIB1/2, anti-P450IIC11/6, anti-P450IIE1(91) and anti-P450IIE1(98)). n-Hexane treatment enhanced the activities of low-Km benzene aromatic hydroxylase and toluene side-chain oxidase, but not 7-ethoxyresorufin O-deethylase or 7-pentoxyresorufin O-depentylase. 2-Hexanone or acetonyl acetone treatment enhanced the activities of low- and high-Km benzene aromatic hydroxylases, toluene side-chain oxidase and 7-pentoxyresorufin O-depentylase, but not of 7-ethoxyresorufin O-deethylase. Immunoblot analysis showed that anti-P450IA1/2 did not bind liver microsomal protein from either control and treated rats in the region of cytochrome P450s, whereas with anti-P450IIE1(98) a clear-cut band was seen in liver microsomes from control and treated rats, with intensities in the following order: 2-hexanone = acetonyl acetone greater than or equal to n-hexane greater than control greater than phenobarbital. With anti-P450IIB1/2, a band was detected in microsomes from phenobarbital-treated rats, and to a lesser extent, in microsomes from 2-hexanone- and acetonyl acetone-treated rats. Like the immunoblot analysis, anti-P450IIE1(91) inhibited toluene side-chain hydroxylase activity in all microsomes, except in preparations from phenobarbital-treated rats and anti-P450IIB1 in microsomes from phenobarbital-, 2-hexanone- and acetonyl acetone-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Hygiene, Shinshu University School of Medicine, Matsumoto, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nakajima
authors.forename: E
authors.initials: E
authors.lastname: Elovaara
authors.forename: S S
authors.initials: SS
authors.lastname: Park
authors.forename: H V
authors.initials: HV
authors.lastname: Gelboin
authors.forename: H
authors.initials: H
authors.lastname: Vainio
mesh_terms: D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis* / Q000096:biosynthesis; D004790:Enzyme Induction / Q000187:drug effects; D006586:Hexanes / Q000494:pharmacology*; D006588:Hexanones / Q000633:toxicity*; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D007527:Isoenzymes / Q000032:analysis* / Q000096:biosynthesis; D008297:Male; D008742:Methyl n-Butyl Ketone / Q000494:pharmacology*; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D006586:Hexanes; D006588:Hexanones; D007527:Isoenzymes; D008742:Methyl n-Butyl Ketone; D003577:Cytochrome P-450 Enzyme System; C011269:2,5-hexanedione