title: Inhibition of CYP1A2 and CYP3A4 by oltipraz results in reduction of aflatoxin B1 metabolism in human hepatocytes in primary culture.
abstract: Dithiolethiones are thought to act as potent chemoprotective agents against aflatoxin B1 (AFB1)-induced hepatocarcinogenesis in the rat by inducing glutathione S-transferases (GSTs). To determine whether these antioxidants can be similarly effective in human beings, we have investigated metabolism of AFB1, in primary human hepatocytes with or without pretreatment by oltipraz (OPZ), a synthetic derivative of the natural 1,2-dithiole-3-thione. Aflatoxin M1 (AFM1), glutathione conjugates of AFB1 oxides (AFBSGs), and unchanged AFB1 were quantitated in cultures derived from eight human liver donors. Parenchymal cells obtained from the three GST M1-positive livers metabolized AFB1 to AFM1 and to AFBSGs derived from the isomeric exo-and endo-8,9-oxides, whereas no AFBSGs were formed in the GST M1-null cells. Pretreatment of the cells with 3-methylcholanthrene or rifampicin, inducers of CYP1A2 and CYP3A4, respectively, caused a significant increase in AFB1 metabolism. Although OPZ induced GST A2, and to a lesser extent GST A1 and GST M1, it decreased formation of AFM1 and AFBSG, which involves CYP1A2 and CYP3A4. Inhibition by OPZ of AFB1 metabolism by reducing CYP1A2 and CYP3A4 was also demonstrated by decreased activity of their monooxygenase activities toward ethoxyresorufin and nifedipine, respectively. The significant inhibition by OPZ of human recombinant yeast CYP1A2 and CYP3A4 was also shown. These results demonstrate that AFBSG can be formed by GST M1-positive human hepatocytes only, and suggest that chemoprotection with OPZ is due to an inhibition of activation of AFB1, in addition to a GST-dependent inactivation of the carcinogenic exo-epoxide.
authors.affiliation: INSERM U49, Centre Hospitalier Regional Universitaire Ponchaillou, Rennes, France.
authors.forename: S
authors.initials: S
authors.lastname: Langouët
authors.forename: B
authors.initials: B
authors.lastname: Coles
authors.forename: F
authors.initials: F
authors.lastname: Morel
authors.forename: L
authors.initials: L
authors.lastname: Becquemont
authors.forename: P
authors.initials: P
authors.lastname: Beaune
authors.forename: F P
authors.initials: FP
authors.lastname: Guengerich
authors.forename: B
authors.initials: B
authors.lastname: Ketterer
authors.forename: A
authors.initials: A
authors.lastname: Guillouzo
mesh_terms: D016604:Aflatoxin B1 / Q000378:metabolism*; D000368:Aged; D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D019388:Cytochrome P-450 CYP1A2; D051544:Cytochrome P-450 CYP3A; D065607:Cytochrome P-450 Enzyme Inhibitors; D005260:Female; D005982:Glutathione Transferase / Q000378:metabolism*; D006801:Humans; D007223:Infant; D008099:Liver / Q000378:metabolism*; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D008875:Middle Aged; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors*; D011719:Pyrazines / Q000494:pharmacology*; D012293:Rifampin / Q000494:pharmacology; D013871:Thiones; D013876:Thiophenes; D013997:Time Factors
chemical_list: D016588:Anticarcinogenic Agents; D065607:Cytochrome P-450 Enzyme Inhibitors; D011719:Pyrazines; D013871:Thiones; D013876:Thiophenes; D008748:Methylcholanthrene; C026209:oltipraz; D016604:Aflatoxin B1; D006899:Mixed Function Oxygenases; D010088:Oxidoreductases; C104464:CYP3A protein, human; D019388:Cytochrome P-450 CYP1A2; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D005982:Glutathione Transferase; D012293:Rifampin