3,3'-Diindolylmethane induces CYP1A2 in cultured precision-cut human liver slices.

1. The effect of 3,3'-diindolylmethane (DIM), an indole derivative derived from cruciferous vegetables, on cytochrome P450 (CYP) isoforms in the CYP1A and CYP3A subfamilies has been studied in 72-h cultured human liver slices. 2. In cultured human liver slices 50 microM DIM induced 7-ethoxyresorufin O-deethylase and to a lesser extent 7-methoxyresorufin O-demethylase activities. 3. Western immunoblotting of liver slice microsomes was performed with antibodies to rat CYP1A2 and human CYP3A4. Compared with control liver slice microsomes (dimethyl sulphoxide-only treated), DIM induced levels of CYP1A2 but had little effect on levels of CYP3A4. The treatment of human liver slices with 2 microg/ml of the polycholorinated biphenyl mixture Aroclor 1254 also resulted in an induction of levels of CYP1A2, but had no effect on CYP3A4. 4. These results demonstrate that DIM induces CYP1A isoforms in cultured human liver slices. Some variability in the magnitude of induction of enzyme activities by DIM was observed in four human liver samples examined. For 7-ethoxyresorufin O-deethylase, the magnitude of induction by 50 microM DIM ranged from 2.3- to 19.3-fold. 5. These results demonstrate that cultured human liver slices can be used to evaluate the effect of chemicals derived from cruciferous and other vegetables on CYP isoforms.

[1]  J. Tredger,et al.  Induction of CYP3A isoforms in cultured precision-cut human liver slices. , 1997, Xenobiotica; the fate of foreign compounds in biological systems.

[2]  S. Kitareewan,et al.  Validation of precision-cut liver slices in dynamic organ culture as an in vitro model for studying CYP1A1 and CYP1A2 induction. , 1996, Toxicology and applied pharmacology.

[3]  A. Okey.  Enzyme induction in the cytochrome P-450 system. , 1990, Pharmacology & therapeutics.

[4]  A. B. Hanley,et al.  Chemical and biological properties of indole glucosinolates (glucobrassicins): a review. , 1988, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[5]  R. J. Price,et al.  Culture of precision-cut liver slices: effect of some peroxisome proliferators. , 1993, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[6]  J. Yager,et al.  Cytochrome P450 isoenzyme activities in cultured rat and mouse liver slices. , 1997, Xenobiotica; the fate of foreign compounds in biological systems.

[7]  Andrew Parkinson,et al.  Strategies for restoration and maintenance of normal hepatic structure and function in long-term cultures of rat hepatocytes , 1996 .

[8]  A. Conney,et al.  Stimulatory effect of brussels sprouts and cabbage on human drug metabolism , 1979, Clinical pharmacology and therapeutics.

[9]  R. McDanell,et al.  The Effect of Brassica Vegetable Consumption on Caffeine Metabolism in Humans , 1992, Human & experimental toxicology.

[10]  C. Cho,et al.  A major inducer of anticarcinogenic protective enzymes from broccoli: isolation and elucidation of structure. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[11]  A. Conney,et al.  Effect of brussels sprouts and cabbage on drug conjugation , 1984, Clinical pharmacology and therapeutics.

[12]  R. Edwards,et al.  Identification of the epitope of an anti-peptide antibody which binds to CYP1A2 in many species including man. , 1993, Biochemical pharmacology.

[13]  P. Maurel.  The use of adult human hepatocytes in primary culture and other in vitro systems to investigate drug metabolism in man , 1996 .

[14]  J. Domergue,et al.  Omeprazole and lansoprazole are mixed inducers of CYP1A and CYP3A in human hepatocytes in primary culture. , 1994, The Journal of pharmacology and experimental therapeutics.

[15]  D. Müller,et al.  Monooxygenation, cytochrome P4501A1 and P4501A1-mRNA in rat liver slices exposed to beta-naphthoflavone and dexamethasone in vitro. , 1996, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[16]  S. Thorgeirsson,et al.  Contribution of CYP1A1 and CYP1A2 to the activation of heterocyclic amines in monkeys and human. , 1994, Carcinogenesis.

[17]  C. Bonfils,et al.  Induction protocols for cytochromes P450IIIA in vivo and in primary cultures of animal and human hepatocytes. , 1991, Methods in enzymology.

[18]  J. Noordhoek,et al.  Acid reaction products of indole-3-carbinol and their effects on cytochrome P450 and phase II enzymes in rat and monkey hepatocytes. , 1992, Biochemical pharmacology.

[19]  A. Conney.  Induction of microsomal cytochrome P-450 enzymes: the first Bernard B. Brodie lecture at Pennsylvania State University. , 1986, Life sciences.

[20]  J. Noordhoek,et al.  Structure elucidation of acid reaction products of indole-3-carbinol: detection in vivo and enzyme induction in vitro. , 1991, Chemico-biological interactions.

[21]  J. Noordhoek,et al.  Effects of indole-3-carbinol on biotransformation enzymes in the rat: in vivo changes in liver and small intestinal mucosa in comparison with primary hepatocyte cultures. , 1992, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[22]  S. Loft,et al.  Foreign compound metabolism capacity in man measured from metabolites of dietary caffeine. , 1992, Carcinogenesis.

[23]  J. Tredger,et al.  Induction of cytochrome P450 isoenzymes in cultured precision-cut rat and human liver slices. , 1996, Xenobiotica; the fate of foreign compounds in biological systems.

[24]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[25]  J. Clausen,et al.  Effects of dietary broccoli on human in vivo drug metabolizing enzymes: evaluation of caffeine, oestrone and chlorzoxazone metabolism. , 1996, Carcinogenesis.

[26]  D W Nebert,et al.  P450 superfamily: update on new sequences, gene mapping, accession numbers and nomenclature. , 1996, Pharmacogenetics.

[27]  R. J. Price,et al.  Induction of cytochrome P-450-dependent enzyme activities in cultured rat liver slices. , 1993, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[28]  A. B. Hanley,et al.  Differential induction of mixed-function oxidase (MFO) activity in rat liver and intestine by diets containing processed cabbage: correlation with cabbage levels of glucosinolates and glucosinolate hydrolysis products. , 1987, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.