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J Clin Invest. 1992 November; 90(5): 1871–1878.
PMCID: PMC443248

Identification of rifampin-inducible P450IIIA4 (CYP3A4) in human small bowel enterocytes.

Abstract

Enzymes within the P450IIIA (CYP3A) subfamily appear to account for significant "first pass" metabolism of some drugs in the intestine. To identify which of the known P450IIIA genes are expressed in intestine, enterocyte RNA was hybridized on Northern blots with synthetic oligonucleotides complementary to hypervariable regions of hepatic P450IIIA4, P450IIIA5, and P450IIIA7 cDNAs. Hybridization was detected only with the P450IIIA4-specific oligonucleotide. The identity of the hybridizing mRNA was confirmed to be P450IIIA4 by direct sequencing of a DNA fragment amplified from enterocyte cDNA by the polymerase chain reaction. To determine if enterocyte P450IIIA4 is inducible, biopsies of small bowel mucosa were obtained from five volunteers before and after they received 7d of treatment with rifampin, a known inducer of P450IIIA4 in liver. Rifampin treatment resulted in a five- or eightfold mean increase (P < 0.05) in the biopsy concentration of P450IIIA4 mRNA when normalized for content of sucrase isomaltase or intestinal fatty acid binding protein mRNAs, respectively. Rifampin also induced P450IIIA immunoreactive protein in enterocytes in each of the subjects, as judged by immunohistochemistry, and resulted in a 10-fold increase in P450IIIA4-specific catalytic activity (erythromycin N-demethylation) in the one patient studied. Our identification of inducible P450IIIA4 in enterocytes may in part account for drug interactions characteristic of P450IIIA4 substrates and suggests a strategy for controlling entry into the body of a major class of xenobiotics.

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Selected References

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  • Watkins PB, Wrighton SA, Maurel P, Schuetz EG, Mendez-Picon G, Parker GA, Guzelian PS. Identification of an inducible form of cytochrome P-450 in human liver. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6310–6314. [PubMed]
  • Shimada T, Guengerich FP. Evidence for cytochrome P-450NF, the nifedipine oxidase, being the principal enzyme involved in the bioactivation of aflatoxins in human liver. Proc Natl Acad Sci U S A. 1989 Jan;86(2):462–465. [PubMed]
  • Kronbach T, Fischer V, Meyer UA. Cyclosporine metabolism in human liver: identification of a cytochrome P-450III gene family as the major cyclosporine-metabolizing enzyme explains interactions of cyclosporine with other drugs. Clin Pharmacol Ther. 1988 Jun;43(6):630–635. [PubMed]
  • Combalbert J, Fabre I, Fabre G, Dalet I, Derancourt J, Cano JP, Maurel P. Metabolism of cyclosporin A. IV. Purification and identification of the rifampicin-inducible human liver cytochrome P-450 (cyclosporin A oxidase) as a product of P450IIIA gene subfamily. Drug Metab Dispos. 1989 Mar-Apr;17(2):197–207. [PubMed]
  • Gonzalez FJ, Schmid BJ, Umeno M, Mcbride OW, Hardwick JP, Meyer UA, Gelboin HV, Idle JR. Human P450PCN1: sequence, chromosome localization, and direct evidence through cDNA expression that P450PCN1 is nifedipine oxidase. DNA. 1988 Mar;7(2):79–86. [PubMed]
  • Guengerich FP, Martin MV, Beaune PH, Kremers P, Wolff T, Waxman DJ. Characterization of rat and human liver microsomal cytochrome P-450 forms involved in nifedipine oxidation, a prototype for genetic polymorphism in oxidative drug metabolism. J Biol Chem. 1986 Apr 15;261(11):5051–5060. [PubMed]
  • Bargetzi MJ, Aoyama T, Gonzalez FJ, Meyer UA. Lidocaine metabolism in human liver microsomes by cytochrome P450IIIA4. Clin Pharmacol Ther. 1989 Nov;46(5):521–527. [PubMed]
  • Guengerich FP, Müller-Enoch D, Blair IA. Oxidation of quinidine by human liver cytochrome P-450. Mol Pharmacol. 1986 Sep;30(3):287–295. [PubMed]
  • Kronbach T, Mathys D, Umeno M, Gonzalez FJ, Meyer UA. Oxidation of midazolam and triazolam by human liver cytochrome P450IIIA4. Mol Pharmacol. 1989 Jul;36(1):89–96. [PubMed]
  • Wang RW, Kari PH, Lu AY, Thomas PE, Guengerich FP, Vyas KP. Biotransformation of lovastatin. IV. Identification of cytochrome P450 3A proteins as the major enzymes responsible for the oxidative metabolism of lovastatin in rat and human liver microsomes. Arch Biochem Biophys. 1991 Nov 1;290(2):355–361. [PubMed]
  • Jacolot F, Simon I, Dreano Y, Beaune P, Riche C, Berthou F. Identification of the cytochrome P450 IIIA family as the enzymes involved in the N-demethylation of tamoxifen in human liver microsomes. Biochem Pharmacol. 1991 Jun 15;41(12):1911–1919. [PubMed]
  • Nebert DW, Nelson DR, Coon MJ, Estabrook RW, Feyereisen R, Fujii-Kuriyama Y, Gonzalez FJ, Guengerich FP, Gunsalus IC, Johnson EF, et al. The P450 superfamily: update on new sequences, gene mapping, and recommended nomenclature. DNA Cell Biol. 1991 Jan-Feb;10(1):1–14. [PubMed]
  • Molowa DT, Schuetz EG, Wrighton SA, Watkins PB, Kremers P, Mendez-Picon G, Parker GA, Guzelian PS. Complete cDNA sequence of a cytochrome P-450 inducible by glucocorticoids in human liver. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5311–5315. [PubMed]
  • Aoyama T, Yamano S, Waxman DJ, Lapenson DP, Meyer UA, Fischer V, Tyndale R, Inaba T, Kalow W, Gelboin HV, et al. Cytochrome P-450 hPCN3, a novel cytochrome P-450 IIIA gene product that is differentially expressed in adult human liver. cDNA and deduced amino acid sequence and distinct specificities of cDNA-expressed hPCN1 and hPCN3 for the metabolism of steroid hormones and cyclosporine. J Biol Chem. 1989 Jun 25;264(18):10388–10395. [PubMed]
  • Wrighton SA, Brian WR, Sari MA, Iwasaki M, Guengerich FP, Raucy JL, Molowa DT, Vandenbranden M. Studies on the expression and metabolic capabilities of human liver cytochrome P450IIIA5 (HLp3). Mol Pharmacol. 1990 Aug;38(2):207–213. [PubMed]
  • Bork RW, Muto T, Beaune PH, Srivastava PK, Lloyd RS, Guengerich FP. Characterization of mRNA species related to human liver cytochrome P-450 nifedipine oxidase and the regulation of catalytic activity. J Biol Chem. 1989 Jan 15;264(2):910–919. [PubMed]
  • Guengerich FP. Characterization of human microsomal cytochrome P-450 enzymes. Annu Rev Pharmacol Toxicol. 1989;29:241–264. [PubMed]
  • Watkins PB. Role of cytochromes P450 in drug metabolism and hepatotoxicity. Semin Liver Dis. 1990 Nov;10(4):235–250. [PubMed]
  • Watkins PB, Murray SA, Winkelman LG, Heuman DM, Wrighton SA, Guzelian PS. Erythromycin breath test as an assay of glucocorticoid-inducible liver cytochromes P-450. Studies in rats and patients. J Clin Invest. 1989 Feb;83(2):688–697. [PMC free article] [PubMed]
  • Pichard L, Fabre I, Fabre G, Domergue J, Saint Aubert B, Mourad G, Maurel P. Cyclosporin A drug interactions. Screening for inducers and inhibitors of cytochrome P-450 (cyclosporin A oxidase) in primary cultures of human hepatocytes and in liver microsomes. Drug Metab Dispos. 1990 Sep-Oct;18(5):595–606. [PubMed]
  • Ged C, Rouillon JM, Pichard L, Combalbert J, Bressot N, Bories P, Michel H, Beaune P, Maurel P. The increase in urinary excretion of 6 beta-hydroxycortisol as a marker of human hepatic cytochrome P450IIIA induction. Br J Clin Pharmacol. 1989 Oct;28(4):373–387. [PMC free article] [PubMed]
  • Wrighton SA, Ring BJ, Watkins PB, VandenBranden M. Identification of a polymorphically expressed member of the human cytochrome P-450III family. Mol Pharmacol. 1989 Jul;36(1):97–105. [PubMed]
  • Gupta SK, Bakran A, Johnson RW, Rowland M. Cyclosporin-erythromycin interaction in renal transplant patients. Br J Clin Pharmacol. 1989 Apr;27(4):475–481. [PMC free article] [PubMed]
  • First MR, Schroeder TJ, Weiskittel P, Myre SA, Alexander JW, Pesce AJ. Concomitant administration of cyclosporin and ketoconazole in renal transplant recipients. Lancet. 1989 Nov 18;2(8673):1198–1201. [PubMed]
  • Daniels NJ, Dover JS, Schachter RK. Interaction between cyclosporin and rifampicin. Lancet. 1984 Sep 15;2(8403):639–639. [PubMed]
  • Watkins PB, Hamilton TA, Annesley TM, Ellis CN, Kolars JC, Voorhees JJ. The erythromycin breath test as a predictor of cyclosporine blood levels. Clin Pharmacol Ther. 1990 Aug;48(2):120–129. [PubMed]
  • Kolars JC, Awni WM, Merion RM, Watkins PB. First-pass metabolism of cyclosporin by the gut. Lancet. 1991 Dec 14;338(8781):1488–1490. [PubMed]
  • Kolars JC, Stetson PL, Rush BD, Ruwart MJ, Schmiedlin-Ren P, Duell EA, Voorhees JJ, Watkins PB. Cyclosporine metabolism by P450IIIA in rat enterocytes--another determinant of oral bioavailability? Transplantation. 1992 Mar;53(3):596–602. [PubMed]
  • Watkins PB, Wrighton SA, Schuetz EG, Molowa DT, Guzelian PS. Identification of glucocorticoid-inducible cytochromes P-450 in the intestinal mucosa of rats and man. J Clin Invest. 1987 Oct;80(4):1029–1036. [PMC free article] [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Traber PG, Yu L, Wu GD, Judge TA. Sucrase-isomaltase gene expression along crypt-villus axis of human small intestine is regulated at level of mRNA abundance. Am J Physiol. 1992 Jan;262(1 Pt 1):G123–G130. [PubMed]
  • Komori M, Nishio K, Ohi H, Kitada M, Kamataki T. Molecular cloning and sequence analysis of cDNA containing the entire coding region for human fetal liver cytochrome P-450. J Biochem. 1989 Feb;105(2):161–163. [PubMed]
  • Sweetser DA, Birkenmeier EH, Klisak IJ, Zollman S, Sparkes RS, Mohandas T, Lusis AJ, Gordon JI. The human and rodent intestinal fatty acid binding protein genes. A comparative analysis of their structure, expression, and linkage relationships. J Biol Chem. 1987 Nov 25;262(33):16060–16071. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Dahlqvist A. Assay of intestinal disaccharidases. Scand J Clin Lab Invest. 1984 Apr;44(2):169–172. [PubMed]
  • NASH T. The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochem J. 1953 Oct;55(3):416–421. [PubMed]
  • Wrighton SA, Maurel P, Schuetz EG, Watkins PB, Young B, Guzelian PS. Identification of the cytochrome P-450 induced by macrolide antibiotics in rat liver as the glucocorticoid responsive cytochrome P-450p. Biochemistry. 1985 Apr 23;24(9):2171–2178. [PubMed]
  • Poland AP, Nebert DW. A sensitive radiometric assay of aminopyrine N-demethylation. J Pharmacol Exp Ther. 1973 Jan;184(1):269–277. [PubMed]
  • Beaune PH, Umbenhauer DR, Bork RW, Lloyd RS, Guengerich FP. Isolation and sequence determination of a cDNA clone related to human cytochrome P-450 nifedipine oxidase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8064–8068. [PubMed]
  • Schuetz JD, Molowa DT, Guzelian PS. Characterization of a cDNA encoding a new member of the glucocorticoid-responsive cytochromes P450 in human liver. Arch Biochem Biophys. 1989 Nov 1;274(2):355–365. [PubMed]
  • Komori M, Nishio K, Kitada M, Shiramatsu K, Muroya K, Soma M, Nagashima K, Kamataki T. Fetus-specific expression of a form of cytochrome P-450 in human livers. Biochemistry. 1990 May 8;29(18):4430–4433. [PubMed]
  • Murray GI, Barnes TS, Sewell HF, Ewen SW, Melvin WT, Burke MD. The immunocytochemical localisation and distribution of cytochrome P-450 in normal human hepatic and extrahepatic tissues with a monoclonal antibody to human cytochrome P-450. Br J Clin Pharmacol. 1988 Apr;25(4):465–475. [PMC free article] [PubMed]
  • Ptachcinski RJ, Venkataramanan R, Burckart GJ. Clinical pharmacokinetics of cyclosporin. Clin Pharmacokinet. 1986 Mar-Apr;11(2):107–132. [PubMed]
  • Lucey MR, Kolars JC, Merion RM, Campbell DA, Aldrich M, Watkins PB. Cyclosporin toxicity at therapeutic blood levels and cytochrome P-450 IIIA. Lancet. 1990 Jan 6;335(8680):11–15. [PubMed]
  • Kolars JC, Schmiedlin-Ren P, Dobbins WO, 3rd, Schuetz J, Wrighton SA, Watkins PB. Heterogeneity of cytochrome P450IIIA expression in rat gut epithelia. Gastroenterology. 1992 Apr;102(4 Pt 1):1186–1198. [PubMed]

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