Abstract
The predominant inducible cytochrome P450 (CYP) in rat small intestine is CYP1A1, which, when induced to elevated levels by xenobiotics or dietary constituents, has the potential to metabolize and consequently reduce the systemic uptake of low concentrations of orally ingested, bioactivatable polycyclic aromatic hydrocarbons and heterocyclic aromatic amines. We investigated the regulation of small intestinal CYP1A1 in an effort to develop its anticancer potential. The time courses of hepatic and intestinal CYP1A1 induction by β-naphthoflavone (BNF) were compared quantitatively at the protein and mRNA levels by immunoblot and competitive RNA-polymerase chain reaction analyses. CYP1A1 mRNA levels in both organs increased sharply and were maximal at ∼6 hr and returned to near basal levels by 12 hr after BNF treatment. In contrast, hepatic CYP1A2 mRNA levels increased much more gradually. Small intestinal CYP1A2 mRNA concentrations were insufficient to support translation of detectable protein. Maximal levels of intestinal and hepatic CYP1A1 protein occurred between 12 and 24 hr, and 24 and 48 hr, respectively, after BNF. Intestinal CYP1A1 protein was detectable earlier and for a shorter duration than hepatic CYP1A1. CYP1A1 induction was first detected in crypt cells 3 hr before the appearance of activity in villous cells, and maximal levels of activity were reached in crypt cells 12 to 18 hr before maximal and 1.5-fold (per mg protein) higher responses in villous cells—induction thus occurs in both villous and crypt cells. Previously detected decreases in CYP1A1 inducibility from duodenum to ileum correlated with decreases in immunoblot determined-Ah receptor levels. Intestinal CYP1A1 induction does not involve the glucocorticoid receptor in contrast to hepatic induction. These studies have revealed several novel features of small intestinal CYP1A1 regulation.
Footnotes
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Send reprint requests to: Dr. Laurence S. Kaminsky, New York State Department of Health, Wadsworth Center, P.O. Box 509, Empire State Plaza, Albany, NY 12201-0509.
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This research was funded by Grant ES06256 from the National Institutes of Health, Public Health Service.
- Abbreviations used are::
- P450
- cytochrome P450
- CYP
- cytochrome P450
- BNF
- β-naphthoflavone
- PAH
- polycyclic aromatic hydrocarbon
- DEX
- dexamethasone
- IgG
- immunoglobulin G
- BCA
- bicinchoninic acid
- ECL
- enhanced chemiluminescence
- PCR
- polymerase chain reaction
- bp
- base pair
- SDS-PAGE
- sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- AhR
- arylhydrocarbon receptor
- TBST
- 20 mM Tris HCl (pH 7.4), containing 0.5 M NaCl and 0.05% Tween-20
- Received July 22, 1996.
- Accepted October 8, 1996.
- The American Society for Pharmacology and Experimental Therapeutics
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