Induction of phase I and II drug metabolism in rat small intestine and colon in vitro.
The aim of this study was to evaluate drug metabolism in rat small intestinal and colon precision-cut slices during 24 h of incubation and the applicability of these slices for enzyme induction studies. Various parameters were evaluated: intracellular levels of ATP (general viability marker), alkaline phosphatase activity (specific epithelial marker), villin expression (specific epithelial marker), and metabolic rates of 7-ethoxycoumarin (CYP1A), testosterone (CYP3A and CYP2B), and 7-hydroxycoumarin (glucuronide and sulfate conjugation) conversions. ATP and villin remained constant up to, respectively, 5 and 8 h in small intestine and up to 24 h in colon. The metabolic rate remained constant in small intestinal slices up to 8 h and decreased afterward to 24 to 92%, depending on the substrate studied. The inducibility of metabolism in small intestinal and colon slices was tested with several inducers at various concentrations and incubation times. The following inducers were used: 3-methylcholanthrene, beta-naphthoflavone, indirubin, and tert-butylhydroquinone (aryl hydrocarbon receptor ligands), dexamethasone (glucocorticoid receptor/pregnane X receptor ligand) and phenobarbital (constitutive androstane receptor ligand). After incubation with inducers, metabolic rates were evaluated with 7-ethoxycoumarin and testosterone (phase I) and 7-hydroxycoumarin (phase II) as substrate. All inducers elevated the metabolic rates consistent with the available published in vivo induction data. Induction of enzyme activity was already detectable after 5 h (small intestine) and after 8 h (colon) for 3-methylcholanthrene and beta-naphthoflavone and was clearly detectable for all tested inducers after 24 h (up to 20-fold compared with noninduced controls). In conclusion, small intestinal and colon precision-cut slices are useful for metabolism and enzyme induction studies.
van de Kerkhof
E G
EG
Department of Pharmacokinetics and Drug Delivery, Gronigen University Institute for Drug Exploration, University of Groningen, The Netherlands. e.g.van.de.kerkhof@rug.nl
de Graaf
I A M
IA
de Jager
M H
MH
Groothuis
G M M
GM
D000255:Adenosine Triphosphate / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D003106:Colon / Q000378:metabolism*; D003374:Coumarins / Q000378:metabolism; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000235:genetics; D066298:In Vitro Techniques; D007421:Intestine, Small / Q000378:metabolism*; D008297:Male; D050216:Metabolic Detoxication, Phase I; D050217:Metabolic Detoxication, Phase II; D008840:Microfilament Proteins / Q000235:genetics; D010599:Pharmacokinetics; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013739:Testosterone / Q000378:metabolism; D014468:Umbelliferones / Q000378:metabolism
D003374:Coumarins; D008840:Microfilament Proteins; D012333:RNA, Messenger; D014468:Umbelliferones; C026615:villin; C017299:7-ethoxycoumarin; D013739:Testosterone; C031477:7-hydroxycoumarin; D000255:Adenosine Triphosphate; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+); D000469:Alkaline Phosphatase