Title
Comparison of coumarin-induced toxicity between sandwich-cultured primary rat hepatocytes and rats in vivo: A toxicogenomics approach
Author
Kienhuis, A.S.
Wortelboer, H.M.
Hoflack, J.C.
Moonen, E.J.
Kleinjans, J.C.S.
van Ommen, B.
van Delft, J.H.M.
Stierum, R.H.
TNO Kwaliteit van Leven
Publication year
2006
Abstract
Sandwich-cultured primary rat hepatocytes are often used as an in vitro model in toxicology and pharmacology. However, loss of liver-specific functions, in particular, the decline of cytochrome P450 (P450) enzyme activity, limits the value of this model for prediction of in vivo toxicity. In this study, we investigated whether a hepatic in vitro system with improved metabolic competence enhances the predictability for coumarin-induced in vivo toxicity by using a toxicogenomics approach. Therefore, primary rat hepatocytes were cultured in sandwich configuration in medium containing a mixture of low concentrations of P450 inducers, phenobarbital, dexamethasone, and β-naphthoflavone. The toxicogenomics approach used enabled comparison of similar mechanistic endpoints at the molecular level between in vitro and in vivo conditions, namely, compound-induced changes in multiple genes and signaling pathways. Toxicant-induced cytotoxic effects and gene expression profiles observed in hepatocytes cultured in modified medium and hepatocytes cultured in standard medium (without inducers) were compared with results from a rat in vivo study. Coumarin was used as a model compound because its toxicity depends on bioactivation by P450 enzymes. Metabolism of coumarin toward active metabolites, coumarin-induced cytotoxicity, and gene expression modulation were more pronounced in hepatocytes cultured in modified medium compared with hepatocytes cultured in standard medium. In addition, more genes and biological pathways were similarly affected by coumarin in hepatocytes cultured in modified medium and in vivo. In conclusion, these experiments showed that for coumarin-induced toxicity, sandwich-cultured hepatocytes maintained in modified medium better represent the situation in vivo compared with hepatocytes cultured in standard medium. Copyright © 2006 by The American Society for Pharmacology and Experimental Therapeutics.
Subject
Biology
Biomedical Research
beta naphthoflavone
coumarin
cytochrome P450
dexamethasone
drug metabolite
enzyme inducing agent
phenobarbital
animal cell
animal experiment
animal model
animal tissue
article
controlled study
culture medium
cytotoxicity
drug activation
drug metabolism
enzyme activity
experimental rat
gene expression
histopathology
in vitro study
in vivo study
liver cell
liver toxicity
male
nonhuman
nucleotide sequence
prediction
principal component analysis
priority journal
rat
signal transduction
toxicogenetics
Alanine Transaminase
Animals
Aspartate Aminotransferases
Cell Culture Techniques
Cells, Cultured
Cholesterol
Coumarins
gamma-Glutamyltransferase
Gene Expression Profiling
Hepatocytes
Liver
Male
Oligonucleotide Array Sequence Analysis
Rats
Rats, Wistar
Toxicogenetics
To reference this document use:
http://resolver.tudelft.nl/uuid:3eea1fec-79cc-4614-a6b5-0d42049fbc97
DOI
https://doi.org/10.1124/dmd.106.011262
TNO identifier
239699
ISSN
0090-9556
Source
Drug Metabolism and Disposition, 34 (12), 2083-2090
Document type
article