Activities of drug metabolizing enzymes in bovine colon epithelial cell cultures
Autor: | Angelika Dohle, H.M. Bolt, Günter Schmahl, Susanne Weber, Wolfram Föllmann, Sascha Birkner |
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Rok vydání: | 2003 |
Předmět: |
Sulfotransferase
Colon Health Toxicology and Mutagenesis Prostaglandin Toxicology Mixed Function Oxygenases Xenobiotics chemistry.chemical_compound Cytosol Animals RNA Messenger Cells Cultured chemistry.chemical_classification biology Reverse Transcriptase Polymerase Chain Reaction Cytochrome P450 Epithelial Cells General Medicine Metabolism Glutathione Immunohistochemistry Culture Media Rats Microscopy Electron Enzyme Pharmaceutical Preparations chemistry Biochemistry Cell culture Microscopy Electron Scanning Microsomes Liver biology.protein Cattle Biomarkers Drug metabolism |
Zdroj: | Archives of Toxicology. 77:621-629 |
ISSN: | 1432-0738 0340-5761 |
Popis: | The metabolic competence of cultured bovine colon epithelial cells was evaluated by determining activities of phase I and II enzymes in colonocytes cultured for different intervals (maximum of 10 days) compared with activities measured in freshly isolated cells. Cytochrome p50 1A1-associated 7-ethoxyresorufin O-deethylase (EROD) activity was detectable in freshly isolated colonocytes and in colon cells maintained in culture for up to 5 days. In contrast to liver samples, cytochrome p50 3A4-associated 7-benzyloxyresorufin O-debenzylase (BROD) activity was not detectable in bovine colon cells. Prostaglandin H synthase-mediated production of prostaglandin E(2) was found in freshly isolated and also in cultured colonocytes. Both isoenzymes (COX 1 and COX 2) were detected in cultured cells. To examine phase II metabolic potency, activities of N-acetyltransferases 1 and 2, of phenol and amino sulfotransferases, of glutathione S-transferases alpha, mu, pi and theta and of UDP-glucuronyltransferase were measured. N-Acetyltransferase (NAT) activity (substrate p-aminobenzoic acid, PABA, a diagnostic substrate for the human NAT-1 enzyme) was stable under culture conditions and during the observed culture period comparable to that of freshly isolated cells. In contrast, sulfamethazine, a specific substrate for NAT-2, was not acetylated, neither in bovine colon cells nor in bovine liver samples. Whereas activity of amino sulfotransferase (substrate 2-naphthylamine) decreased continuously during the entire culture period, the activity of phenol sulfotransferase (substrate 1-naphthol) decreased only slowly. Activity of total glutathione S-transferases (alpha, mu, and pi) (substrate 1-chloro-2,4-dinitrobenzene) decreased after 2 days in culture, but was stable during the following culture period. Activity of glutathione S-transferase theta (substrate epoxy-3-nitrophenoxypropane) changed during the culture period. At the beginning and the end (after 10 days) of the culture period maximum activity was measured. Activity of UDP-glucuronyltransferase increased during the culture period reaching a maximum after 7 days. The results show that cultured bovine epithelial colon cells express several enzyme activities required for the biotransformation of xenobiotics. |
Databáze: | OpenAIRE |
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