H M Wortelboer, C A de Kruif, W I de Boer, A A van Iersel, H E Falke, B J Blaauboer
{"title":"Induction and activity of several isoenzymes of cytochrome P-450 in primary cultures of rat hepatocytes, in comparison with in vivo data.","authors":"H M Wortelboer, C A de Kruif, W I de Boer, A A van Iersel, H E Falke, B J Blaauboer","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>Changes in cytochrome P-450 isoenzymes were studied in rat liver and in primary cultures of rat hepatocytes after treatment with compounds belonging to various classes of inducers, including phenobarbital (PB), beta-naphthoflavone (BNF), and clofibrate/clofibric acid (CLOF/CLOFA). The enzyme activity toward specific substrates was measured, and the presence of apoprotein of several P-450 isoenzymes was determined semiquantitatively by Western blotting. In untreated cultures the P-450 content and activities of 7-ethoxyresorufin O-deethylation (EROD) and aniline 4-hydroxylation (AH) declined with time at different rates. In cultures treated with BNF, the protein levels of isoenzyme P-450IA1 and P-450IA2 were elevated, as in vivo. This induction was reflected in a markedly increased EROD activity. CLOFA enhanced the AH and EROD activity in primary cultures at the same level as in vivo. The monooxygenase activity pentoxyresorufin O-depentylation (PROD) was stimulated by PB and CLOF in vivo, which correlated with the enhanced protein level of P-450IIB1/2. In contrast, the PROD activity was not induced when cultures were treated with PB or CLOFA, although we could detect apoprotein of P-450IIB1/2 by immunoblotting.</p>","PeriodicalId":77750,"journal":{"name":"Molecular toxicology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1987-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular toxicology","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
Changes in cytochrome P-450 isoenzymes were studied in rat liver and in primary cultures of rat hepatocytes after treatment with compounds belonging to various classes of inducers, including phenobarbital (PB), beta-naphthoflavone (BNF), and clofibrate/clofibric acid (CLOF/CLOFA). The enzyme activity toward specific substrates was measured, and the presence of apoprotein of several P-450 isoenzymes was determined semiquantitatively by Western blotting. In untreated cultures the P-450 content and activities of 7-ethoxyresorufin O-deethylation (EROD) and aniline 4-hydroxylation (AH) declined with time at different rates. In cultures treated with BNF, the protein levels of isoenzyme P-450IA1 and P-450IA2 were elevated, as in vivo. This induction was reflected in a markedly increased EROD activity. CLOFA enhanced the AH and EROD activity in primary cultures at the same level as in vivo. The monooxygenase activity pentoxyresorufin O-depentylation (PROD) was stimulated by PB and CLOF in vivo, which correlated with the enhanced protein level of P-450IIB1/2. In contrast, the PROD activity was not induced when cultures were treated with PB or CLOFA, although we could detect apoprotein of P-450IIB1/2 by immunoblotting.