Toftgård R, Nilsen O G, Ingelman-Sundberg M, Gustafsson J A
Acta Pharmacol Toxicol (Copenh). 1980 May;46(5):353-61. doi: 10.1111/j.1600-0773.1980.tb02466.x.
Multiple forms of liver microsomal cytochrome P-450 in rats were identified on SDS-polyacrylamide gels stained for protein and peroxidase activity after induction with phenobarbital, 3-methylcholanthrene, and 16 alpha-cyanopregnenolone. The induced forms were correlated to the in vitro metabolism of biphenyl, benzo(a)pyrene and the steroids 4-androstene-3,17-dione and 5 alpha-androstane-3 alpha,17 beta-diol. Induction of two forms with apparent molecular weights of 54,000 (RLvMc P-450 54) AND 50,000 (RLvMc P-450 50) was obtained with phenobarbital, induction of RLvMc P-450 55 and RLvMc P-450 58 with 3-methylcholanthrene and induction of FLvMc P-450 54 with 16 alpha-cyanopregnenolone. The RLvMc P-450 50 was mainly associated with the formation of benzo(a)pyrene-4,5-dihydrodiol, and 7 alpha-hydroxy-4-androstene-3,17-dione. The RLvMc P-450 55 and/or the RLvMc P-450 58 was mainly associated with the formation of 2- and 3-hydroxybiphenyl and benzo(a)pyrene-7,8-dihydrodiol and FLvMc P-450 54 was to some extent associated with the formation of benzo(a)pyrene-4,5-dihydrodiol and several metabolites of 5 alpha-androstane-3 alpha, 17 beta-diol. It is suggested that SDS-polyacrylamide gel electrophoresis may be a valuable complement to enzyme assays in evaluating effects of drugs and environmental chemicals on the liver microsomal hydroxylase system.
在用苯巴比妥、3-甲基胆蒽和16α-氰基孕烯醇酮诱导后,通过对蛋白质和过氧化物酶活性进行染色的SDS-聚丙烯酰胺凝胶,鉴定出大鼠肝脏微粒体细胞色素P-450的多种形式。诱导出的形式与联苯、苯并(a)芘以及甾体4-雄烯-3,17-二酮和5α-雄甾烷-3α,17β-二醇的体外代谢相关。用苯巴比妥诱导出两种表观分子量分别为54,000(RLvMc P-450 54)和50,000(RLvMc P-450 50)的形式,用3-甲基胆蒽诱导出RLvMc P-450 55和RLvMc P-450 58,用16α-氰基孕烯醇酮诱导出FLvMc P-450 54。RLvMc P-450 50主要与苯并(a)芘-4,5-二氢二醇以及7α-羟基-4-雄烯-3,17-二酮的形成有关。RLvMc P-450 55和/或RLvMc P-450 58主要与2-和3-羟基联苯以及苯并(a)芘-7,8-二氢二醇的形成有关,而FLvMc P-450 54在一定程度上与苯并(a)芘-4,5-二氢二醇以及5α-雄甾烷-3α,17β-二醇的几种代谢产物的形成有关。有人提出,SDS-聚丙烯酰胺凝胶电泳在评估药物和环境化学物质对肝脏微粒体羟化酶系统的影响时,可能是酶分析的一种有价值的补充。