不同诱导方法制备大鼠肝微粒体酶及其活性的比较

Comparison of different induction methods on activity of rat liver microsomal enzymes

目的:通过比较4种方法诱导制备的大鼠肝微粒体酶(S9)的蛋白含量及活化效果,探讨有效的肝微粒体酶诱导方法。方法:分别采用多氯联苯、苯巴比妥联合β-萘黄酮、注射用苯巴比妥钠联合β-萘黄酮、苯巴比妥作为诱导剂制备大鼠肝S9。采用Lowry法测定S9蛋白含量,通过Ames试验和中国仓鼠肺成纤维细胞(CHL细胞)体外染色体畸变试验对4种S9的活化效果进行比较。结果:上述4种方法制备的S9蛋白含量依次为30.29、34.15、33.31、28.02 mg/m L,均不超过40 mg/m L。Ames试验和染色体畸变试验中,与不加S9组比较,4种S9在阳性组中均显示出明显的活化作用(P<0.01)。前3种S9所得阳性结果无明显差异,以苯巴比妥作为单独诱导剂的活化作用较前三者弱,差异具有统计学意义(P<0.05或P<0.01)。结论:在本试验条件和范围下,以多氯联苯、苯巴比妥联合β-萘黄酮、注射用苯巴比妥钠联合β-萘黄酮作为诱导剂可以成功制备大鼠肝S9,并具有较好的活性,联合诱导法可替代多氯联苯诱导法。

OBJECTIVE： To compare different induction methods on activity of rat liver microsomal enzyme S9and to identify the effective induction method. METHODS：Rats were injected with PCBs,phenobarbital,betanaphthoflavoneand phenobarbitol sodium. In addition,beta-naphthoflavone and phenobarbital were used as inducer toinduce rat liver S9. S9 protein content was determined by the method of Lowry. Metabolic activation effects of four kinds ofS9 were compared using the Ames test and the in vitro chromosome aberration assay of CHL cells. RESULTS：Proteincontents of four kinds of S9 were 30.29,34.15,33.31,28.02 mg/mL,all of them were less than 40 mg/mL. Comparedwith the groups without S9,the 4 types of S9 showed activation effect in the positive control （P〈0.01）. Activation effectsfrom three kinds of S9 had no significant difference among them in the Ames and CHL chromosome aberration assays（P〉0.05）,however,the activation from the phenobarbital-induced S9 was significantly weaker （P〈0.05 or P〈0.01）.CONCLUSION：Under our conditions：using PCBs,phenobarbital and beta-naphthoflavone,phenobarbitol sodiuminjection and beta-naphthoflavonen as inducer,metabolically active rat liver S9 was induced successfully. In addition,the combined induction method can take the place of the induction by PCBs alone.