Chao Y S, Pickett C B, Yamin T T, Guo L S, Alberts A W, Kroon P A
Mol Pharmacol. 1985 Mar;27(3):394-8.
The effect of phenobarbital on the level of rat liver apolipoprotein A-I (apo-A-I) mRNA was studied. Poly(A+)-RNA isolated from livers of control or phenobarbital-treated rats was translated in vitro in the rabbit reticulocyte lysate system and immunoprecipitated with rabbit antiserum against rat apo-A-I. The immunoprecipitate was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The translational activity of apo-A-I mRNA was estimated from the incorporation of [35S]methionine into the apo-A-I band. It was found to be elevated 4-fold by 16 hr after rats received a single injection of phenobarbital. To study the effect of phenobarbital on the level of rat liver apo-A-I mRNA, a recombinant plasmid which contained a cDNA insert corresponding to rat liver apo-A-I mRNA was isolated and used to hybridize total liver poly(A+)-RNA from control and phenobarbital-treated rats. There were 4.8- and 10-fold increases in the amount of hybridization to mRNAs from rats after they were treated with phenobarbital for 8 and 16 hr, respectively. Thus, phenobarbital increases the level of rat liver apo-A-I mRNA.
研究了苯巴比妥对大鼠肝脏载脂蛋白A-I（apo-A-I）mRNA水平的影响。从对照或苯巴比妥处理的大鼠肝脏中分离的多聚腺苷酸（Poly(A+)）-RNA在兔网织红细胞裂解物系统中进行体外翻译，并用抗大鼠apo-A-I的兔抗血清进行免疫沉淀。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析免疫沉淀物。根据[35S]甲硫氨酸掺入apo-A-I条带的情况估计apo-A-I mRNA的翻译活性。发现大鼠单次注射苯巴比妥16小时后，其翻译活性提高了4倍。为了研究苯巴比妥对大鼠肝脏apo-A-I mRNA水平的影响，分离了一个含有与大鼠肝脏apo-A-I mRNA对应的cDNA插入片段的重组质粒，并用于与对照和苯巴比妥处理的大鼠肝脏总多聚腺苷酸（Poly(A+)）-RNA杂交。大鼠经苯巴比妥处理8小时和16小时后其mRNA杂交量分别增加了4.8倍和10倍。因此，苯巴比妥可提高大鼠肝脏apo-A-I mRNA的水平。
