肝细胞色素P4501A1、2E1在非酒精性脂肪肝发病中的作用

The role of hepatocyte cytochrome P450 1A1, 2E1 in rat nonalcoholic steatosis

目的 研究肝细胞色素P4 5 0 1A1、2E1在非酒精性脂肪肝大鼠中的表达及其意义。方法 荧光分光光度计法检测高脂饲料诱导 4 0只大鼠脂肪肝模型中P4 5 0 1A1活性 (7 乙氧异唑O 脱乙基酶 ,EROD)和 2E1活性 (苯胺羟化酶 ,ANH) ,免疫组化和Westernblot方法测定肝细胞色素P4 5 0 1A1、2E1表达 ,逆转录聚合酶链反应测定其mRNA表达。结果 脂肪肝 2、4、8和 1 2周EROD活性分别为(32 5 .0 7± 5 9.6 8)、(345 .2 5± 4 9.2 8)、(4 6 8.95± 5 5 .2 8)和 (5 4 8.6 8± 4 3.2 5 )nmol·mg-1 ·min-1 ,与正常对照组的 (2 6 0 .4 2± 35 .32 )nmol·mg-1 ·min-1 比较明显增高 (P <0 .0 1 ) ,脂肪肝 2、4、8和 1 2周ANH活性分别为 (6 35 .6 8± 6 5 .4 8)、(735 .4 5± 76 .89)、(887.4 5± 85 .6 5 )和 (95 6 .5 8± 84 .4 7)nmol·mg-1 ·min-1 ,与正常对照组的 (5 0 0 .2 5± 78.34)nmol·mg-1 ·min-1 比较明显增高 (P <0 .0 1 ) ,肝细胞色素P4 5 0 1A1、2E1基因表达随脂肪肝程度的加重而增高。结论 脂肪肝引起的肝脏损害可能与肝细胞色素P4 5 0 1A1、2E1表达有关 。

Objective To detect the expression of hepatocyte cytochrome P450 1A1,2E1 in rat non alcoholic steatosis model. Methods 7 ethoxyresorufin O deethylase (EROD) and benzopyrene hydroxylase (ANH) activity were determined by ultraviolet chromatography. The expression of hepatocyte cytochrome P450 1A1, 2E1 protein in the liver of rats with non alcoholic steatosis induced by high fat diet was detected with immunohistochemistry and Western blot. Cytochrome P450 1A1, 2E1 mRNA were assayed with reverse transcriptase polymerase chain reaction(RT PCR). Results The levels of EROD activity in nonalcoholic steatosis rat at 2, 4, 8, 12 week and normal control group were (325.07±59.68),(345.25±49.28),( 468.95 ±55.28),( 548.68 ±43.25) and (260.42±35.32) nmol·mg -1 ·min -1 respectively. The levels of ANH activity in nonalcoholic steatosis rat at 2, 4, 8, 12 week and normal control group were (635.68 ±65.48), (735.45 ±76.89 ),(887.45±85.65),(956.58±84.47) and (500.25±78.34) nmol·mg -1 ·min -1 respectively. These indicate that the levels of EROD and ANH activity in rats with nonalcoholic steatosis were significantly increased compared with that in control group. In addition, the expression of hepatocyte cytochrome P450 1A1, 2E1 protein and cytochrome P450 1A1, 2E1 mRNA in rats with nonalcoholic steatosis were significantly higher than that in control group. The level of P450 1A1 and 2E1 was increased correspondingly with the degree of nonalcoholic steatosis. Conclusions The induction of hepatocyte cytochrome P4501A1 and 2E1 might participate in the development of nonalcoholic steatosis