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目的:观察补肾祛湿方含药血清对雌激素致大鼠胆汁淤积肝细胞胆汁酸转运蛋白MRP2与BSEP表达的作用,探讨补肾祛湿方治疗妊娠肝内胆汁淤积症可能的分子机制。方法:用1μmol/L的17β雌二醇干预大鼠原代肝细胞24 h诱导形成胆汁淤积模型后,加入补肾祛湿方含药血清处理24 h,放射免疫法测定培养液中总胆汁酸含量,提取细胞总RNA及蛋白,实时荧光定量PCR法和蛋白免疫印迹法检测MRP2、BSEP mRNA与蛋白表达水平。电子显微镜观察培养肝细胞超微结构变化。结果:补肾祛湿方含药血清可减轻肝细胞损伤,显著增加细胞培养上清总胆汁酸含量,P<0.01,并且可明显增加肝细胞MRP2、BSEP mRNA与蛋白表达水平,P<0.01。结论:补肾祛湿方治疗妊娠肝内胆汁淤积症的分子机制可能是通过改善肝细胞超微结构的损伤,增加肝细胞胆汁酸转运蛋白MRP2、BSEP mRNA与蛋白表达水平实现的。
Objective: To observe the effect of Bushen Qushi prescription serum on estrogen-induced cholestatic rat liver cholestatic bile acid transporter expression of MRP2 and BSEP to explore the possible mechanism of Bushen Qushi prescription in treatment of intrahepatic cholestasis of pregnancy. Methods: Primary rat hepatocytes were treated with 17μmol / L 17β-estradiol for 24 hours to induce cholestatic model. The rats were treated with Bushen Qushi Decoction for 24 hours, and the total bile acid content in the medium was determined by radioimmunoassay The total RNA and protein of cells were extracted, and the mRNA and protein levels of MRP2 and BSEP were detected by real-time fluorescence quantitative PCR and Western blotting. Observation of ultrastructural changes in cultured hepatocytes by electron microscopy. Results: Bushen Qushi Decoction contained serum can reduce hepatocellular injury, significantly increase the total bile acid content of the cell culture supernatant, P <0.01, and can significantly increase the mRNA and protein expression of MRP2, BSEP in hepatocytes, P <0.01. Conclusion: The molecular mechanism of Bushen Qushi Decoction in treating intrahepatic cholestasis of pregnancy may be through improving the damage of hepatocyte ultrastructure and increasing the expression of MRP2, BSEP mRNA and protein of hepatocytes.