酒精性肝病模型小鼠胆红素清除通路变化以及组成型雄烷受体的表达的研究

Research on the Bilirubin Clearance Pathway and the Expression of Constitutive Androstane Receptor in a Murine Model of Alcoholic Liver Disease

目的研究酒精性肝病模型小鼠胆红素清除通路变化以及胆红素代谢调控因子组成型雄烷受体的表达情况。方法采用Lieber-De Carli酒精饲料以及等卡热的对照饲料予小鼠自由饮食4周构建酒精性肝病小鼠模型。检测总红素、直接胆红素水平,采用Western blot法、RT-PCR、免疫荧光法分析胆红素摄取转运体OATP1a1、OATP1a4、OATP1b2,葡萄糖醛酸转移酶UGT1A1、泵出转运体MRP2以及胆红素代谢调控因子CAR的表达。结果慢性酒精摄入提高血浆胆红素水平,在转录及翻译水平诱导UGT1A1表达、下调MRP2以及OATP1a1。此外,酒精性肝病小鼠肝核蛋白CAR表达明显抑制而肝总蛋白CAR未见明显改变。结论慢性酒精摄入选择性抑制胆红素清除通路,伴随胆红素代谢调控因子CAR易位损伤。

Objective To investigate the bilirubin clearance pathway and the expression of constitutive androstane receptor（CAR） in a murine model of alcoholic liver disease. Methods Mice were fed Lieber-DeCarli ethanol diet or an isocaloric control diet for 4 weeks. The serum total bilirubin and direct bilirubin were tested.The expression of uptake transports OATP1a1,OATP1a4,OATP1b2, bilirubin glucuronidating UGT1A1,excretion pump MRP2 and Bilirubin metabolism regulation factor CAR were analyzed by western blot and quantitative RT-PCR assay, as well as Immunofluorescence microscopy. Results Chronic alcohol intake elevated plasma bilirubin levels accompanied by induction of UGT1A1, reduction of MRP2, OATP1a1 on transcriptional level and post-transcriptional level. CAR protein from liver nuclear extraction was decreased with chronic alcohol use while CAR protein from total liver changed little. Conclusion Our findings indicated that chronic alcohol uptake selectively regulated bilirubin clearance pathway, accompanied by impairment of translocation of CAR.