目的观察川西獐牙菜醇提物对肝细胞胆汁酸转运蛋白Mrp4(multidrug resistanceassociated protein 4,Mrp4)、转录因子Nrf2的调控作用。方法将14只SD大鼠分为2组:正常对照组、川西獐牙菜醇提物组,每组7只。分别用生理盐水、川西獐牙菜醇提物处理7 d后,收集组织标本,免疫组化检测Mrp3、Mrp4在肝脏的表达变化,Western blot及RT-PCR检测胆汁酸转运蛋白Mrp3、Mrp4和转录因子Nrf2、核受体Lrh-1在蛋白及mRNA水平的变化。结果在mRNA水平,RT-q PCR结果显示川西獐牙菜醇提物组肝脏胆汁酸转运蛋白Mrp4表达较正常对照组增加2.1倍,同时,Mrp3表达增加1.7,转录因子Nrf2表达增高1.8倍,核受体Lrh-1表达增高1.4倍;在蛋白水平,Western blot结果表明川西獐牙菜醇提物组肝脏胆汁酸转运蛋白Mrp4表达较正常对照组组增加2.5倍,同时,Mrp3表达增高3.0倍,转录因子Nrf2表达增高2.3倍,核受体Lrh-1表达增高1.3倍,而免疫荧光结果则进一步证实了RT-q PCR与Western blot的结果,显示川西獐牙菜醇提物组Mrp3、Mrp4的表达较正常对照组组明显增强。结论川西獐牙菜醇提物能够刺激肝细胞表面胆汁酸转运蛋白Mrp4表达增高,这种调控作用可能与Nrf2相关。
Objective To investigate the effect of alcohol extract of Swertia mussotii Franch on regulating hepatic bile acid transporter Mrp4 and transcriptional factor Nrf2. Methods Total 14 SD rats were randomly divided into 2 groups (n = 7 ) : a control group treated with normal saline, and a treatment group treated with alcohol extract of Swertia mussotii Franch. After treatment for 7 d, liver samples were collected. The expression levels of bile acid transporters Mrp3 and Mrp4, transcriptional factor Nrf2 and nuclear receptor Lrh-1 were measured by immunofluorescence assay, Western blotting and RT-qPCR. Results Compared with the control group, the mRNA expression levels of Mrp4, Mrp3, Nrf2 and Lrh-1 in the treatment group were increased by 2.1 times, 1.7 times, 1.8 times and 1.4 times, respectively, while the protein expression levels of Mrp4, Mrp3, Nrf2 and Lrh-1 were increased by 2.5 times, 3 times, 2.3 times and 1.3 times, respectively. Furthermore, there was significant positive correlation between the Mrp4 mRNA expression and the Nrf2 mRNA expression in the treatment group. Conclusion The alcohol extract of Swertia mussotii Franch promotes hepatic bile acid transporter Mrp4 expression in rats, which may be mediated by transcriptional factor Nrf2.