青蒿琥酯抑制肝星状细胞microRNA-154/β-catenin治疗肝纤维化的机制研究

Mechanism research on Artesunate in the treatment of liver fibrosis by inhibiting miR-154/β-catenin in hepatic stellate cell

目的探讨青蒿琥酯抑制肝星状细胞microRNA-154/β-catenin治疗肝纤维化的机制。方法培养大鼠肝星状细胞,用不同浓度(0、5、10、20、30、40μg/mL)青蒿琥酯作用于大鼠肝星状细胞,采用免疫印迹法(Western blot)检测青蒿琥酯作用后细胞内β-eatenin蛋白的表达;实时定量RT-PCR检测青蒿琥酯作用后细胞内的miR-154和β-eatenin mRNA的相对表达。结果 0-40μg/mL青蒿琥酯作用细胞24 h后,免疫印迹法结果显示,β-catenin蛋白量逐渐降低,并呈剂量相关性(P<0.05);实时荧光定量PCR结果显示青蒿琥酯作用后,β-catenin mRNA表达量及miR-154表达量逐渐降低,并呈剂量相关性(P<0.05)。结论青蒿琥酯可能是通过抑制microRNA-154表达作用于Wnt/β-catenin信号通路及来抑制肝纤维化的发生,microRNA-154与Wnt/β-catenin信号通路可能存在一定的相关性。

Objective To investigate the mechanism of Artesunate in the treatment of liver fibrosis by inhibiting miR-154/β-catenin in hepatic stellate cell.Methods HSC-T6 cells were cultured in vitro with different concentrations（0,5,10,20,30,40 μg/mL） of Artesunate on rat hepatic stellate cells.Western blot was used to detect the expression ofβ-catenin protein after drug administration.RT-PCR was used to detect the expression of β-catenin mRNA and microRNA-154 after drug administration.Results After treatment with artesunate for 24 h,the results of Western blot showed that the expression of β-catenin protein significantly decreased in a dose-dependent manner（P〈0.05）.The results of RT-PCR showed that the expression of β-catenin mRNA and microRNA significantly decreased in a dosedependent manner（P〈0.05）.Conclusion Artesunate may suppress the occurrence of liver fibrosis through inhibiting the expression of microRNA-154 on Wnt/β-catenin signaling pathway.There may exist a certain correlation between miR-154 and the Wnt/β-catenin signaling pathway.