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目的:探讨青蒿琥酯(Art)对Akt/GSK-3β/β-catenin信号通路的影响。方法:不同浓度(0,12.5,25,50μg·ml~(-1))Art作用于人源肝星状细胞(LX-2),采用CCK-8法检测细胞增殖情况,并确定给药浓度;给予不同浓度Akt抑制药MK-2206 0~8μmol·L~(-1),Western blot法确定其最佳抑制浓度;给予Art、MK-2206、MK-2206+Art,采用Western blot法检测各组Akt、pAkt、GSK-3β、p-GSK-3β、β-catenin蛋白表达情况。结果:CCK-8法检测细胞存活率,当选用25μg·ml~(-1)Art作用于LX-2细胞24h时细胞存活率约80%,Western blot法确定当MK-2206浓度为6μmol·L~(-1)时,可有效抑制p-Akt的表达;Art组(25μg·ml~(-1))、MK-2206组(6μmol·L~(-1))、MK-2206(6μmol·L~(-1))+Art(25μg·ml~(-1))组与对照组相比,Akt、p-Akt、p-GSK-3β、β-catenin蛋白表达均有显著差异(P<0.05),MK-2206(6μmol·L~(-1))+Art(25μg·ml~(-1))组分别与Art(25μg·ml~(-1))组、MK-2206(6μmol·L~(-1))组比较,GSK-3β、Akt蛋白表达无显著差异(P>0.05),p-Akt、p-GSK-3β、β-catenin蛋白表达显著降低(P<0.01)。结论:Art可通过Akt因子对Wnt/β-catenin信号通路中相关因子产生影响,进而抑制细胞增殖,缓解肝纤维发展进程。
Objective: To investigate the effect of artesunate on Akt / GSK-3β / β-catenin signaling pathway. METHODS: Artesin (0,12.5,25,50μg · ml -1) Art was administered to human hepatic stellate cells (LX-2) at different concentrations. The cell proliferation was detected by CCK-8 assay and the drug concentration ; MK-2206 0-8μmol·L -1 Akt inhibitor MK-2206 was given at different concentrations, and the best inhibitory concentration was determined by Western blot. Art, MK-2206 and MK- Group Akt, pAkt, GSK-3β, p-GSK-3β, β-catenin protein expression. Results: Cell viability was detected by CCK-8 assay. Cell viability was about 80% when 25 μg · ml -1 Art was treated with LX-2 cells for 24 h. Western blot showed that when the concentration of MK-2206 was 6 μmol·L (-1), MK-2206 (6μmol·L -1), MK-2206 (6μmol · L -1), Art group (25μg · ml -1) The expression of Akt, p-Akt, p-GSK-3β and β-catenin were significantly different from those in the control group (P < 0.05), MK-2206 (6μmol·L -1) + Art (25μg · ml -1) group and Art (25μg · ml -1) There was no significant difference in the expression of GSK-3β and Akt (P> 0.05), but the expression of p-Akt, p-GSK-3β and β-catenin were significantly lower than those in the control group (P <0.01) Conclusion: Art can affect the related factors of Wnt / β-catenin signaling pathway by Akt factor, then inhibit cell proliferation and alleviate the development of hepatic fibrosis.