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目的:探讨FAK抑制剂(PF562,271)对人肝星状细胞(LX-2)Akt/GSK-3β/β-catenin信号通路的影响,为抗肝纤维化寻找新的作用靶点。方法:CCK-8法检测不同浓度范围(0-7μmol·L~(-1))FAK抑制剂(PF562,271)在12h、24h、48h和72h时对LX-2细胞增殖的影响,Real-time PCR分析PF562,271对LX-2细胞中α-SMA、CollagenⅠmRNA表达水平的影响,western blot检测不同浓度PF562,271对LX-2细胞中Akt、p-Akt、GSK-3β、p-GSK-3β、β-catenin蛋白表达的影响,评价PF562,271对LX-2细胞中Akt/GSK-3β/β-catenin信号通路的影响。结果:PF562,271能够抑制LX-2细胞增殖,且该作用呈剂量和时间相关性;qRT-PCR检测到PF562,271能够抑制LX-2细胞中α-SMA、CollagenⅠmRNA表达,与对照组相比,表达量显著降低(P<0.05);western blot结果显示PF562,271对Akt/GSK-3β的磷酸化表达水平显著降低(P<0.05)。结论:FAK抑制剂能够抑制LX-2细胞增殖,促进其凋亡,其作用机理可能是通过抑制Akt/GSK-3β/β-catenin信号通路从而缓解肝纤维化进程。
Objective: To investigate the effect of FAK inhibitor (PF562,271) on Akt / GSK-3β / β-catenin signaling pathway in human hepatic stellate cells (LX-2) and to find new target for anti-hepatic fibrosis. Methods: The effects of FAK inhibitor (PF562, 271) in different concentrations (0-7μmol·L -1) on the proliferation of LX-2 cells were detected by CCK-8 assay. The effects of Real- The effect of different concentrations of PF562,271 on the expression of Akt, p-Akt, GSK-3β, p-GSK-3β in LX-2 cells was detected by time PCR. 3β and β-catenin in human peripheral blood mononuclear cells, and to evaluate the effect of PF562,271 on Akt / GSK-3β / β-catenin signaling pathway in LX-2 cells. Results: PF562,271 could inhibit the proliferation of LX-2 cells in a dose-and time-dependent manner. PF562,271 could inhibit the expression of α-SMA and CollagenⅠmRNA in LX-2 cells by qRT-PCR as compared with the control group (P <0.05). Western blot results showed that the phosphorylation of Akt / GSK-3β in PF562 and 271 cells was significantly decreased (P <0.05). Conclusion: FAK inhibitor can inhibit the proliferation and promote the apoptosis of LX-2 cells. Its mechanism may be through the inhibition of Akt / GSK-3β / β-catenin signaling pathway to alleviate hepatic fibrosis.