法舒地尔通过Rho/ROCK信号通路抑制肝星状细胞黏附、迁移和增殖

Fasudil inhibites HSC adhesion,migration and proliferation via Rho/ROCK pathway

目的观察Rho/Rho激酶(ROCK)信号转导通路抑制剂——法舒地尔,对肝星状细胞(HSC)黏附、迁移和增殖的影响。方法将培养的HSC分为以下5组：对照组；法舒地尔12．5μmol/L组；法舒地尔25μmol/L组；法舒地尔50μmol/L组；法舒地尔100μmol/L组。采用甲苯胺蓝染色法测定细胞黏附率,Boyden Chamber小室测定HSC跨膜迁移数量,用四甲基偶氮唑盐法检测细胞增殖,Western blot检测HSC RhoA、p-MLC(Thr18/Ser19)和α-平滑肌肌动蛋白的表达。结果法舒地尔对HSC的黏附、迁移和增殖均具有抑制作用,随着浓度增加,抑制作用加强；法舒地尔抑制HSC的α-平滑肌肌动蛋白表达,并且对Rho/ROCK信号转导通路关键信号分子p-MLC(Thr18/Ser19)的蛋白表达有抑制作用。结论法舒地尔通过抑制Rho/ROCK信号通路对细胞骨架的调节作用抑制HSC黏附、迁移和增殖。

Objective To observe the effects of fasudil, a Rho/ROCK signaling pathways inhibitor, on adhesion, migration and proliferation of hepatic stellate cells （HSCs）. Methods Cultured HSCs were divided into 5 groups. Fasudil was added to 4 groups in a concentration of 12.5, 25, 50, and 100 μ mol/L, respectively. A group without fasudil added served as untreated controls. The adhesive inhibition effect of fasudil on HSCs was examined by toluidine blue colorimetric assay, the inhibition of migration of HSCs was evaluated by a modified Boyden chamber, and cell proliferation was assessed by MTT assay. The protein levels of RhoA, p-MLC （Thr1 8/Ser19） and α -SMA were assayed by Western blot. Results Fasudil inhibited HSC adhesion, proliferation and LPA-induced migration in a concentration-dependent manner; the protein expressions of α-SMA and p-MLC （Thr1 8/Ser19） were significantly decreased in the presence of fasudil. Conclusion Fasudil can inhibit HSC adhesion, migration and proliferation by supressing the cytoskeleton regulation function of Rho/ROCK signaling pathways.