目的：探讨ALK5抑制剂EW-7197对转化生长因子β1(TGF-β1)诱导的人Tenon囊成纤维细胞(HTFs)增殖和迁移的影响及其机制。

方法：采用MTS法检测细胞增殖率，并摸索EW-7197最佳的作用浓度和作用时间。之后将细胞分为3个组，分别为正常对照组、TGF-β1诱导组和TGF-β1+EW-7197处理组，采用Transwell小室观察各组细胞迁移情况，采用Western blot检测各组细胞Fibronectin、α-SMA、p-Smad2、p-Smad3的蛋白表达水平。

结果：MTS结果显示，不同处理条件的EW-7197以6.0μmol/L作用24h的细胞增殖率最低(均P<0.01)。Transwell小室实验结果显示，TGF-β1诱导组的细胞迁移数量为228.0±17.0个/视野，明显高于正常对照组(149.0±15.0个/视野)和TGF-β1+EW-7197处理组(46.0±8.0个/视野)(均P<0.01)。Western blot检测结果显示，TGF-β1诱导组细胞中Fibronectin、α-SMA、p-Smad2、p-Smad3相对表达量高于正常对照组和TGF-β1+EW-7197处理组(均P<0.001)。

结论：EW-7197能够通过TGF-β/Smad信号通路抑制TGF-β1诱导的HTFs增殖和迁移。

METHODS: The cell proliferation rate was detected by MTS assay, and the optimal concentration and time of EW-7197 were explored. Then HTFs were divided into three groups: normal control group, TGF-β1 induced group and TGF-β1+EW-7197 group. Cell migration was observed by Transwell assay. The protein expression levels of Fibronectin, α-SMA, as well as the phosphorylated Smad2, Smad3(p-Smad2, p-Smad3)were measured by Western blot.

RESULTS: MTS assay showed that the proliferation rate of cells treated with 6.0 μmol/L EW-7197 for 24h was the lowest(all P<0.01). Transwell assay showed that the migrated number of HTFs in TGF-β1 induced group was 228.0±17.0/field, which was significantly more than that in normal control group(149.0±15.0/field)and TGF-β1+EW-7197 group(46.0±8.0/field; all P<0.01). Western blot showed that the protein relative expression levels of Fibronectin, α-SMA and p-Smad2, p-Smad3 of HTFs in TGF-β1 induced group were significantly higher than that in normal control group and TGF-β1+EW-7197 group(all P<0.001).

CONCLUSION:EW-7197 can suppress the proliferation and migration of TGF-β1-induced HTFs through TGF-β/Smad signaling pathways.