EW-7197对TGF-β1诱导人Tenon囊成纤维细胞增殖和迁移的影响及机制  

Effects of EW-7197 on the proliferation and migration of TGF-β1 induced human Tenon fibroblasts and its mechanism

作　　者：温佳敏 郑磊 艾伟鹏 Jia-Min Wen;Lei Zheng;Wei-Peng Ai(Department of Pharmacy,Shenzhen University General Hospital,Shenzhen 518071,Guangdong Province,China;Affiliated Shenzhen Eye Hospital of Jinan University,Shenzhen 518040,Guangdong Province,China)

机构地区：[1]深圳大学总医院药学部,中国广东省深圳市518071 [2]暨南大学附属深圳眼科医院,中国广东省深圳市518040

出　　处：《国际眼科杂志》2023年第10期1617-1620,共4页International Eye Science

基　　金：深圳市科创委基础研究项目(面上项目)(No.JCYJ20190808170609490)。

摘　　要：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。AIM:To investigate the effect of ALK5 inhibitor EW-7197 on the proliferation and migration of human Tenon fibroblasts(HTFs)induced by transforming growth factor-β1(TGF-β1)and its mechanism.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 6.0μmol/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.

关 键 词：青光眼 EW-7197 TENON囊成纤维细胞 转化生长因子β1(TGF-β1) 瘢痕化 

分 类 号：R775[医药卫生—眼科]