摘要
目的 探讨康柏西普(conbercept)对转化生长因子-β(TGF-β2)诱导人晶状体上皮细胞(HLECs)发生上皮间质转化(EMT)2的逆转机制.方法 体外培养HLEC-SRA01/04细胞并分为对照组、TGF-β2组、conbercept组、TGF-β2+conbercept组.采用MTT法、流式细胞术、划痕实验、Transwell检测细胞增殖、凋亡与迁移,Western blotting 和qRT-PCR 检测EMT相关上皮细胞标记物E-Cadherin、α-SMA、snail、细胞外基质和TGF-β/Smad信号通路相关基因的表达.结果 TGF-β2+conbercept组EMT程度明显减轻,细胞间充质及细胞外基质标志物α-SMA、snail、collagen Ⅰ、collagen Ⅳ、FN1的表达量均明显减少,与TGF-β2组的差异有统计学意义(P<0.05).上皮细胞相关标志物E-Cadherin的蛋白及mRNA的表达上调(P<0.05).Transwell结果显示,与TGF-β2组相比,TGF-β2+conbercept组细胞迁移能力减弱(P<0.05).此外,TGF-β2诱导HLEC-SRA01/04细胞发生EMT过程中发生Smad2/3磷酸化水平升高现象可被康柏西普抑制(P<0.01).结论 康柏西普可能通过TGF-β/Smad信号通路抑制HLEC-SRA01/04细胞EMT过程,具有预防及治疗PCO的能力.
Abstract
Objective To investigate the mechanism by which conbercept reverses transforming growth factor-β2(TGF-β2)-induced epithelial-mesenchymal transition(EMT)in human lens epithelial cells(HLECs).Methods Cultured HLEC SRA01/04 cells were treated with TGF-β2,conbercept,or both,and the changes in cell proliferation,apoptosis,and migration were observed using MTT assay,flow cytometry,scratch assay,and Transwell assay.Western blotting and qRT-PCR were used to detect the changes in the expression of EMT-related epithelial cell markers(E-Cadherin,α-SMA,and Snail),extracellular matrix components,and genes related to the TGF-β/Smad signaling pathway.Results Conbercept significantly reduced TGF-β2-induced EMT of SRA01/04 cells,decreased the expression levels of mesenchymal and extracellular matrix markers α-SMA,Snail,collagen I,collagen IV,and FN1,and upregulated the protein and mRNA expressions of E-cadherin(P<0.05).Transwell assay showed significantly lower cell migration ability in TGF-β2+conbercept group than in TGF-β2 group(P<0.05).Conbercept also inhibited the increase in Smad2/3 phosphorylation levels in HLEC-SRA01/04 cells with TGF-β2-induced EMT(P<0.01).Conclusion Conbercept inhibits TGF-β2 induced EMT by downregulating the expression of pSmad2/3 in TGF-β/Smad signaling pathway,indicating a potential therapeutic strategy against visual loss induced by posterior capsule opacification.
基金项目
安徽省高校自然科学重点项目(KJ2021A0341)
国家科技期刊平台
NSTL
万方数据