转化生长因子-β_1抗体对人翼状胬肉成纤维细胞增殖及TGF-β_1-Smad4信号转导通路的影响

Effects of TGF-β_1 antibody on human pterygium fibroblasts proliferation and TGF-β_1-Smad4 signaling pathway

目的探讨转化生长因子-β1（TGF-β1）抗体对体外培养的翼状胬肉成纤维细胞增殖的抑制作用及对TGF-β1-Smad4信号转导通路的影响,评估其在翼状胬肉发病机制中的作用。方法人翼状胬肉切除后,体外培养其成纤维细胞并传至4~6代时,分别用6.25、12.5、25、50mg/mL TGF-β1抗体处理24、48、72h,采用MTT法检测成纤维细胞的生长抑制率,RT-PCR检测TGF-β1mRNA表达,Western blot检测Smad4蛋白的表达。所得数据采用SPSS19.0统计软件处理。结果 TGF-β1抗体可不同程度抑制成纤维细胞的增殖;TGF-β1抗体处理组TGF-β1mRNA的表达明显低于对照组,且不同浓度组间差异有统计学意义（P〈0.05）,随着浓度增大和作用时间延长,TGF-β1抗体处理组TGF-β1 mRNA的表达量呈逐渐降低趋势。不同浓度的TGF-β1抗体对Smad4的蛋白表达具有明显增强作用,且各组间差异有统计学意义（P〈0.05）。结论 TGF-β1抗体对人翼状胬肉体外培养的成纤维细胞生长具有明显抑制作用,其可能通过调节TGF-β1和Smad4的表达而实现。

Objective To investigate the effects of transforming growth factor--β1 （TGF-β1） antibody on pterygium fibroblasts proliferation in vitro and TGF-β1-Smad4 signaling pathway to assess the related mechanisms. Methods Pterygium was cultured in vitro after cut from the eyes, and the fibroblasts were subculturcd continuously for 4-6 passages, which were then handled by different concentrations of TGF-β1 antibody for 24, 48 and 72 h, respectively. The inhibition rate of TGF-β1 antibody on pterygium fibroblasts was observed with MTT, and the expression of TGF-β1 mRNA was observed with RT-PCR. Western blot was used to detect the expression of Smad4 protein. The data were processed with SPSS 19.0 softwarc. Results Different concentrations of TGF-~3~ antibody had a different inhibitory effect on the proliferation of pterygium fibroblasts. The expression of TGF-β1 mRNA was significantly lower in treatment groups than that in control group, and it diffcred significantly found among all the treatment groups （P〈0.05）. The higher the dose and the longer the time, the lower expression of TGF-β1, mRNA became. Different dose of the antibody could increase the expression of Smad4 protein, and there were significant differences among the treatment groups （P 〈 0. 05）. Conclusion TGF-β1 antibody has significantly inhibitory cffects on the proliferation of pterygium fibroblasts in vitro. The effects may be partially related to regulating the expressions of TGF-β1 and Smad4.