目的：探讨人转化生长因子TGF-β²特异性siRNA真核表达载体转染人结膜囊成纤维细胞后对其TGF-β² mRNA表达的影响。 方法：体外分离并培养人结膜囊成纤维细胞，在培养后传代3次的细胞以TGF-β²特异性siRNA真核表达载体进行转染，并以未转染细胞作为对照。转染后分别于24，48和72h收集细胞，采用RT-PCR技术检测TGF-β²特异性siRNA真核表达载体对TGF-β² mRNA表达的影响。 结果：分离的人结膜囊成纤维细胞于接种约4h左右开始贴壁，同时细胞变长成为梭形，表现出明显的成纤维细胞特性，约36h后达到融合状态；RT-PCR结果示：与对照组相比，转染24,48和72h后的细胞TGF-β2表达抑制率分别为17.40%,52.80%和79.20%,抑制效率呈现随时间延长有所加强的趋势。 结论：TGF-β²特异性siRNA真核表达载体能抑制人结膜囊成纤维细胞TGF-β²mRNA的表达。

AIM: To investigate the effects of siRNA for transforming growth factor β2(TGF-β²) on the expression of TGF-β² mRNA in human Tenon-s capsule fibroblasts. METHODS: Human Tenon- s fibroblasts were transferred by TGF-β² siRNAs vector after separated, cultured and passaged three in vitro. RT-PCR was performed to evaluate the levels of TGF-β² mRNA at 24, 48 and 72 hours after transferred in transferred cells, and untransferred cells were as the controls. RESULTS: Separated human Tenon-s fibroblasts attached in 4 hours and confluent monolayer cells formed in 36 hours. The adherent cells displayed fibroblast-like or spindle shape by microscope observation. The expression of TGF-β2 mRNA of transferred group was reduced significantly in comparison with the control group，expression inhibition rates were 17.40%, 52.80% and 79.20% respectively in 24, 48 and 72 hours after transferred,and this interference function was strengthened with the time extending. CONCLUSION:The vector of siRNA specific for TGF-β² is of potent interference ability for TGF-β² mRNA expression in human Tenon-s capsule fibroblasts.

深圳市科技计划资助项目(No.2009036)