TGF-β1表达对成纤维细胞胶原分泌影响的实验研究

SiRNA-induced silencing of transforming growth factor-β1 on the expression of collagen protein in fibroblasts

目的通过siRNA干扰成纤维细胞中TGF-β1(TGF-β1)的表达,探讨其对成纤维细胞合成Ⅰ型胶原蛋白的影响。方法将siRNA转染兔成纤维细胞,通过RT-PCR,检测siRNA对兔TGF-β1表达的干扰效果,并选筛选出有干扰效果的siRNA。把筛选出的有最佳干扰效率的siRNA,重新转染兔成纤细胞,分别取成纤维细胞培养液行ELISA检测,观察细胞培养液中合成Ⅰ型胶原蛋白变化情况。结果和对照组相比,siTGF-β1对成纤维细胞有明显的干扰效果;siTGF-β1转染成纤维细胞后,转染组细胞分泌Ⅰ型胶原浓度曲线明显低于空白细胞组,胶原蛋白合成明显降低。结论 siRNA可通过干扰TGF-β1的表达抑制成纤维细胞I型胶原蛋白的分泌,从而抑制瘢痕生成,为临床上尿道瘢痕的基因治疗提供了理论依据。

Objective To investigate the effect of small interfering RNA （siRNA） expressing plasmids targeting the TGF- β1 （transforming growth factor-β1,TGF -β1） on the fibroblasts synthesis type I collagen. Methods Constructed siRNA expressing plasmids targeting TGF-β1 were transfected into fibroblasts of rabbit. The effective siRNA was screened out with the method of RT-PCR and transfected into fibroblasts of rabbit again. The concentration of fibroblasts synthesis type I collagen in culture medium was measured with the method of ELISA. Results Compared with the control group,siTGF-β1 of the experimental group had significant interference on fibroblasts. The type I collagen concentration curve in transfected cells was significantly lower than that of the control group. Conclusions TGF-β1 siRNA can inhibit the expression of TGF-β1 and reduce the secretion of type I collagen significantly. The down regulation of fibroblasts synthesis type I collagen by constructed siRNA interfering TGF-β1 expression provides a theory basis for genetic therapy for urethral scar.