RNA干扰对人瘢痕疙瘩成纤维细胞CTGF表达和胶原合成的影响

Impact of RNAi on CTGF expression and collagen secretion in HKFs

目的:研究结缔组织生长因(子Connective tissue growth factor,CTGF)siRNA表达载体对瘢痕疙瘩成纤维细(胞Human keloi d fibroblasts,HKFs)中CTGF表达、胶原蛋白分泌的影响。方法:根据RNA干扰(RNA interference,RNAi)设计原则,设计并构建特异性CTGF siRNA表达载体,转染体外培养HKFs。用荧光实时定量PCR分析细胞中CTGF mRNA表达水平,3H-脯氨酸掺入法测定细胞胶原蛋白分泌水平。结果:与质粒组和对照组比较,CTGF siRNA表达载体组的CTGF mRNA表达降低约59.9%(P<0.05),胶原蛋白分泌明显下降(P<0.05)。结论:CTGF siRNA表达载体能明显抑制HKFs中CTGF表达及胶原蛋白分泌水平,有可能成为临床上治疗瘢痕疙瘩的新方法。

Objective To study the effect of the constructed vecrors expressing siRNA of connective tissue growth factor （CTGF） on CTGF mRNA level and collagen secretion in HKFs. Methods Vectors expressing CTGF siRNA were designed, constructed and transfected into HKFs. The real-time fluorescent quantitative PCR（RQ-PCR）was performed to detect CTGFmRNA level. The collagen secretion was assessed by 3H-proline incorporation method. Results The CTGFmRNA level and the collagen secretion of vecrors group were markedly lower than those of plasmids group （both P〈0.05）. Conclusion Vectors expressing CTGF siRNA can inhibit CTGFmRNA level and the collagen secretion in HKFs. This may be a new way to treat keloid.