阻抑结缔组织生长因子表达对瘢痕疙瘩成纤维细胞的影响  

The effect of connective tissue growth factor RNA interference on keloid-derived fibroblasts

作　　者：夏炜[1] 潘宝华[1] 刘宾[1] 张曦[1] 郑岩[1] 郭树忠[1] 

机构地区：[1]第四军医大学西京医院全军整形外科研究所,西安710032

出　　处：《中华实验外科杂志》2009年第12期1617-1619,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30371469、30801190）;陕西省自然科学基金资助项目（2006C253）

摘　　要：目的观察结缔组织生长因子（CTGF）RNA干扰对瘢痕疙瘩成纤维细胞（KF）增殖以及Ⅰ型胶原合成的影响。方法设计合成3条CTGF小干扰RNA（siRNA）序列并克隆到pRNAT-6．1／Neo载体中；实时定量逆转录-聚合酶链反应（qRT-PCR）检测KF中CTGF和Ⅰ型胶原的表达，计算KF细胞数目。结果合成的3个CTGFsiRNA载体中CTGFsiRNA3的干扰效果最强，将CT—GFmRNA的相对表达量从3．13±0．17降至0．71±0．02，并能将KF中的Ⅰ型胶原的mRNA水平从2．04±0．10降至0．60±0．04（P〈0．01）。KF对照组5d的细胞数目为（10．50±0．25）×10^4，而CT-GF siRNA3组5d的细胞数目为（6．83±0．29）×10^4（P〈0．01）。结论CTGFRNA干扰能抑制KF的CTGF表达，产生抑制Ⅰ型胶原合成和细胞增殖的生物学作用。Objective To investigate the impact of connective tissue growth factor （CTGF） RNA interference on the proliferation and type Ⅰcollagen synthesis of kcloid-derived fibroblasts （KFs）. Methods Three CTGF small interference RNA （siRNA） sequences were cloned into pRNAT-6.1/Neo vector, and transfected into KFs. The expression of CTGF and typeⅠcollagen was detected by using quantitative real-time PCR （ q RT-PCR）. The cell number of KFs was counted. Results All of the three CTGF siRNA vectors showed inhibitory effects on CTGF mRNA expression in KFs. Among them,the CTGF siRNA3 vector was the most potent one,which decreased CTGF relative expression level from 3.13 ±0.17 to 0.71± 0.02 （ P 〈 0.01 ）. Meanwhile, the expression of type I collagen was decreased from 2.04 ± 0. 10 to 0.60 ±0.04 （P 〈 0.01 ）. The cell number of KF on the day 5 was （ 10.50 ± 0.25 ） ×10^4 in the control group, while decreased to （6.83 ±0.29） ×10^4 in the CTGF siRNA3 group （ P 〈 0.01 ）. Conclusion CTGF RNAi inhibits the CTGF expression in KFs, therefore decreases type I collagen expression and cell proliferation of KFs.

关 键 词：瘢痕疙瘩 成纤维细胞 结缔组织生长因子 

分 类 号：R622[医药卫生—整形外科]