Wang You-juan, Huang Yan, Tang Hui, Liu Xiao-jing
Laboratory of Cardiovascular Diseases, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2009 May;40(3):378-81.
To investigate the effects of small interference RNA (siRNA) targeting connective tissue growth factor (CTGF) on collagen synthesis induced by hypoxia in human lung fibroblasts.
CTGF-siRNA expression plasmid was stablely transfected into MRC-5. The transfected cells were cultured under hypoxia condition (37 degrees C, 5% CO2, 1% O2) for 1, 2, 4, 6 and 12 hours respectively. The expression of CTGF and collagen type I was assessed by real time quantitative PCR and Western blot.
Expression of CTGF mRNA was up-regulated as early as 1 hour in control scrambled-siRNA/MRC-5 cells after exposed to hypoxia condition, which remained the elevated level up to 6 hours. Compared with control cells, the expression of CTGF mRNA and protein induced by hypoxia was decreased significantly in CTGF-siRNA/MRC-5 cells. Moreover, the deposition of collagen type I in CTGF-siRNA/MRC-5 cells was also declined.
The siRNA targeting CTGF gene could efficiently inhibit collagen deposition induced by hypoxia in human lung fibroblasts.
研究靶向结缔组织生长因子（CTGF）的小干扰RNA（siRNA）对人肺成纤维细胞缺氧诱导的胶原合成的影响。
将CTGF-siRNA表达质粒稳定转染至MRC-5细胞。将转染后的细胞分别在缺氧条件（37℃，5%二氧化碳，1%氧气）下培养1、2、4、6和12小时。通过实时定量PCR和蛋白质印迹法评估CTGF和I型胶原的表达。
在暴露于缺氧条件后，对照乱序-siRNA/MRC-5细胞中CTGF mRNA的表达早在1小时就上调，并在6小时内保持升高水平。与对照细胞相比，CTGF-siRNA/MRC-5细胞中缺氧诱导的CTGF mRNA和蛋白质的表达显著降低。此外，CTGF-siRNA/MRC-5细胞中I型胶原的沉积也减少。
靶向CTGF基因的siRNA可有效抑制人肺成纤维细胞缺氧诱导的胶原沉积。
