摘要
目的研究小干扰RNA（siRNA）沉默结缔组织生长因子（CTGF）对大鼠转化生长因子（TGF）β／Smads信号通路的影响。方法将化学合成CTGF siRNA转染肝星状细胞（HSC）T6和经门静脉注入CCl4诱导6周的肝纤维化大鼠，设空白及随机siRNA对照，抽提HSCT6及大鼠肝组织总RNA和蛋白质，应用Western blot和RT-PCR检测HSCT6及肝组织CTGF及TGF βl、Smad2、3、7蛋白质和基因表达。结果与空白对照组相比，siRNA能显著下调HSC T6 CTGF蛋白表达，以48h最明显，CTGF蛋白表达下调94％±4％（t=46．196，P〈0．01），而TGF β1、Smad2、3、7mRNA表达差异无统计学意义；模型组及对照siRNA组，CCla诱导的大鼠肝组织CTGF和TGFp1蛋白表达明显上调，与模型组相比，CTGF siRNA组大鼠肝组织CTGF及TGF β1蛋白表达分别下调95％±2％（，=21．234，P〈0．01）和74％±8％（F=13．464，P〈0．05），但Smad2和Smad7蛋白表达无明显改变。结论沉默CTGF基因表达对大鼠肝TGF β／Smads信号具有阻抑作用。
Objective To investigate the effects of small interfering RNA targeting connective tissue growth factor （CTGF） on rat transforming growth factor beta （TGF beta）/Smads signal pathway. Methods Chemically synthetic siRNA targeting CTGF was transfected into HSC T6 and then they were injected into rat livers through their intraportal veins. At the same time these rats also received CCl4 subcutaneously every three days for 6 consecutive weeks. Untreated HSC T6 or/and rats with random siRNA treatment served as controls. Total RNA or/and protein in HSC T6 and rat hepatic tissues were extracted. The expressions of CTGF and TGF beta 1, Smad2, 3 and 7 genes were detected by reverse transcription-polymerase chain reaction （RT-PCR） and/or Western blot. Results CTGF siRNA significantly reduced the expression of CTGF protein in HSC T6. At 48 h after CTGF siRNA treatment, the down-regulation of CTGF protein was the most significant, up to 94%±4% （t = 46.196, P 〈 0.01）, but the expressions of TGF beta 1, Smad2, 3 and 7 mRNA showed no differences in HSC T6 compared with the blank controls. Six weeks after CCh injections, prominent up-regulations were observed in the gene expressions of CTGF and TGF beta 1 in saline control or siRNA-treated rat livers. Administering CTGF siRNA for six weeks markedly attenuated the induction of CTGF and TGF beta 1 genes; the expressions of CTGF and TGF beta I protein decreased by 95%±2% （F = 21.234, P 〈 0.01） and 74%±8% （F = 13.464, P 〈 0.05）, respectively, whereas Smad2, 7 protein expressions were not affected. Conclusion Silencing the CTGF gene can suppress the TGF beta/Smads signal pathway in rat livers.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2008年第11期840-843,共4页
Chinese Journal of Hepatology
关键词
肝纤维化
结缔组织生长因子
小干扰RNA
转化生长因子Β
Liver fibrosis
Connective tissue growth factor
Small interfering RNA
Transforming growth factor beta
