摘要
【目的】观察表达转化生长因子(TGF)小干扰RNA(siRNAs)的siTGF质粒,对肝星状细胞激活信号的影响。【方法】通过构建表达TGF之siRNAs的siTGF质粒,将6μg siTGF转染HSC-T6细胞质粒后,分别在d1,d3,d5提取细胞总mRNA,并收集细胞上清液,采用RT-PCR检测其Smad3、Smad7的mRNA的表达变化,放射免疫法测上清液Ⅲ型前胶原含量变化。【结果】smad3s、mad7的mRNA在d1,d3,d5分别下调为0%、61%、70%和0%、15%、16%,上清液Ⅲ型前胶原含量为0%、(42±5)%(、55±7)%。【结论】TGF-β1siRNA下调肝星状细胞激活信号,减少胶原合成,表明TGF-β1siRNA有望成为抗肝纤维化新的手段。
[Objective]To observe the effect of siTGF plasmid expressing small interfering RNAs（siRNA） that target TGF-β1 on collagen and activation signing in hepatic stellate cells （HSC）. [Methods]siTGF plasmid, which could transcribe TGF-β1 small interference RNA was constructed, 6μg siTGFβ1, transfected into HSC-T6 with liposome metafectene at 1,3,5 day, The expressions of Smad3 ,Smad7 mRNA were detected by reverse transcriptionpolymerase chain reaction （RT-PCR） ,Contents of type Ⅲ pro-collagen in the supernatants were determined by radioimmunoassay. [Results]The expression of Smad3 and Smad7 at mRNA levels were decreased by 0 % ,61%, 70% and 0 %, 15%, 16% respectively. The contents of type Ⅲ pro-collagen in the supernatants decreased by 0 %, 42 %± 5 %, 55 % ±7% respectively[Conclusion] TGF-β1 siRNA can diminish TGFβ1, signaling underlying HSC activation and decrease collagen synthesis ,it would probably provide a new therapeutic approach for liver fibrosis.
出处
《医学临床研究》
CAS
2006年第6期875-878,共4页
Journal of Clinical Research
