摘要
目的:研究转化生长因子βⅡ型受体小干扰RNA(TGFβRⅡ-siRNA)对肝星状细胞TGFβR/Smad信号通路的影响。方法:构建TGFβRⅡ-siRNA的表达质粒,将TGFβRⅡ-siRNA质粒采用脂质体转染HSC-T6细胞。转染72 h后,采用半定量RT-PCR检测TGFβRⅡ、Samd2、Smad3、Smad7的mRNA的表达变化,放射免疫法测上清液Ⅲ型前胶原、IV型胶原和透明质酸含量变化。结果:与无关siRNA比较,TGFβRⅡ、Smad2和Smad3 mRNA表达差异有显著性(P<0.01),但Smad7表达差异无显著性,上清液Ⅲ型前胶原、IV型胶原和透明质酸含量明显减少(P<0.01)。结论:TGFβRⅡ-siRNA下调TGFβRⅡ、Samd2、Smad3 mRNA表达,减少肝星状细胞胶原合成,但对Smad7 mRNA无明显下调。
Objective：To investigate the effect of transforming growth factorβreceptor-II（TGF βRII） small interfering RNA（siRNA） on the expression of TGFβRII,Smad 2,Smad 3,Smad 7 in hepatic stellate cells（HSC-T6）.Methods：TGFβRII-siRNA plasmid was constructed,the TGFβRII-siRNA plas-mid transfected into HSC-T6 with liposome metafectene 2000 after 72 hours,the expressions of TGF βRII,Smad 2,Smad3、Smad7 mRNA were detected by reverse transcription polymerase chain reaction（RT-PCR）、The contents of hyalur-onicacid,IV collagen and type Ⅲ procollagen in the supernatants was measured by radioimmunoassay.Results：Compared with scrambled control groupt,he expression of TGFβRII,Smad2 and Smad3 mRNA in TGFβRII-siRNA group decreased remarkably（P 〈0.01）,while the expression of Smad7 mRNA remains unchanged.And The contents of hyaluronic acid,IV collagen and type Ⅲ procollagen in the supernatants decreased remarkably（P 〈0.01）.Conclusion：The TGFβRII-siRNA could decrease the expression of TGFβRII,Smad2 and Smad3 mRNA and reduce the collagen synthesis,but have no effect on the expression of Smad7 mRNA in HSCs.
出处
《温州医学院学报》
CAS
2010年第4期389-391,共3页
Journal of Wenzhou Medical College
基金
浙江省自然科学基金资助项目(Y2080599)
温州市科技局科研基金资助项目(Y20070045)
