摘要
目的探讨JAK/STAT通路在IL-4对肝星状细胞(HSC)中Ⅰ型胶原基因表达的影响及其作用机制。方法用RT-PCR法和ELISA法分别检测不同浓度IL-4对人肝星状细胞系LX-2中Ⅰ型胶原mRNA表达和蛋白合成的影响;用Western blot法和RT-PCR法分别观察JAK1抑制剂AG490对IL-4诱导的JAK1磷酸化以及Ⅰ型胶原mRNA表达的影响;用Western blot法和RT-PCR法分别观察LX-2转染STAT6-ASON对IL-4诱导的STAT6磷酸化以及Ⅰ型胶原mRNA表达的影响。结果 IL-4诱导LX-2中Ⅰ型胶原mRNA表达及其蛋白的合成,呈剂量依赖性效应,且IL-4浓度为50μg/L时,胶原mRNA和蛋白水平均开始出现显著差异(P<0.001);与空白对照组相比,JAK1和STAT6磷酸化水平分别约为4.8倍和4.0倍,Ⅰ型胶原mRNA表达则分别为4.0倍和5.2倍;而AG490和LX-2转染STAT6-ASON则可以分别阻断IL-4诱导的JAK1磷酸化和STAT6磷酸化以及相应的Ⅰ型胶原mRNA的表达。结论 JAK/STAT信号传导通路参与调节IL-4诱导HSC中Ⅰ型胶原基因表达,并在肝纤维化发生过程中发挥重要的作用。
Objective To investigate the effects of IL-4 on type Ⅰ collagen mRNA expression and protein production,and the roles of Janus kinase/signal transducers and activators transcription（JAK/STAT） signaling transduction pathway in increased type Ⅰ collagen mRNA expression stimulated by IL-4 in activated hepatic stellate cells（HSCs）.Methods First,type Ⅰ collagen mRNA expression and protein production induced by IL-4 at different doses in a human HSC cell line,LX-2 was determined by RT-PCR and ELISA.Second,the effects of JAK inhibitor AG490 on JAK1 phosphorylation and type Ⅰ collagen mRNA expression stimulated by IL-4 were detected by Western blot and RT-PCR.Third,the roles of transfection with STAT6 antisense oligonucleotide（STAT6-ASON） in STAT6 phosphorylation after IL-4 were detected by Western blot.Finally,the effect of transfection with STAT6-ASON on type Ⅰ collagen mRNA expression after IL-4 was measured by RT-PCR.Results IL-4 increased type Ⅰ collagen mRNA expression and protein production in a dose-dependent manner in LX-2.When IL-4 was induced at the concentration 50 μg/L,the mRNA and protein expressions of type Ⅰ collagen were significantly increased（P0.001）.Compared with the control group,the levels of JAK1 and STAT6 phosphorylation were approximately 4.8 fold and 4.0 fold,and the expressions of their type Ⅰ collagen were 4.0 fold and 5.2 fold respectively;AG490 and STAT6-ASON could block phosphorylation levels of JAK1 and STAT6 and their type Ⅰ collagen mRNA expression.Conclusions JAK/STAT signaling pathway is involved in regulating IL-4-induced type Ⅰ collagen mRNA expression and protein production in activated HSC.
出处
《基础医学与临床》
CSCD
北大核心
2012年第4期423-427,共5页
Basic and Clinical Medicine
基金
安徽省高校优秀青年人才基金项目(2010SQRL065)
