GW501516通过TGFβ-Smad3信号途径促进人脐静脉内皮细胞PAI-1的表达  

GW501516 Promotes PAI-1 Expression in Human Umbilical Vein Endothelial Cells Through TGFβ-Smad3 Signaling Pathway

作　　者：莫中成[1] 李霞[1] 张大棣[1] 朱明燕[1] 张青海[1] 曾颖[1] 伍荣[1] 鲁艳菊 陈意[1] 易光辉[1] 

机构地区：[1]南华大学医学院心血管疾病研究所,动脉硬化学湖南省重点实验室,衡阳421001

出　　处：《生物化学与生物物理进展》2014年第7期674-681,共8页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目(81270360;81100211);湖南省科技计划资助项目(2013SK3114);湖南省重点学科建设项目(南华大学基础医学学科;湘教发[2011]76号)~~

摘　　要：为探讨过氧化物酶体增殖物激活型受体δ(peroxisome proliferator-activated receptor-δ,PPARδ)激动剂GW501516对人脐静脉内皮细胞纤溶酶原激活物抑制剂1(PAI-1)表达的影响及机制,采用siRNA、TGFβ-Smad3信号通路阻滞剂等处理细胞,经实时定量PCR、Western blot方法分别检测细胞中PAI-1及磷酸化Smad3蛋白的表达.结果显示,与对照组比较,GW501516可诱导人脐静脉内皮细胞(HUVEC)中PAI-1表达,且此效应呈浓度和时间依赖性(P<0.05);siRNA沉默PPARδ的表达后,可阻抑GW501516对HUVEC细胞PAI-1表达的促进作用;TGFβ-Smad3信号通路抑制剂SB-431542与SIS3均可降低HUVEC细胞pSmad3蛋白的表达,而细胞PAI-1表达也随之降低.结果提示,GW501516可促进HUVEC细胞PAI-1的表达,其机制可能与TGFβ-Smad3信号通路有关.Increased plasminogen activator inhibitor-1 （PAI-1） level is the risk of thrombotic disease and atherosclerosis （As）. Our research aims to study the effect and mechanism of PPARβ antagonist GW501516 on the expression of PAl-1. Firstly, human umbilical vein endothelial cells （HUVECs） were incubated with DMEM, and then treated with siRNA and TGFβ-Smad3 inhibitor, respectively. The protein and mRNA expression were examined by Western blotting assays and Real-time quantitative PCR, respectively. The result showed that GW501516 induced PAI-1 mRNA and protein expression in HUVECs compared with control groups （P 〈 0.05）. According to PPAR8 gene, the designed PPAR8 siRNA primer silenced the PPAR8 expression and depressed the induction of GW501516 on PAl-1 expression. Then, the HUVECs were treated by SB431542 or SIS3, which is the TGFβ-Smad3 signaling pathway blocker, and found that PAI-1 expression of cells were down-regulated.At last, we found that SB431542, SIS3 and GW501516 inhibited the expressions of pSmad3 protein. These results suggested that TGF beta-Smad3 signaling pathway involved in the regulation of GW501516-induced PAI-1 expression in HUVECs.

关 键 词：GW501516 纤溶酶原激活物抑制剂1 过氧化物酶体增殖物激活型受体δ 转化生长因子β SMAD3 

分 类 号：R363[医药卫生—病理学] R54[医药卫生—基础医学]