Xiang Shulin, Zeng Yi, Xiong Bin, Qin Yueqiu, Huang Xia, Jiang Yujie, Luo Weigui, Sooranna Suren R, Pinhu Liao
The First Clinical Medical College of Jinan University, Guangzhou, 510630 Guangdong Province China.
Department of Intensive Care Unit, the People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021 China.
J Inflamm (Lond). 2016 Jun 10;13:19. doi: 10.1186/s12950-016-0128-1. eCollection 2016.
Endothelin-1 (ET-1) is involved in pulmonary vascular remodeling. The aim of this study was to investigate the biochemical interactions between PPAR-γ, TGF-β1 and ET-1 in vitro.
A549 cells were pre-treated with S2505 (10 μM), S2871 (10 μM) with/without SB203580 (10 μM) for 60 min following 2 h treatment with 10 ng/mL TGF-β1. A549 cells were also transfected with positive or negative PPAR-γ plasmids for comparison. RT-PCR, ELISA, western blotting and confocal laser scanning microscopy (CLSM) were used to measure the relevant expression of mRNA, protein, mediators of pathways and nuclear factor translocation.
SB203580 inhibited TGF-β1 induced ET-1 expression in A549 cells. S2871 decreased PPAR-γ mRNA and increase TGF-β1-induced ET-1 expression. S2871 increased phosphorylation of p38 MAPK and Smad2. Cells transfected with PPAR-γ negative plasmid increased TGF-β1 induced ET-1 expression, and increased the expression of phospho-p38 MAPK and phospho-Smad2. S2505 increased PPAR-γ mRNA expression, suppressed the increased TGF-β1-induced expression of ET-1. S2505 inhibited TGF-β1 induced phosphorylation of p38 MAPK and Smad2, also the nuclear translocation of Smad2. Cells transfected with PPAR-γ positive plasmid reduced TGF-β1-induced ET-1 expression, and inhibited the expression of phospho-p38 MAPK and phospho-Smad2.
TGF-β1 induced release of endothelin-1 is PPAR-γ dependent in cultured A549 cells.
内皮素-1（ET-1）参与肺血管重塑。本研究旨在体外研究过氧化物酶体增殖物激活受体γ（PPAR-γ）、转化生长因子-β1（TGF-β1）和ET-1之间的生化相互作用。
用10 ng/mL TGF-β1处理A549细胞2小时后，分别用S2505（10 μM）、S2871（10 μM）加或不加SB203580（10 μM）预处理60分钟。A549细胞还用PPAR-γ正或负性质粒转染以作比较。采用逆转录聚合酶链反应（RT-PCR）、酶联免疫吸附测定（ELISA）、蛋白质印迹法和共聚焦激光扫描显微镜（CLSM）检测mRNA、蛋白质、信号通路介质和核因子转位的相关表达。
SB203580抑制TGF-β1诱导的A549细胞中ET-1表达。S2871降低PPAR-γ mRNA水平并增加TGF-β1诱导的ET-1表达。S2871增加p38丝裂原活化蛋白激酶（p38 MAPK）和Smad2的磷酸化。用PPAR-γ阴性质粒转染的细胞增加TGF-β1诱导的ET-1表达，并增加磷酸化p38 MAPK和磷酸化Smad2的表达。S2505增加PPAR-γ mRNA表达，抑制TGF-β1诱导的ET-1表达增加。S2505抑制TGF-β1诱导的p38 MAPK和Smad2磷酸化以及Smad2的核转位。用PPAR-γ阳性质粒转染的细胞降低TGF-β1诱导的ET-1表达，并抑制磷酸化p38 MAPK和磷酸化Smad2的表达。
在培养的A549细胞中，TGF-β1诱导的内皮素-1释放是PPAR-γ依赖性的。
