Wang Weisen, Wang Zhi, Tian Dingyuan, Zeng Xi, Liu Yangdong, Fu Qining, Liang Anlin, Zhang Yi, Gao Qiangguo, Cheng Jizhong, Wang Yun
Department of Cell Biology, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, China.
Cadet Battalion, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing, China.
Cell Physiol Biochem. 2018;49(3):985. doi: 10.1159/000493229. Epub 2018 Sep 7.
BACKGROUND/AIMS: Neointimal hyperplasia is responsible for stenosis, which requires corrective vascular surgery, and is also a major morphological feature of many cardiovascular diseases. This hyperplasia involves the endothelial-to-mesenchymal transition (EndMT). We investigated whether integrin β3 can modulate the EndMT, as well as its underlying mechanism.
Integrin β3 was overexpressed or knocked down in human umbilical vein endothelial cells (HUVECs). The expression of endothelial markers and mesenchymal markers was determined by real-time reverse transcription PCR (RT-PCR), immunofluorescence staining, and western blot analysis. Notch signaling pathway components were detected by real-time RT-PCR and western blot analysis. Cell mobility was evaluated by wound-healing, Transwell, and spreading assays. Fibroblast-specific protein 1 (FSP-1) promoter activity was determined by luciferase assay.
Transforming growth factor (TGF)-β1 treatment or integrin β3 overexpression significantly promoted the EndMT by downregulating VE-cadherin and CD31 and upregulating smooth muscle actin α and FSP-1 in HUVECs, and by enhancing cell migration. Knockdown of integrin β3 reversed these effects. Notch signaling was activated after TGF-β1 treatment of HUVECs. Knockdown of integrin β3 suppressed TGF-β1-induced Notch activation and expression of the Notch downstream target FSP-1.
Integrin β3 may promote the EndMT in HUVECs through activation of the Notch signaling pathway.
背景/目的：内膜增生会导致血管狭窄，这需要进行血管矫正手术，并且内膜增生也是许多心血管疾病的主要形态学特征。这种增生涉及内皮-间充质转化（EndMT）。我们研究了整合素β3是否能调节EndMT及其潜在机制。
在人脐静脉内皮细胞（HUVECs）中过表达或敲低整合素β3。通过实时逆转录PCR（RT-PCR）、免疫荧光染色和蛋白质印迹分析来测定内皮标志物和间充质标志物的表达。通过实时RT-PCR和蛋白质印迹分析检测Notch信号通路成分。通过伤口愈合实验、Transwell实验和铺展实验评估细胞迁移能力。通过荧光素酶实验测定成纤维细胞特异性蛋白1（FSP-1）启动子活性。
转化生长因子（TGF）-β1处理或整合素β3过表达通过下调HUVECs中的血管内皮钙黏蛋白（VE-cadherin）和CD31以及上调平滑肌肌动蛋白α和FSP-1，并增强细胞迁移，显著促进了EndMT。敲低整合素β3可逆转这些作用。TGF-β1处理HUVECs后Notch信号被激活。敲低整合素β3可抑制TGF-β1诱导的Notch激活以及Notch下游靶点FSP-1的表达。
整合素β3可能通过激活Notch信号通路促进HUVECs中的EndMT。
