Wang Li-Le, Zhu Xiao-Li, Han Shu-Hua, Xu Lu
Department of Respiratory Medicine, Zhongda Hospital, School of Medicine, Southeast University, Nanjing 210009, China.
School of Medicine, Southeast University, Nanjing 210009, China.
Evid Based Complement Alternat Med. 2021 Nov 25;2021:1525619. doi: 10.1155/2021/1525619. eCollection 2021.
To investigate the effect of hypoxia on pulmonary artery endothelial cells and the role of NOTCH3 in endothelial-mesenchymal transition (EnMT) and to provide a research model for pulmonary disease and explain the pathogenesis of the pulmonary disease.
Pulmonary artery endothelial cells were divided into two groups and cultured in normoxic and hypoxic environments, respectively. QPCR, western blot, and immunofluorescence were used to detect endothelial cell-specific marker protein and mRNA expression in each group, and the ability of endothelial cells migration was evaluated by scratch and transwell experiment.
The pulmonary artery endothelial cells in the normoxic group presented a typical pebble-like arrangement, and the endothelial cells in hypoxic culture showed a long spindle appearance. Hypoxia induced high expression of NOTCH3, Jagged-1, Hes1, c-Src, and CSL. Immunofluorescence showed that endothelial cells in hypoxic culture began to express the -SMA, and the expression of vWF increased with hypoxia. Cell viability, scratch, and transwell results showed that endothelial cells in the hypoxic group were more capable of viability and migration than those in the normoxic group. The induction of EnMT by hypoxia can be inhibited by using notch3-specific inhibitor DAPT and Jagged-1. This study also found that miR-7-5p can regulate endothelial NOTCH3, indicating that miRNA is also involved in the process of endothelial-mesenchymal transformation.
Hypoxia promotes the transformation of endothelial cells into mesenchymal cells by opening the NOTCH3 pathway, which lays the foundation for disease progression or clinical prognosis, and is of great significance in the treatment of diseases.
探讨缺氧对肺动脉内皮细胞的影响以及NOTCH3在内皮-间质转化（EnMT）中的作用，为肺部疾病提供研究模型并解释肺部疾病的发病机制。
将肺动脉内皮细胞分为两组，分别在常氧和缺氧环境中培养。采用QPCR、蛋白质免疫印迹法和免疫荧光法检测各组内皮细胞特异性标志物蛋白和mRNA表达，并通过划痕实验和Transwell实验评估内皮细胞迁移能力。
常氧组肺动脉内皮细胞呈典型的鹅卵石样排列，缺氧培养的内皮细胞呈长梭形外观。缺氧诱导NOTCH3、Jagged-1、Hes1、c-Src和CSL高表达。免疫荧光显示，缺氧培养的内皮细胞开始表达α-SMA，且vWF表达随缺氧增加。细胞活力、划痕和Transwell结果显示，缺氧组内皮细胞比常氧组更具活力和迁移能力。使用NOTCH3特异性抑制剂DAPT和Jagged-1可抑制缺氧诱导的EnMT。本研究还发现miR-7-5p可调节内皮细胞NOTCH3，表明miRNA也参与内皮-间质转化过程。
缺氧通过开启NOTCH3通路促进内皮细胞向间充质细胞转化，为疾病进展或临床预后奠定基础，对疾病治疗具有重要意义。
