血管生成素-2通过膜突蛋白介导内皮细胞通透性增高  

Angiopoietin-2 increases endothelial cell permeability through moesin mediation

作　　者：袁双双 余琴 邓鑫 游婧璨 李永杰 陈妮 王立群 YUAN Shuangshuang;YU Qin;DENG Xin;YOU Jingcan;LI Yongjie;CHEN Ni;WANG Liqun(Basic Medicine Research Innovation Center for cardiometabolic diseases,Ministry of Education,Southwest Medical University,Luzhou 646000,China;Department of Cardiovascular Pharmacology,Southwest Medical University,Luzhou 646000,China)

机构地区：[1]西南医科大学代谢性心血管疾病医药基础研究创新中心,泸州646000 [2]西南医科大学心血管药理系,泸州646000

出　　处：《西南医科大学学报》2024年第3期210-214,共5页Journal of Southwest Medical University

基　　金：国家自然科学基金(81970260);四川省自然科学基金项目(2022NSFSC0671)。

摘　　要：目的探讨血管生成素-2(Angiopoietin-2,Ang-2)对内皮细胞通透性的影响及其潜在分子机制。方法内皮细胞培养于Transwell培养皿,给予Ang-2(200 ng/mL)刺激0、12、24和48 h后检测内皮细胞通透性变化;内皮细胞接种到6孔板,Ang-2刺激0、15、30和60 min后检测膜突蛋白(Moesin)的磷酸化;使用moesin siRNA特异性敲低moesin后,给予Ang-2刺激,检测内皮细胞通透性的改变,并进一步利用免疫荧光染色检测内皮细胞F-actin以及血管内皮-钙黏蛋白(Vascular endothelial cadherin,VE-cadherin)的变化。结果Ang-2(200 ng/mL)刺激内皮细胞0、12、24和48 h后,细胞通透性以时间依赖的方式逐渐增高;Ang-2显著增加内皮细胞moesin磷酸化,而使用Ang-2受体Tie-2的中和抗体Anti-Tie-2处理细胞后,显著抑制了Ang-2介导的moesin磷酸化;Moesin的特异性敲除显著抑制了Ang-2介导的内皮细胞通透性增高、F-actin应力纤维的形成以及VE-cadherin的破坏。结论Ang-2通过moesin介导内皮细胞通透性增高。Objective To probe into the functions of angiopoietin-2(Ang-2)in endothelial cell permeability and the potential molecular mechanisms involved.Methods The endothelial cells were plated onto transwell,followed by stimulation with Ang-2(200 ng/mL)and then the endothelial cell permeability were detected.The endothelial cells cultured in 6-well plates were stimulated with Ang-2(200 ng/mL)for 0,15,30 and 60 min.Then the moesin phosphorylation was detected by western blotting.Moreover,the moesin siRNA was used to knock down the expression of moesin and the effects of moesin knockdown on Ang-2-regulated endothelial cell per⁃meability,stress fiber formation and vascular endothelial cadherin(VE-cadherin)were detected.Results Ang-2 significantly in⁃creased endothelial cell permeability in a time-dependent manner.The activation of moesin was remarkably evoked with Ang-2 treat⁃ment for 15 min and peaked in 30 min.However,the anti-Tie-2 antibody significantly prevented Ang-2-induced moesin phosphoryla⁃tion.Furthermore,knockdown of moesin with moesin siRNA significantly prevented Ang-2-induced endothelial cell permeability in⁃crease,stress fiber formation and VE-cadherin disruption.Conclusion Ang-2 increased endothelial cell permeability through moesin mediation.

关 键 词：血管生成素-2 膜突蛋白 内皮细胞通透性 

分 类 号：R363[医药卫生—病理学]