机械牵张对人肺血管内皮细胞通透性的影响及其分子机制  

Effect of mechanical stretch on permeability of human pulmonary vascular endothelial cells and underlying molecular mechanism

作　　者：王亚 兰媛 傅威 刘晓青[1] 黄勇波[1] 毛璞[2] 黎毅敏[1] Wang Ya;Lan Yuan;Fu Wei;Liu Xiaoqing;Huang Yongbo;Mao Pu;Li Yimin(The First Affiliated Hospital of Guangzhou Medical University,Guangzhou Institute of Respiratory Disease,State Key Laboratory of Respiratory Diseases,Guangzhou 510120,China;Department of Infection Management,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,China)

机构地区：[1]广州医科大学第一附属医院,广州呼吸健康研究院,呼吸疾病国家重点实验室,510120 [2]广州医学院第一附属医院医院感染管理科,510120

出　　处：《中华重症医学电子杂志》2019年第2期151-158,共8页Chinese Journal Of Critical Care & Intensive Care Medicine(Electronic Edition)

基　　金：国家自然科学基金(81770079);广东省自然科学基金(2017A030313781和2017A030313712);广东省科技计划(2016A020215170)

摘　　要：目的观察机械牵张对人肺动脉内皮细胞(HPAEC)和人肺微血管内皮细胞(HPMVEC)通透性的影响及对通透性关键蛋白VE-cadherin、Claudin-5和Caveolin-1表达的调控。方法采用机械牵张装置Flexcell FX-5000T对HPAEC(ATCC)和HPMVEC(ATCC)施加0.5 Hz频率10%或20%牵张应力。应用qRT-PCR及Western Blot方法检测机械牵张前后内皮通透性关键蛋白VEcadherin、Claudin-5、Caveolin-1 mRNA和蛋白含量表达变化。采用细胞动态分析仪以及Transwell小室/异硫氰酸荧光素(FITC)-白蛋白法检测机械牵张后的HPAEC细胞和HPMVEC细胞通透性改变。结果20%机械牵张后,与对照组比较,HPAEC细胞通透性关键蛋白VE-cadherin mRNA和蛋白表达、Claudin-5 mRNA和蛋白表达和Caveolin-1 mRNA表达均下降(P<0.05),而Caveolin-1蛋白水平较对照组无明显变化;HPMVEC细胞通透性关键蛋白VE-cadherin mRNA和蛋白表达,Caveolin-1 mRNA和蛋白表达均下降(P<0.05),而Claudin-5 mRNA和蛋白水平较对照组无明显变化。结论机械牵张会导致肺血管内皮通透性增高,是通过下调通透性关键蛋白的表达,并且两种细胞的通透性影响机制有差异,HPAEC牵涉紧密连接,HPMVEC牵涉跨内皮细胞途径。Objective To observe the effect of mechanical stretch on the permeability of human pulmonary artery endothelial cells(HPAECs)and human pulmonary microvascular endothelial cells(HPMVECs)and to examine whether the expression of the key permeability-related proteins VE-cadherin,Claudin-5,and Caveolin-1 is involved in this process.Methods The mechanical stretch device Flexcell FX-5000T was used to apply 10%or 20%tensile stress to HPAECs and HPMVECs at a frequency of 0.5 Hz.qRT-PCR and Western blot were used to detect the mRNA and protein expression of VE-cadherin,Claudin-5,and Caveolin-1 in cells before and after mechanical stretch.Mechanical stretched HPAEC cells and HPMVEC cells were monitored by a cell dynamic analyzer and Transwell chamber/FITC-albumin assay to detect their permeability.Results In HPAECs after mechanical stretch,the expression of VE-cadherin mRNA and protein,Claudin-5 mRNA and protein,and Caveolin-1 mRNA decreased significantly compared with control cells(P<0.05),while the level of Caveolin-1 protein did not change significantly compared with the control group.In HPMVECs after mechanical stretch,the expression of VE-cadherin mRNA and protein and Caveolin-1 mRNA and protein decreased significantly compared with control cells(P<0.05),while the levels of Claudin-5 mRNA and protein did not change significantly compared with the control group.Conclusion Mechanical stretch leads to increased permeability of pulmonary vascular endothelial cells,which is caused by down-regulation of the expression of permeability key proteins,but the permeability mechanisms of the two cell types are different:HPAECs involve tight junctions and HPMVECs involve the transendothelial pathway.

关 键 词：机械牵张 人肺动脉内皮细胞 细胞通透性 内皮通透性关键蛋白VEcadherin、Claudin-5、Caveolin-1 

分 类 号：R563.8[医药卫生—呼吸系统]