【摘 要】
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本文旨在研究瞬时受体电位香草素受体4(transient receptor potential vanilloid 4,TRPV4)通道对慢性低氧肺高压(chronic hypoxia-induced pulmonary hypertension,CHPH)大鼠肺微血管内皮细胞(pulmonary microvascular endothelial cells,PMVECs)通透性的影响,以阐明CHPH发生时血管内皮功能障碍的可能机制.Sprague-Dawley大鼠慢性低氧处理21天建立CHPH模型,组
【机 构】
：
福建医科大学1基础医学院;心血管病离子通道信号调控福建省高校重点实验室,福州350112;心血管病离子通道信号调控福建省高校重点实验室,福州350112;中国人民解放军联勤保障部队第九○○医院,福州3
论文部分内容阅读
本文旨在研究瞬时受体电位香草素受体4(transient receptor potential vanilloid 4,TRPV4)通道对慢性低氧肺高压(chronic hypoxia-induced pulmonary hypertension,CHPH)大鼠肺微血管内皮细胞(pulmonary microvascular endothelial cells,PMVECs)通透性的影响,以阐明CHPH发生时血管内皮功能障碍的可能机制.Sprague-Dawley大鼠慢性低氧处理21天建立CHPH模型,组织块贴壁法原代培养PMVECs,采用FITC-葡聚糖法检测细胞通透系数,透射电镜观察紧密连接(tightjunction,TJ)结构,实时荧光定量PCR和蛋白印迹法检测TRPV4和TJ相关蛋白Occludin、Claudin-5、ZO-1表达变化,TRPV4特异性激动剂GSK1016790A(GSK,10 nmol/L)和抑制剂HC-067047(HC,1 μmol/L,0.5 μmol/L)干预后观察PMVECs内Ca2+浓度变化及其对PMVECs通透性的影响.结果显示:慢性低氧处理21天后成功建立CHPH大鼠模型,CHPH大鼠肺血管内皮间TJ结构破坏,PMVECs屏障功能降低,细胞间通透性增高,TJ相关蛋白表达显著减少,TRPV4表达增高(P＜0.01).特异性激活TRPV4后CHPH大鼠PMVECs内Ca2+浓度显著增高,TRPV4特异性抑制剂HC对CHPH大鼠细胞Ca2+增高的抑制效应显著高于正常PMVECs,并可以逆转CHPH大鼠PMVECs的通透性增高,增加三种TJ相关蛋白表达(P＜0.01或P＜0.05).上述结果提示,TRPV4通道通过增加胞内Ca2+浓度,引起CHPH大鼠PMVECs上TJ结构破坏、TJ相关蛋白表达下降,诱发内皮功能障碍.
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