Relationship of Cx43 regulation of vascular permeability to osteopontin-tight junction protein pathway after sepsis in rats
Relationship of Cx43 regulation of vascular permeability to osteopontin-tight junction protein pathway after sepsis in rats
复制标题
脓毒症后Cx43对血管通透性的调节与骨桥蛋白紧密连接蛋白通路的关系
DOI：
10.1152/ajpregu.00443.2016
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发表时间：
2018-01-01
影响因子：
2.8
通讯作者：
Liu, Liangming
中科院分区：
文献类型：
--
作者：
Zhang, Jie;Yang, Guangming;Liu, Liangming
我们之前的研究表明，连接蛋白（Cx）43参与了严重脓毒症中血管通透性的调节。骨桥蛋白（OPN）已被证实参与动脉粥样硬化、炎症的发生以及细胞的黏附和迁移。目前尚不清楚OPN是否参与脓毒症后Cx43对血管通透性的调节，以及它是否与紧密连接蛋白相关。本研究利用盲肠结扎穿孔（CLP）诱导的脓毒症大鼠和脂多糖（LPS）处理的肺静脉血管内皮细胞（VECs），探究了紧密连接蛋白1（ZO - 1）和克劳丁 - 5（claudin - 5）在Cx43调节血管通透性中的作用及其与OPN的关系。结果显示，在CLP大鼠和LPS处理的肺静脉VECs中，肺静脉中ZO - 1和claudin - 5的表达降低。过表达Cx43的慢病毒诱导了ZO - 1和claudin - 5的降解，而Cx43 RNA干扰慢病毒则消除了LPS诱导的ZO - 1和claudin - 5的降解。在CLP大鼠和LPS处理的肺静脉VECs中，肺静脉的血管通透性和OPN的表达显著增加。通过OPN RNA干扰慢病毒沉默OPN可抑制LPS诱导的血管高通透性。过表达Cx43的慢病毒增加了肺静脉VECs中OPN的表达和血管通透性，并下调了ZO - 1和claudin - 5的表达。通过OPN RNA干扰慢病毒沉默OPN可抑制过表达Cx43的慢病毒对肺静脉VECs中ZO - 1和claudin - 5表达下调以及血管高通透性的影响。转染针对β - 连环蛋白和T细胞因子 - 4（Tcf - 4）的特异性双链RNA可消除Cx43过表达诱导的OPN上调。这些结果表明，OPN参与了脓毒症后Cx43对血管通透性的调节。Cx43通过Tcf - 4/β - 连环蛋白转录途径上调OPN；OPN通过下调紧密连接蛋白ZO - 1和claudin - 5的表达来增加血管通透性。
Our previous study demonstrated that connexin (Cx) 43 participated in the regulation of vascular permeability in severe sepsis. Osteopontin (OPN) has been demonstrated to participate in the occurrence of atherosclerosis, inflammation, as well as the adhesion and migration of cells. It is not clear whether OPN is involved in Cx43 regulating vascular permeability after sepsis and if it is related to tight-junction proteins. with the use of cecal ligation and puncture (CLP)-induced septic rats and lipopolysaccharide (LPS)-treated pulmonary vein vascular endothelial cells (VECs), the role of zona occuldens 1 (ZO-1) and claudin-5 in Cx43 regulation of vascular permeability and its relationship to OPN were investigated in the present study. The results showed that the expression of ZO-1 and claudin-5 in pulmonary vein were decreased in CLP rats and LPS-treated pulmonary vein VECs. Cx43-overexpressed lentivirus induced the degradation of ZO-1 and claudin-5, while Cx43 RNAi lentivirus abrogated the degradation of ZO-1 and claudin-5 induced by LPS. The vascular permeability and expression of OPN in pulmonary veins were significantly increased in CLP rats and LPS-treated pulmonary vein VECs. Silencing OPN by OPN RNAi lentivirus inhibited the vascular hyperpermeability induced by LPS. Overexpressed Cx43 lentivirus increased the expression of OPN and vascular permeability and downregulated the expression of ZO-1 and claudin-5 in pulmonary vein VECs. Silencing OPN by OPN RNAi lentivirus inhibited the effects of Cx43-overexpressed lentivirus on downregulation of ZO-1 and claudin-5 and vascular hyperpermeability in pulmonary vein VECs. Transfection of specific double-stranded RNA targeting to beta-catenin and T-cell factor-4 (Tcf-4) abolished the upregulation of OPN induced by Cx43 overexpression. These results suggest that OPN participates in the regulation of vascular permeability by Cx43 after sepsis. Cx43 upregulation of OPN is via the Tcf-4/beta-catenin transcription pathway; OPN increases vascular permeability by downregulating the expression of the tight junction proteins ZO-1 and claudin-5.