AM966, an Antagonist of Lysophosphatidic Acid Receptor 1, Increases Lung Microvascular Endothelial Permeability through Activation of Rho Signaling Pathway and Phosphorylation of VE-Cadherin.
AM966, an Antagonist of Lysophosphatidic Acid Receptor 1, Increases Lung Microvascular Endothelial Permeability through Activation of Rho Signaling Pathway and Phosphorylation of VE-Cadherin.
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DOI：
10.1155/2017/6893560
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发表时间：
2017
影响因子：
4.6
通讯作者：
Zhao J
中科院分区：
文献类型：
--
作者：
Cai J;Wei J;Li S;Suber T;Zhao J
维持肺内皮屏障的完整性对于减轻肺损伤的严重程度是重要的。溶血磷脂酸（LPA）调节细胞运动、细胞骨架重排和细胞生长。LPA受体1（LPA 1）的敲低已显示减轻肺损伤和肺纤维化。AM 966是一种具有抗纤维化作用的LPA 1拮抗剂，被认为是未来的抗纤维化药物。在这里，我们报告了AM 966的一个意想不到的效果，它增加了肺内皮屏障的通透性。采用细胞-基质电传感（ECIS）系统测定人肺微血管内皮细胞（HLMVECs）的通透性。AM 966以剂量依赖性方式立即降低跨内皮电阻（TEER）值。VE-钙粘蛋白和f-肌动蛋白双重免疫染色显示，AM 966增加内皮细胞之间的应力纤维和间隙形成。AM 966通过激活RhoA/Rho激酶途径诱导肌球蛋白轻链（MLC）磷酸化。与LPA处理不同，AM 966对细胞外信号调节激酶（Erk）的磷酸化没有影响。此外，在LPA 1沉默细胞中，我们观察到AM 966增加的肺内皮通透性以及VE-钙粘蛋白和粘着斑激酶（FAK）的磷酸化减弱。这项研究表明，AM 966诱导肺内皮屏障功能障碍，这是由LPA 1介导的RhoA/MLC激活和VE-钙粘蛋白磷酸化调节的。
Maintenance of pulmonary endothelial barrier integrity is important for reducing severity of lung injury. Lysophosphatidic acid (LPA) regulates cell motility, cytoskeletal rearrangement, and cell growth. Knockdown of LPA receptor 1 (LPA1) has been shown to mitigate lung injury and pulmonary fibrosis. AM966, an LPA1 antagonist exhibiting an antifibrotic property, has been considered to be a future antifibrotic medicine. Here, we report an unexpected effect of AM966, which increases lung endothelial barrier permeability. An electric cell-substrate sensing (ECIS) system was used to measure permeability in human lung microvascular endothelial cells (HLMVECs). AM966 decreased the transendothelial electrical resistance (TEER) value immediately in a dose-dependent manner. VE-cadherin and f-actin double immunostaining reveals that AM966 increases stress fibers and gap formation between endothelial cells. AM966 induced phosphorylation of myosin light chain (MLC) through activation of RhoA/Rho kinase pathway. Unlike LPA treatment, AM966 had no effect on phosphorylation of extracellular signal-regulated kinases (Erk). Further, in LPA1 silencing cells, we observed that AM966-increased lung endothelial permeability as well as phosphorylation of VE-cadherin and focal adhesion kinase (FAK) were attenuated. This study reveals that AM966 induces lung endothelial barrier dysfunction, which is regulated by LPA1-mediated activation of RhoA/MLC and phosphorylation of VE-cadherin.