Intermittent hypoxia-induced endothelial barrier dysfunction requires ROS-dependent MAP kinase activation
Intermittent hypoxia-induced endothelial barrier dysfunction requires ROS-dependent MAP kinase activation
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DOI：
10.1152/ajpcell.00313.2013
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发表时间：
2014-04-01
影响因子：
5.5
通讯作者：
Prabhakar, Nanduri R.
中科院分区：
文献类型：
--
作者：
Makarenko, Vladislav V.;Usatyuk, Peter V.;Prabhakar, Nanduri R.
本研究的目的是确定模拟呼吸暂停伴间歇性缺氧 (IH) 对内皮屏障功能的影响并评估潜在机制。实验对暴露于 IH 的人肺微血管内皮细胞进行了实验，该循环包括 1.5% O-2 30 秒，然后 20% O2 5 分钟。 IH 降低跨内皮电阻 (TEER)，表明内皮屏障功能减弱。 IH 对 TEER 的影响是刺激依赖性的，并且在复氧后是可逆的。 IH 暴露的细胞表现出应力纤维形成和皮质素、血管内皮钙粘蛋白和 zona occlusionns-1 连接蛋白的重新分布，以及细胞-细胞边界处细胞间隙的增加。细胞外信号调节激酶 (ERK) 和 c-jun NH2 末端激酶 (JNK) 在 IH 暴露的细胞中被磷酸化。抑制 ERK 或 JNK 可以阻止 IH 诱导的 TEER 降低以及细胞骨架和连接蛋白的重组。 IH 增加活性氧 (ROS) 水平，而五氯化四 (1-甲基-4-吡啶基) 锰 (III) 卟啉（一种膜渗透性抗氧化剂）可阻止 ERK 和 JNK 磷酸化以及 IH 诱导的内皮屏障功能变化。这些结果表明，IH 通过 ROS 依赖性 MAP 激酶激活导致细胞骨架和连接蛋白重组，从而导致内皮屏障功能障碍。
The objective of the present study was to determine the impact of simulated apnea with intermittent hypoxia (IH) on endothelial barrier function and assess the underlying mechanism(s). Experiments were performed on human lung microvascular endothelial cells exposed to IH-consisting alternating cycles of 1.5% O-2 for 30s followed by 20% O2 for 5 min. IH decreased transendothelial electrical resistance (TEER) suggesting attenuated endothelial barrier function. The effect of IH on TEER was stimulus dependent and reversible after reoxygenation. IH-exposed cells exhibited stress fiber formation and redistribution of cortactin, vascular endothelial-cadherins, and zona occludens-1 junction proteins along with increased intercellular gaps at cell-cell boundaries. Extracellular signal-regulated kinase (ERK) and c-jun NH2-terminal kinase (JNK) were phosphorylated in IH-exposed cells. Inhibiting either ERK or JNK prevented the IH-induced decrease in TEER and the reorganization of the cytoskeleton and junction proteins. IH increased reactive oxygen species (ROS) levels, and manganese (III) tetrakis (1-methyl-4-pyridyl) porphyrin pentachloride, a membrane-permeable antioxidant, prevented ERK and JNK phosphorylation as well as IH-induced changes in endothelial barrier function. These results demonstrate that IH via ROS-dependent activation of MAP kinases leads to reorganization of cytoskeleton and junction proteins resulting in endothelial barrier dysfunction.