The role of p38 MAP kinase in hydrogen peroxide mediated endothelial solute permeability
The role of p38 MAP kinase in hydrogen peroxide mediated endothelial solute permeability
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DOI：
10.3109/10623320109165320
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发表时间：
2001-01-01
影响因子：
--
通讯作者：
Alexander, JS
中科院分区：
文献类型：
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作者：
Kevil, CG;Oshima, T;Alexander, JS
目的探讨过氧化氢对内皮细胞溶质通透性增加过程中p38MAPK活性的影响。我们还试图确定p38 MAP激酶介导的内皮细胞结构变化在过氧化氢挑战中的作用。方法在SB202190抑制和不抑制P38 MAP激酶的情况下，测定过氧化氢介导的HUVEC通透性。免疫荧光显微镜观察过氧化氢介导的内皮细胞肌动蛋白、细胞骨架和连接蛋白occludin和ZO-1的重排。结果过氧化氢处理内皮细胞单层90min后，溶质通透性明显增加。SB 202190可显著降低氧化剂介导的p38蛋白激酶的通透性和磷酸化水平。紧密连接蛋白occludin和ZO-1的免疫荧光染色显示，氧化剂攻击导致内皮紧密连接组织的丧失。肌动蛋白细胞骨架罗丹明菲洛定染色显示，过氧化氢刺激应力纤维形成增加，并伴随相邻内皮细胞间隙的形成。在氧化剂攻击过程中抑制p38MAPK可显著减少肌动蛋白应激纤维的形成并阻止缝隙的形成。结论p38MAPK活性参与了过氧化氢介导的通透性、应激纤维的形成和细胞内缝隙的形成。
Objective The purpose of this study was to determine the contribution of p38 MAP kinase activity during hydrogen peroxide mediated increased endothelial solute permeability. We also sought to identify the role of p38 MAP kinase-mediated changes in endothelial cell architecture due to hydrogen peroxide challenge.Methods Hydrogen peroxide mediated permeability of HUVEC was determined with and without inhibition of p38 MAP kinase by SB202190. Hydrogen peroxide mediated rearrangement of the endothelial actin cytoskeleton and junctional proteins occludin and ZO-1 were observed by immunofluorescence microscopy.Results Hydrogen peroxide treatment of endothelial monolayers caused a significant increase in solute permeability over a ninety-minute time period. Oxidant-mediated permeability and phosphorylation of p38 MAP kinase was significantly attenuated by SB 202190. Immunofluorescent staining for the tight junctional proteins occludin and ZO-1 demonstrated that oxidant challenge caused a loss of endothelial tight junction organization. Rhodamine phalloidin staining of the actin cytoskeleton showed that hydrogen peroxide stimulated increased stress fiber formation with concomitant gap formation between adjacent endothelial cells. Inhibition of p38 MAP kinase during oxidant challenge significantly attenuated actin stress fiber formation and prevented gap formation.Conclusions These data demonstrate that p38 MAP kinase activity is involved in hydrogen peroxide mediated permeability, stress fiber formation, and intracellular gap formation.