Interleukin-17F-Induced Pulmonary Microvascular Endothelial Monolayer Hyperpermeability Via the Protein Kinase C Pathway
Interleukin-17F-Induced Pulmonary Microvascular Endothelial Monolayer Hyperpermeability Via the Protein Kinase C Pathway
复制标题
Interleukin-17F 通过蛋白激酶 C 途径诱导肺微血管内皮单层通透性过高
DOI：
10.1016/j.jss.2009.01.019
复制
发表时间：
2010-07-01
影响因子：
2.2
通讯作者：
Wang, Yuan
中科院分区：
文献类型：
--
作者：
You, Qing-hai;Sun, Geng-yun;Wang, Yuan
背景白细胞介素（IL）-17 F参与肺部炎症，但IL-17 F对内皮通透性及其信号通路的影响仍然不清楚。本研究旨在探讨IL-17 F对内皮细胞的影响，并评估蛋白激酶C（PKC）和src抑制的C激酶底物（SSeCKS）在这一过程中的作用。构建大鼠肺微血管内皮单层以确定渗透性的变化，所述渗透性的变化通过FITC-葡聚糖和Hank溶液穿过单层的通量以及有或没有IL-17 F和PKC抑制剂的跨内皮电阻来测量。使用FITC-鬼笔环肽对丝状肌动蛋白（F-肌动蛋白）的其他单层进行染色。用逆转录-聚合酶链反应分析SSeCKS基因的表达。免疫印迹和免疫沉淀法检测SSeCKS蛋白的变化。IL-17 F以剂量和时间依赖性方式增加内皮单层通透性。F-肌动蛋白染色显示，通透性的变化伴随着细胞骨架的重组。在PKC抑制剂的存在下，IL-17 F诱导的通透性增高和F-肌动蛋白的重组被减弱。IL-17 F刺激后，SSeCKS基因和蛋白表达均显著升高。SSeCKS磷酸化的过程遵循反映由IL-17 F诱导的高通透性的时间过程的时间过程。IL-17 F诱导的SSeCKS磷酸化在PKC抑制剂预处理后被废除。IL-17 F处理后，SSeCKS从胞浆移位到胞膜，与细胞骨架的结合显著增加。IL-17 F是内皮通透性增加的重要介质。PKC和SSeCKS是IL-17 F诱导的高通透性所必需的整合信号传导组分。皇冠版权所有（C）2010由爱思唯尔公司出版。All rights reserved.
Background. Interleukin (IL)-17F is involved in lung inflammation, but the effect of IL-17F on endothelial permeability and its signaling pathway remain ill-defined. The current study sought to investigate the effect of IL-17F on endothelium and assess the role of protein kinase C (PKC) and src-suppressed C kinase substrate (SSeCKS) in this process.Methods. Rat pulmonary microvascular endothelial monolayers were constructed to determine changes of permeability as measured by means of FITC-dextran and Hank's solution flux across monolayers and trans-endothelial electrical resistance with or without IL-17F and PKC inhibitors. Additional monolayers were stained using FITC-phalloidin for filamentous actin (F-actin). The gene expression of SSeCKS was analyzed by the reverse transcription-polymerase chains. Alterations of SSeCKS protein were investigated by immunoblotting and immunoprecipitation.Results. IL-17F increased endothelial monolayer permeability in a dose- and time-dependent manner. F-actin staining revealed that permeability changes were accompanied by reorganization of cytoskeleton. In the presence of PKC inhibitors, the IL-17F-induced hyperpermeability and reorganization of F-actin were attenuated. The gene and protein expression of SSeCKS were conspicuously elevated after IL-17F challenge. The process of SSeCKS phosphorylation followed a time course that mirrored the time course of hyperpermeability induced by IL-17F. IL-17F-induced SSeCKS phosphorylation was abrogated after PKC inhibitors pretreatment. The translocation of SSeCKS from the cytosol to the membrane and a significant increase in the SSeCKS association with the cytoskeleton were found after IL-17F treatment.Conclusions. IL-17F is an important mediator of increased endothelial permeability. PKC and SSeCKS are integral signaling components essential for IL-17F-induced hyperpermeability. Crown Copyright (C) 2010 Published by Elsevier Inc. All rights reserved.