Department of Respiratory Medicine, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, People's Republic of China.
Inflamm Res. 2010 Nov;59(11):949-58. doi: 10.1007/s00011-010-0207-3. Epub 2010 May 8.
The aim of the study was to investigate the role of src-suppressed C kinase substrate (SSeCKS) in the modulation of rat pulmonary microvascular endothelial cells (RPMVEC) permeability elicited by interleukin (IL)-1β and tumor necrosis factor (TNF)-α.
The gene expression of SSeCKS was analyzed by reverse transcription-polymerase chain reaction. Immunoblotting was used to determine the SSeCKS protein expression and the activation of the protein kinase C (PKC) signaling pathway. A RPMVEC monolayer was constructed to determine changes of transendothelial electrical resistance (TER) and FITC-dextran flux (P (d)) across the monolayer. SSeCKS-specific small interfering RNA was transfected into RPMVEC.
IL-1β and TNF-α activated the PKC signaling pathway in RPMVEC, and up-regulated the gene and protein expression of SSeCKS. Depletion of endogenous SSeCKS in RPMVEC significantly attenuated cytokine-induced decrease in TER and increase in P (d), but not to the basal levels. PKC inhibitors also significantly decreased cytokine-induced hyperpermeability and SSeCKS expression.
SSeCKS is involved in the endothelial hyperpermeability induced by IL-1β and TNF-α in inflammatory process.
本研究旨在探讨 src 抑制的 C 激酶底物（SSeCKS）在白细胞介素（IL）-1β和肿瘤坏死因子（TNF）-α诱导的大鼠肺微血管内皮细胞（RPMVEC）通透性变化中的作用。
采用逆转录-聚合酶链反应分析 SSeCKS 的基因表达。免疫印迹法测定 SSeCKS 蛋白表达和蛋白激酶 C（PKC）信号通路的激活。构建 RPMVEC 单层以测定跨内皮电阻（TER）和 FITC-葡聚糖通量（P（d））的变化。将 SSeCKS 特异性小干扰 RNA 转染到 RPMVEC 中。
IL-1β和 TNF-α激活了 RPMVEC 中的 PKC 信号通路，并上调了 SSeCKS 的基因和蛋白表达。RPMVEC 中内源性 SSeCKS 的耗竭显著减弱了细胞因子诱导的 TER 降低和 P（d）增加，但不能达到基础水平。PKC 抑制剂也显著降低了细胞因子诱导的高通透性和 SSeCKS 表达。
SSeCKS 参与了炎症过程中 IL-1β 和 TNF-α诱导的内皮通透性增加。
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