Interleukin-17F-Induced Pulmonary Microvascular Endothelial Monolayer Hyperpermeability Via the Protein Kinase C Pathway

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.