Interleukin-17F-induced pulmonary microvascular endothelial monolayer hyperpermeability via the protein kinase C pathway.

Abstract

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 transendothelial 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.

• Authors: Qing-hai You, Geng-yun Sun, Nan Wang, Ji-long Shen, Yuan Wang
• Journal: The Journal of surgical research (J Surg Res) Vol. 162 Issue 1 Pg. 110-21 (Jul 2010) ISSN: 1095-8673 [Electronic] United States
• PMID: 19577259 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: (c) 2010. Published by Elsevier Inc. All rights reserved.

Chemical References

• A Kinase Anchor Proteins
• Actins
• Akap12 protein, rat
• Cell Cycle Proteins
• Interleukin-17
• Protein Kinase C

Topics

• A Kinase Anchor Proteins (genetics, metabolism)
• Actins (metabolism)
• Animals
• Capillary Permeability
• Cell Cycle Proteins (genetics, metabolism)
• Cells, Cultured
• Cytoskeleton (metabolism)
• Endothelial Cells (metabolism)
• Endothelium, Vascular (metabolism)
• Gene Expression
• Interleukin-17 (metabolism)
• Lung (blood supply)
• Phosphorylation
• Protein Kinase C (metabolism)
• Rats
• Signal Transduction