Role of src-suppressed C kinase substrate in rat pulmonary microvascular endothelial hyperpermeability stimulated by inflammatory cytokines

• Published in: Inflammation research 2010-11, Vol.59 (11), p.949-958
• Main Authors: You, Qing-Hai, Sun, Geng-Yun, Wang, Nan, Chen, Shan, Luo, Qing-Li
• Format: Article
• Language: English
• Subjects: A Kinase Anchor Proteins - genetics; A Kinase Anchor Proteins - metabolism; Allergology; Animals; Biomedical and Life Sciences; Biomedicine; Capillary Permeability; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cytokine; Cytokines - metabolism; Dermatology; Endothelial Cells - cytology; Endothelial Cells - metabolism; Immunology; Interleukin-1beta - immunology; Lung - blood supply; Microcirculation; Neurology; Original Research Paper; permeability; Pharmacology/Toxicology; protein kinase C; Protein Kinase C - metabolism; Pulmonary microvascular endothelial cells; Rats; Rats, Wistar; Rheumatology; Signal Transduction - immunology; Src-suppressed C kinase substrate; Tumor Necrosis Factor-alpha - immunology
• ISSN: 1023-3830; 1420-908X
• DOI: 10.1007/s00011-010-0207-3

Objective 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. Methods 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. Results 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. Conclusions SSeCKS is involved in the endothelial hyperpermeability induced by IL-1β and TNF-α in inflammatory process.