Preconditioning of primary human endothelial cells with inflammatory mediators alters the “set point” of the cell

Authors

Youichiro Wada, Beth Israel Deaconess Medical Center
Hasan H. Otu, Beth Israel Deaconess Medical CenterFollow
Shengqian Wu, Beth Israel Deaconess Medical Center
Md. Ruhul Abid, Beth Israel Deaconess Medical Center
Hitomi Okada, Beth Israel Deaconess Medical Center
Towia Libermann, Beth Israel Deaconess Medical Center
Tatsuhiko Kodama, University of Tokyo
Shu-Ching Shih, Beth Israel Deaconess Medical Center
Takashi Minami, University of Tokyo
William C. Aird, Beth Israel Deaconess Medical CenterFollow

Document Type

Article

Date of this Version

2005

Citation

FASEB J. 2005 November ; 19(13): 1914–1916

Comments

Copyright 2005 FASEB

doi:10.1096/fj.05-4037fje

Abstract

Endothelial cells are highly sensitive to changes in the extracellular milieu. Sepsis results in activation of inflammatory and coagulation pathways. We hypothesized that sepsis-associated mediators may alter the response capacity (so-called “set point”) of endothelial cells. Human umbilical vein endothelial cells (HUVEC) were preincubated in the presence or absence of tumor necrosis factor (TNF)-α, lipopolysaccharide (LPS), hypoxia, hyperthermia, and/or high glucose; treated with or without thrombin for 4 h; and then processed for RNase protection assays of selected activation markers. Priming with TNF-α and LPS significantly inhibited thrombin-mediated induction of vascular cell adhesion molecule-1, intercellular adhesion molecule-1, tissue factor, and E-selectin, but not platelet-derived growth factor-A or CD44. In electrophoretic mobility shift assays, thrombin-treated HUVEC demonstrated inducible binding of p65 NF-κB, an effect that was significantly blunted by pretreatment of cells with TNF-α and LPS. Consistent with these results, TNF-α and LPS attenuated the effect of thrombin on IκB phosphorylation, total cytoplasmic IκB, and nuclear translocation of p65 NF-κB. The inhibitory effect of TNF-α on thrombin signaling persisted for up to 24 h following removal of the cytokine. Taken together, these data suggest that inflammatory mediators prime endothelial cells to modulate subsequent thrombin response.