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Preconditioning of primary human endothelial cells with inflammatory mediators alters the “set point” of the cell
Author(s) -
Wada Youichiro,
Otu Hasan,
Wu Shengqian,
Abid Md. Ruhul,
Okada Hitomi,
Libermann Towia,
Kodama Tatsuhiko,
Shih ShuChing,
Minami Takashi,
Aird William C.
Publication year - 2005
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.05-4037fje
Subject(s) - thrombin , tissue factor , tumor necrosis factor alpha , endothelial stem cell , microbiology and biotechnology , umbilical vein , cytokine , lipopolysaccharide , cell adhesion molecule , chemistry , immunology , biology , medicine , coagulation , biochemistry , platelet , in vitro
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.