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

FASEB J. 2005 Nov;19(13):1914-6. doi: 10.1096/fj.05-4037fje. Epub 2005 Sep 19.


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)-alpha, 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-alpha 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-kappaB, an effect that was significantly blunted by pretreatment of cells with TNF-alpha and LPS. Consistent with these results, TNF-alpha and LPS attenuated the effect of thrombin on IkappaB phosphorylation, total cytoplasmic IkappaB, and nuclear translocation of p65 NF-kappaB. The inhibitory effect of TNF-alpha 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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Active Transport, Cell Nucleus
  • Cells, Cultured
  • Culture Media / metabolism
  • Cytoplasm / metabolism
  • DNA Primers
  • Dose-Response Relationship, Drug
  • E-Selectin / metabolism
  • Endothelial Cells / cytology*
  • Endothelium, Vascular / cytology*
  • Endothelium, Vascular / pathology
  • Fever / metabolism
  • Humans
  • Hyaluronan Receptors / biosynthesis
  • Hypoxia / metabolism
  • I-kappa B Proteins / metabolism
  • Inflammation*
  • Intercellular Adhesion Molecule-1 / metabolism
  • Lipopolysaccharides / metabolism
  • Models, Biological
  • Nitric Oxide Synthase / metabolism
  • Oligonucleotide Array Sequence Analysis
  • Phosphorylation
  • Platelet-Derived Growth Factor / metabolism
  • Protein Transport
  • Reverse Transcriptase Polymerase Chain Reaction
  • Ribonucleases / chemistry
  • Ribonucleases / metabolism
  • Signal Transduction
  • Thrombin / chemistry
  • Thrombin / metabolism
  • Time Factors
  • Transcription Factor RelA / metabolism
  • Tumor Necrosis Factor-alpha / metabolism
  • Umbilical Veins / cytology*
  • Vascular Cell Adhesion Molecule-1 / metabolism


  • Culture Media
  • DNA Primers
  • E-Selectin
  • Hyaluronan Receptors
  • I-kappa B Proteins
  • Lipopolysaccharides
  • Platelet-Derived Growth Factor
  • RELA protein, human
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha
  • Vascular Cell Adhesion Molecule-1
  • platelet-derived growth factor A
  • Intercellular Adhesion Molecule-1
  • Nitric Oxide Synthase
  • Ribonucleases
  • Thrombin