title: Preconditioning of primary human endothelial cells with inflammatory mediators alters the "set point" of the cell.
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)-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.
authors.affiliation: The Center for Vascular Biology Research and Division of Molecular and Vascular Medicine, Beth Israel Deaconess Medical Center, Boston MA 02215, USA.
authors.forename: Youichiro
authors.initials: Y
authors.lastname: Wada
authors.forename: Hasan
authors.initials: H
authors.lastname: Otu
authors.forename: Shengqian
authors.initials: S
authors.lastname: Wu
authors.forename: Md Ruhul
authors.initials: MR
authors.lastname: Abid
authors.forename: Hitomi
authors.initials: H
authors.lastname: Okada
authors.forename: Towia
authors.initials: T
authors.lastname: Libermann
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Kodama
authors.forename: Shu-Ching
authors.initials: SC
authors.lastname: Shih
authors.forename: Takashi
authors.initials: T
authors.lastname: Minami
authors.forename: William C
authors.initials: WC
authors.lastname: Aird
mesh_terms: D021581:Active Transport, Cell Nucleus; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D017931:DNA Primers; D004305:Dose-Response Relationship, Drug; D019040:E-Selectin / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000473:pathology; D005334:Fever / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors / Q000096:biosynthesis; D000860:Hypoxia / Q000378:metabolism; D020672:I-kappa B Proteins / Q000378:metabolism; D007249:Inflammation; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D008954:Models, Biological; D019001:Nitric Oxide Synthase / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation; D010982:Platelet-Derived Growth Factor / Q000378:metabolism; D021381:Protein Transport; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012260:Ribonucleases / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D013917:Thrombin / Q000737:chemistry / Q000378:metabolism; D013997:Time Factors; D051996:Transcription Factor RelA / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: D003470:Culture Media; D017931:DNA Primers; D019040:E-Selectin; D018960:Hyaluronan Receptors; D020672:I-kappa B Proteins; D008070:Lipopolysaccharides; D010982:Platelet-Derived Growth Factor; C498143:RELA protein, human; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; C075278:platelet-derived growth factor A; D018799:Intercellular Adhesion Molecule-1; D019001:Nitric Oxide Synthase; D012260:Ribonucleases; D013917:Thrombin