title: Tumor necrosis factor-related apoptosis-inducing ligand promotes microvascular endothelial cell hyperpermeability through phosphatidylinositol 3-kinase pathway.
abstract: BACKGROUND
  Microvascular hyperpermeability that occurs in hemorrhagic shock and burn trauma is regulated by the apoptotic signaling pathway. We hypothesized that tumor necrosis factor-α (TNF-α)-related apoptosis-inducing ligand (TRAIL) would promote hyperpermeability directly or by interacting with other signaling pathways.
  
  
  METHODS
  Rat lung microvascular endothelial cells (RLMECs) grown on Transwell membranes (Corning Life Sciences, Lowell, MA) were treated with recombinant human TRAIL (10, 50, and 100 ng/mL) for 6 hours or TRAIL (100 ng/mL) + LY294002 (a PI3K inhibitor; 20 μmol/L), Z-DEVD-FMK (a caspase-3 inhibitor; 10 μmol/L), or the inhibitors alone. Fluorescein isothiocyanate (FITC)-albumin flux was an indicator of permeability. Caspase-3 activity was measured fluorometrically. Adherens junction integrity was studied using β-catenin immunofluorescence.
  
  
  RESULTS
  TRAIL + LY294002, but not TRAIL alone, induced monolayer hyperpermeability (P < .05), and caspase-3 activity (P < .05), and disrupted the adherens junctions. Z-DEVD-FMK attenuated hyperpermeability and protected the adherens junctions.
  
  
  CONCLUSIONS
  TRAIL-induced microvascular hyperpermeability is phosphatidylinositol 3-kinase (PI3K)-dependent and may be mediated by caspase-3 cleavage of the endothelial adherens junctional complex.
authors: Department of Surgery, Texas A&M Health Science Center College of Medicine and Scott & White Memorial Hospital, 702 SW H.K. Dodgen Loop, Temple, TX 76504, USA.
authors: Hayden W
authors: HW
authors: Stagg
authors: Kanika A
authors: KA
authors: Bowen
authors: Devendra A
authors: DA
authors: Sawant
authors: Miguel
authors: M
authors: Rodriguez
authors: Binu
authors: B
authors: Tharakan
authors: Ed W
authors: EW
authors: Childs
mesh_terms: D022005:Adherens Junctions / Q000502:physiology*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002199:Capillary Permeability / Q000502:physiology*; D053148:Caspase 3 / Q000378:metabolism*; D002478:Cells, Cultured; D002867:Chromones / Q000378:metabolism; D042783:Endothelial Cells / Q000502:physiology*; D006801:Humans; D009025:Morpholines / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism
chemical_list: D015415:Biomarkers; D002867:Chromones; C495268:Ctnnb1 protein, rat; D009025:Morpholines; D009842:Oligopeptides; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011994:Recombinant Proteins; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; C110772:benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone; D051176:beta Catenin; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D053148:Caspase 3