title: [A study on increased permeability and morphological changes in actin cytoskeleton and tight junction of vascular endothelial cells induced by tumor necrosis factor-alpha].
abstract: OBJECTIVE
  To study the effect of human tumor necrosis factor-alpha (TNF-alpha) on permeability of human vascular endothelial cell (EA.hy926) monolayer and its mechanism.
  
  
  METHODS
  5, 10, 20 microg/L TNF-alpha was respectively added to the cultured endothelial cell monolayer for 24 hours, or 10 microg/L TNF-alpha for 6, 12, 24 hours. Human vascular endothelial cell (EA.hy926) monolayer permeability was measured by detecting fluorescence intensity of fluorescein isothiocyanate (FITC) labeled dextran. Immunofluorescence and laser confocal microscopy were used to assess vascular endothelial actin cytoskeleton (F-actin) and tight junction protein (ZO-1) distribution. Western blotting was used to assess ZO-1 expression.
  
  
  RESULTS
  Compared with control group, TNF-alpha significantly increased endothelial permeability and induced F-actin redistribution and stress fiber formation with ZO-1 derangement. Gaps increased obviously between endothelial cells. Furthermore, Western blotting showed that TNF-alpha reduced ZO-1 expression in a dose- and time-dependent manner.
  
  
  CONCLUSIONS
  TNF-alpha increased endothelial cell permeability by damaging integrity of endothelial barrier function.
authors.affiliation: Surgical Intensive Care Unit, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
authors.forename: Yun-jiang
authors.initials: YJ
authors.lastname: Zheng
authors.forename: Yao-qing
authors.initials: YQ
authors.lastname: Tang
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: En-qiang
authors.initials: EQ
authors.lastname: Mao
authors.forename: Lei
authors.initials: L
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Wu
authors.forename: Ru-yuan
authors.initials: RY
authors.lastname: Zhang
authors.forename: Sheng-dao
authors.initials: SD
authors.lastname: Zhang
mesh_terms: D000199:Actins / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D006801:Humans; D019108:Tight Junctions / Q000187:drug effects / Q000648:ultrastructure*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D000199:Actins; D014409:Tumor Necrosis Factor-alpha