[A study on increased permeability and morphological changes in actin cytoskeleton and tight junction of vascular endothelial cells induced by tumor necrosis factor-alpha].
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.
Zheng
Yun-jiang
YJ
Surgical Intensive Care Unit, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
Tang
Yao-qing
YQ
Liu
Wei
W
Mao
En-qiang
EQ
Li
Lei
L
Wu
Jun
J
Zhang
Ru-yuan
RY
Zhang
Sheng-dao
SD
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*
D000199:Actins; D014409:Tumor Necrosis Factor-alpha