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Mechanisms of tumor necrosis factor-alpha-induced leaks in intestine epithelial barrier.

Abstract：

PURPOSE:The aim of this study was to investigate the signaling mechanisms surrounding changes in tight junction (TJ) and the permeability of human intestinal epithelial cell induced by tumor necrosis factor-alpha (TNF-α). METHODS:To confirm that TNF-α induces epithelial barrier hyperpermeability by disrupting tight junction, Caco-2 cells were exposed to TNF-α, and changes in epithelial permeability (via TER assay), F-actin dynamics (via Rhodamine-phalloidin staining) and tight junction protein expression (via western blot) were monitored. Moreover, to ensure that NF-κB participated in the regulatory mechanisms, Caco-2 cells were transfected with DNMu-IκBα or control plasmids, the above experiments were repeated and the activation effect of TNF-α on NF-κB was detected by luciferase reporter assays. Lastly, we took dominant negative plasmid and knockdown approaches to investigate the potential importance of the NF-κB/myosin light chain kinase (MLCK)/myosin light chain phosphorylation (pMLC) pathways in TNF-a-mediated damage. RESULT:TNF-α could cause NF-κB activation, F-actin rearrangement, tight junction disruption and barrier dysfunction. These effects were alleviated by inhibiting NF-κB. TNF-α induced increase of MLCK transcription and MLC phosphorylation act later than NF-κB activation, which could be suppressed both by inactivating and deleting NF-κB. CONCLUSIONS:TNF-α induces intestinal epithelial cell hyperpermeability by disrupting TJs, in part through MLCK upregulation, in which NF-κB is the positive upstream regulator for MLCK.

journal_name

Cytokine

journal_title

Cytokine

authors

He F,Peng J,Deng XL,Yang LF,Camara AD,Omran A,Wang GL,Wu LW,Zhang CL,Yin F

doi

10.1016/j.cyto.2012.04.008

subject

Has Abstract

pub_date

2012-08-01 00:00:00

pages

264-72

issue

2

eissn

1043-4666

issn

1096-0023

pii

S1043-4666(12)00138-X

journal_volume

59

pub_type

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