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

Cytokine
Fang HeFei Yin

Abstract

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). 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. 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 phosphoryl...Continue Reading

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Citations

Nov 19, 2013·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Songwei YangHua Yang
Sep 10, 2015·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Qianhuining KuangYuhui Ni
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Aug 6, 2013·Cytokine·Jaione BarrenetxeMaria Pilar Lostao
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Apr 6, 2019·Digestive Diseases and Sciences·Mingyue YangXiaolan Zhang
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