Tumor Necrosis Factor Alpha Effects on the Porcine Intestinal Epithelial Barrier Include Enhanced Expression of TNF Receptor 1

Tumor necrosis factor alpha (TNFα) has been shown to impair the intestinal barrier, inducing and maintaining inflammatory states of the intestine. The aim of the current study was to analyze functional, molecular and regulatory effects of TNFα in a newly established non-transformed jejunal enterocyt...

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Bibliographic Details
Published in:International journal of molecular sciences 2021-08, Vol.22 (16), p.8746
Main Authors: Droessler, Linda, Cornelius, Valeria, Markov, Alexander G., Amasheh, Salah
Format: Article
Language:English
Subjects:
Apoptosis
Cell culture
claudins
Cytokines
Electrical resistivity
epithelial barrier
Epithelium
Experiments
Immunofluorescence
Inflammation
Inflammatory bowel disease
Intestine
IPEC-J2
Kinases
Localization
Mannitol
ML-7
Myosin
Myosin-light-chain kinase
Necrosis
Permeability
Protein expression
Proteins
tight junction
TNFα
Tumor necrosis factor receptors
Tumor necrosis factor-TNF
Tumor necrosis factor-α
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Summary:Tumor necrosis factor alpha (TNFα) has been shown to impair the intestinal barrier, inducing and maintaining inflammatory states of the intestine. The aim of the current study was to analyze functional, molecular and regulatory effects of TNFα in a newly established non-transformed jejunal enterocyte model, namely IPEC-J2 monolayers. Incubation with 1000 U/mL TNFα induced a marked decrease in transepithelial electrical resistance (TEER), and an increase in permeability for the paracellular flux marker [3H]-D-mannitol compared to controls. Immunoblots revealed a significant decrease in tight junction (TJ) proteins occludin, claudin-1 and claudin-3. Moreover, a dose-dependent increase in the TNF receptor (TNFR)-1 was detected, explaining the exponential nature of pro-inflammatory effects, while TNFR-2 remained unchanged. Recovery experiments revealed reversible effects after the removal of the cytokine, excluding apoptosis as a reason for the observed changes. Furthermore, TNFα signaling could be inhibited by the specific myosin light chain kinase (MLCK) blocker ML-7. Results of confocal laser scanning immunofluorescence microscopy were in accordance with all quantitative changes. This study explains the self-enhancing effects of TNFα mediated by MLCK, leading to a differential regulation of TJ proteins resulting in barrier impairment in the intestinal epithelium.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms22168746