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Cited by 452Inflammatory Bowel DiseaseBarrier Structure and Function StudiesGastrointestinal motility and disorders

Mechanism of TNF-α modulation of Caco-2 intestinal epithelial tight junction barrier: role of myosin light-chain kinase protein expression

Ma, Thomas Y., Boivin, Michel A., Ye, Dongmei, Pedram, Ali, Said, Hamid M.·2005·American Journal of Physiology-Gastrointestinal and Liver Physiology
DOI 10.1152/ajpgi.00412.2004
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Abstract

TNF-α plays a central role in the intestinal inflammation of various inflammatory disorders including Crohn's disease (CD). TNF-α-induced increase in intestinal epithelial tight junction (TJ) permeability has been proposed as one of the proinflammatory mechanisms contributing to the intestinal inflammation. The intracellular mechanisms involved in the TNF-α-induced increase in intestinal TJ permeability remain unclear. The purpose of this study was to investigate the possibility that the TNF-α-induced increase in intestinal epithelial TJ permeability was regulated by myosin light-chain kinase (MLCK) protein expression, using an in vitro intestinal epithelial model system consisting of the filter-grown Caco-2 intestinal epithelial monolayers. TNF-α (10 ng/ml) produced a time-dependent increase in Caco-2 MLCK expression. The TNF-α increase in MLCK protein expression paralleled the increase in Caco-2 TJ permeability, and the inhibition of the TNF-α-induced MLCK expression (by cycloheximide) prevented the increase in Caco-2 TJ permeability, suggesting that MLCK expression may be required for the increase in Caco-2 TJ permeability. The TNF-α increase in MLCK protein expression was preceded by an increase in MLCK mRNA expression but not an alteration in MLCK protein degradation. Actinomycin-D prevented the TNF-α increase in MLCK mRNA expression and the subsequent increase in MLCK protein expression and Caco-2 TJ permeability, suggesting that the increase in MLCK mRNA transcription led to the increase in MLCK expression. The TNF-α increase in MLCK protein expression was also associated with an increase in Caco-2 MLCK activity. The cycloheximide inhibition of MLCK protein expression prevented the TNF-α increase in MLCK activity and Caco-2 TJ permeability. Moreover, inhibitors of MLCK, Mg 2+-myosin ATPase, and metabolic energy prevented the TNF-α increase in Caco-2 TJ permeability, suggesting that the increase in MLCK activity was required for the TNF-α-induced opening of the Caco-2 TJ barrier. In conclusion, our results indicate for the first time that 1) the TNF-α increase in Caco-2 TJ permeability was mediated by an increase in MLCK protein expression, 2) the increase in MLCK protein expression was regulated by an increase in MLCK mRNA transcription, and 3) the increase in Caco-2 TJ permeability required MLCK protein expression-dependent increase in MLCK activity.

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Crossref · 10.1152/ajpgi.00412.2004

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  • Increased serum concentrations of TNF, IL-1β, and IL-6 are potent inducers of intestinal permeability.

    “Increased serum concentrations of TNF, IL-1β, and IL-6 have also been shown to be potent inducers of intestinal permeability50–53.”

    From Dietary metabolism, the gut microbiome, and heart failure (2019) · cites this paper as [52]

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Authors
Ma, T. Y., Boivin, M. A., Ye, D., Pedram, A., & Said, H. M.
Affiliations
University of New Mexico (US)
ORCID
Venue
American Journal of Physiology-Gastrointestinal and Liver Physiology
Publisher
American Physiological Society
Published
2005-03
Volume
288
Issue
3
Pages
G422-G430
ISSN
  • 0193-1857 (ISSN-L)
  • 1522-1547
DOI
10.1152/ajpgi.00412.2004
OpenAlex
W2136845685
Cited by
452
References
51
Citations by year
2026: 10 · 2025: 12 · 2024: 20 · 2023: 25 · 2022: 21 · 2021: 32 · 2020: 20 · 2019: 22 · 2018: 24 · 2017: 26 · 2016: 32 · 2015: 23 · 2014: 17 · 2013: 23 · 2012: 26
FWCI
9.14
Type
journal article
Language
en
Open access
closed
Locations
  • American Journal of Physiology-Gastrointestinal and Liver Physiology (publishedVersion)
  • PubMed (publishedVersion)
Funders
National Institute of Diabetes and Digestive and Kidney Diseases (R01DK064165, R0-1-DK-64165–01)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Inflammatory Bowel Disease
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
  • Health Sciences › Medicine › Gastroenterology › Gastrointestinal motility and disorders
Keywords
TNF-α, myosin light chain kinase, tight junction permeability, intestinal epithelial barrier, Caco-2 cells, Crohn's disease, intestinal inflammation, cycloheximide, actinomycin D, gene expression regulation
MeSH terms
Antimetabolites, Cell Membrane Permeability, Dactinomycin, Humans, Methionine, Myosin-Light-Chain Kinase, Protein Synthesis Inhibitors, RNA, Messenger, Tumor Necrosis Factor-alpha, Blotting, Western, DNA Primers, Caco-2 Cells, Tight Junctions, Sodium Azide, Reverse Transcriptase Polymerase Chain Reaction, Immunoprecipitation

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