Abstract
The role of reactive oxygen species (ROS) in osmotic stress, dextran sulfate sodium (DSS) and cyclic stretch-induced tight junction (TJ) disruption was investigated in Caco-2 cell monolayers in vitro and restraint stress-induced barrier dysfunction in mouse colon in vivo. Live cell imaging showed that osmotic stress, cyclic stretch and DSS triggered rapid production of ROS in Caco-2 cell monolayers, which was blocked by depletion of intracellular Ca2+ by 1,2-bis-(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid. Knockdown of CaV1.3 or TRPV6 channels blocked osmotic stress and DSS-induced ROS production and attenuated TJ disruption and barrier dysfunction. N-Acetyl L-cysteine (NAC) and L-NG-Nitroarginine methyl ester (L-NAME) blocked stress-induced TJ disruption and barrier dysfunction. NAC and L-NAME also blocked stress-induced activation of c-Jun N-terminal kinase (JNK) and c-Src. ROS was colocalized with the mitochondrial marker in stressed cells. Cyclosporin A blocked osmotic stress and DSS-induced ROS production, barrier dysfunction, TJ disruption and JNK activation. Mitochondria-targeted Mito-TEMPO blocked osmotic stress and DSS-induced barrier dysfunction and TJ disruption. Chronic restraint stress in mice resulted in the elevation of intracellular Ca2+, activation of JNK and c-Src, and disruption of TJ in the colonic epithelium. Furthermore, corticosterone administration induced JNK and c-Src activation, TJ disruption and protein thiol oxidation in colonic mucosa. The present study demonstrates that oxidative stress is a common signal in the mechanism of TJ disruption in the intestinal epithelium by different types of cellular stress in vitro and bio behavioral stress in vivo.
Original language | English |
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Pages (from-to) | 731-749 |
Number of pages | 19 |
Journal | Biochemical Journal |
Volume | 474 |
Issue number | 5 |
DOIs | |
State | Published - Mar 1 2017 |
Bibliographical note
Publisher Copyright:© 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.
Funding
The present study was supported by the National Institute of Health grants [R01-DK55532, R01-AA12307, CA109298 and CA083639].
Funders | Funder number |
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National Institutes of Health (NIH) | R01-AA12307, CA109298, R01-DK55532 |
National Childhood Cancer Registry – National Cancer Institute | P50CA083639 |
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Cell Biology