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Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture.
- Source :
-
Scientific reports [Sci Rep] 2021 Jun 23; Vol. 11 (1), pp. 13186. Date of Electronic Publication: 2021 Jun 23. - Publication Year :
- 2021
-
Abstract
- Hypoxia and hyperthermia, which can be induced by high environmental temperature or strenuous exercise, are two common stressors that affect intestinal epithelial integrity and lead to multiple clinical symptoms. In this study, we developed an in-vitro intestinal monolayer model using two human colonic epithelial cell lines, Caco-2 and HT-29, co-cultured in Transwell inserts, and investigated the effects of heat treatment and/or hypoxia on the epithelial barrier function. The monolayer with a ratio of 9:1 (Caco-2:HT-29) showed high trans-epithelial electrical resistance (TEER), low Lucifer Yellow permeability and high mucin production. Hyperthermia and/or hypoxia exposure (2 h) triggered heat shock and oxidative stress responses. HSP-70 and HSF-1 protein levels were up-regulated by hyperthermia, which were further enhanced when hyperthermia was combined with hypoxia. Increased HIF-1α protein expression and Nrf2 nuclear translocation was only caused by hypoxia. Hyperthermia and/or hypoxia exposure disrupted the established monolayer by increasing paracellular permeability, decreasing ZO-1, claudin-3 and occludin protein/mRNA expression, while enhancing E-cadherin protein expression. Tight junction protein distribution in the monolayer was also modulated by the hyperthermia and/or hypoxia exposure. In addition, transcription levels of mucin genes, MUC-2 and MUC-5AC, were increased after 2 h of hyperthermia and/or hypoxia exposure. In conclusion, this Caco-2/HT-29 cell model is valid and effective for studying detrimental effects of hyperthermia and/or hypoxia on intestinal barrier function and related heat shock and oxidative stress pathways and can be used to investigate possible interventions to reverse hyperthermia and/or hypoxia-induced intestinal epithelial injury.
- Subjects :
- Adenocarcinoma pathology
Adenocarcinoma, Mucinous pathology
Cell Line, Tumor
Coculture Techniques
Colonic Neoplasms pathology
Coloring Agents
Electric Impedance
Gene Expression Regulation, Neoplastic
Humans
Intercellular Junctions
Isoquinolines
Mucins biosynthesis
Neoplasm Proteins biosynthesis
Neoplasm Proteins genetics
Oxidative Stress
RNA, Messenger biosynthesis
RNA, Messenger genetics
RNA, Neoplasm biosynthesis
RNA, Neoplasm genetics
Transcription, Genetic
Cell Hypoxia genetics
Cell Hypoxia physiology
Enterocytes physiology
Goblet Cells physiology
Heat-Shock Response genetics
Heat-Shock Response physiology
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 11
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 34162953
- Full Text :
- https://doi.org/10.1038/s41598-021-92574-5