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Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture
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 colo...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8222227/ https://www.ncbi.nlm.nih.gov/pubmed/34162953 http://dx.doi.org/10.1038/s41598-021-92574-5 |
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author | Lian, Puqiao Braber, Saskia Varasteh, Soheil Wichers, Harry J. Folkerts, Gert |
author_facet | Lian, Puqiao Braber, Saskia Varasteh, Soheil Wichers, Harry J. Folkerts, Gert |
author_sort | Lian, Puqiao |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8222227 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-82222272021-06-24 Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture Lian, Puqiao Braber, Saskia Varasteh, Soheil Wichers, Harry J. Folkerts, Gert Sci Rep Article 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. Nature Publishing Group UK 2021-06-23 /pmc/articles/PMC8222227/ /pubmed/34162953 http://dx.doi.org/10.1038/s41598-021-92574-5 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Lian, Puqiao Braber, Saskia Varasteh, Soheil Wichers, Harry J. Folkerts, Gert Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture |
title | Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture |
title_full | Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture |
title_fullStr | Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture |
title_full_unstemmed | Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture |
title_short | Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture |
title_sort | hypoxia and heat stress affect epithelial integrity in a caco-2/ht-29 co-culture |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8222227/ https://www.ncbi.nlm.nih.gov/pubmed/34162953 http://dx.doi.org/10.1038/s41598-021-92574-5 |
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