Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture

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 h...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Puqiao Lian, Saskia Braber, Soheil Varasteh, Harry J. Wichers, Gert Folkerts
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/853c63bfd71747ac82bfa2e7ed20cb69
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:853c63bfd71747ac82bfa2e7ed20cb69
record_format	dspace
spelling	oai:doaj.org-article:853c63bfd71747ac82bfa2e7ed20cb692021-12-02T17:12:18ZHypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture10.1038/s41598-021-92574-52045-2322https://doaj.org/article/853c63bfd71747ac82bfa2e7ed20cb692021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-92574-5https://doaj.org/toc/2045-2322Abstract 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.Puqiao LianSaskia BraberSoheil VarastehHarry J. WichersGert FolkertsNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Puqiao Lian Saskia Braber Soheil Varasteh Harry J. Wichers Gert Folkerts Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture
description	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.
format	article
author	Puqiao Lian Saskia Braber Soheil Varasteh Harry J. Wichers Gert Folkerts
author_facet	Puqiao Lian Saskia Braber Soheil Varasteh Harry J. Wichers Gert Folkerts
author_sort	Puqiao Lian
title	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_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_sort	hypoxia and heat stress affect epithelial integrity in a caco-2/ht-29 co-culture
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/853c63bfd71747ac82bfa2e7ed20cb69
work_keys_str_mv	AT puqiaolian hypoxiaandheatstressaffectepithelialintegrityinacaco2ht29coculture AT saskiabraber hypoxiaandheatstressaffectepithelialintegrityinacaco2ht29coculture AT soheilvarasteh hypoxiaandheatstressaffectepithelialintegrityinacaco2ht29coculture AT harryjwichers hypoxiaandheatstressaffectepithelialintegrityinacaco2ht29coculture AT gertfolkerts hypoxiaandheatstressaffectepithelialintegrityinacaco2ht29coculture
_version_	1718381443750559744

Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture

Ejemplares similares