Butyrate Does Not Protect Against Inflammation-induced Loss of Epithelial Barrier Function and Cytokine Production in Primary Cell Monolayers From Patients With Ulcerative Colitis
Open Access
- 28 March 2019
- journal article
- research article
- Published by Oxford University Press (OUP) in Journal of Crohn's and Colitis
- Vol. 13 (10), 1351-1361
- https://doi.org/10.1093/ecco-jcc/jjz064
Abstract
In vitro studies using immortalised cancer cell lines showed that butyrate has an overall positive effect on epithelial barrier integrity, but the physiological relevance of cancer cell lines is limited. We developed epithelial monolayers from human tissue samples of patients with ulcerative colitis [UC] to assess the effect of butyrate on epithelial barrier function. A protocol to establish monolayers from primary epithelial cells of UC patients [n = 10] and non-UC controls [n = 10] was optimised. The monolayers were treated with 8 mM sodium butyrate ± tumour necrosis factor alpha [TNFα] and type II interferon [IFNγ] for 48 h. Changes in transepithelial electrical resistance were monitored. Barrier gene expression levels were measured. Inflammatory proteins in the supernatant of the cells were quantified with OLINK. We demonstrated that primary monolayer cultures can be grown within 1 week of culture with robust resistance values and polarised tight junction expression. Butyrate treatment of the cultures increased resistance but was detrimental in combination with TNFα and IFNγ. The combined treatment further induced even higher IL8 mRNA and inflammatory protein secretion than for the inflammatory mediators alone. The observed effects were similar in cultures from patients and non-UC controls, suggesting that there were no patient-specific responses responsible for these findings. We found that butyrate does not protect against inflammation-induced barrier dysfunction and even worsens its effects in primary epithelial monolayers of UC patients and controls. The basic mechanisms of butyrate should therefore be reconsidered in future studies, in particular in patients with active inflammation and pre-existing barrier defects as is known for UC.Keywords
Funding Information
- European Research Council [ERC] (ERC-2015-AdG, 694679)
This publication has 63 references indexed in Scilit:
- A decrease of the butyrate-producing speciesRoseburia hominisandFaecalibacterium prausnitziidefines dysbiosis in patients with ulcerative colitisGut, 2013
- Amelioration of IFN-γ and TNF-α-Induced Intestinal Epithelial Barrier Dysfunction by Berberine via Suppression of MLCK-MLC Phosphorylation Signaling PathwayPLOS ONE, 2013
- Alternative functional in vitro models of human intestinal epitheliaFrontiers in Pharmacology, 2013
- Host–microbe interactions have shaped the genetic architecture of inflammatory bowel diseaseNature, 2012
- Butyrate Enhances the Intestinal Barrier by Facilitating Tight Junction Assembly via Activation of AMP-Activated Protein Kinase in Caco-2 Cell MonolayersJournal of Nutrition, 2009
- Faecalibacterium prausnitzii is an anti-inflammatory commensal bacterium identified by gut microbiota analysis of Crohn disease patientsProceedings of the National Academy of Sciences, 2008
- Cytokine regulation of tight junctionsBiochimica et Biophysica Acta (BBA) - Biomembranes, 2008
- A new mathematical model for relative quantification in real-time RT-PCRNucleic Acids Research, 2001
- Interferon γ induces differential upregulation of α and β chemokine secretion in colonic epithelial cell linesGut, 1998
- Rectal irrigation with short-chain fatty acids for distal ulcerative colitisDigestive Diseases and Sciences, 1991