In Vitro Evaluation of the Effects of Commercial Prebiotic GOS and FOS Products on Human Colonic Caco–2 Cells

Abstract

Prebiotic oligosaccharides are widely used as human and animal feed additives for their beneficial effects on the gut microbiota. However, there are limited data to assess the direct effect of such functional foods on the transcriptome of intestinal epithelial cells. The purpose of this study is to describe the differential transcriptomes and cellular pathways of colonic cells directly exposed to galacto-oligosaccharides (GOS) and fructo-oligosaccharides (FOS). We have examined the differential gene expression of polarized Caco–2 cells treated with GOS or FOS products and their respective mock-treated cells using mRNA sequencing (RNA-seq). A total of 89 significant differentially expressed genes were identified between GOS and mock-treated groups. For FOS treatment, a reduced number of 12 significant genes were observed to be differentially expressed relative to the control group. KEGG and gene ontology functional analysis revealed that genes up-regulated in the presence of GOS were involved in digestion and absorption processes, fatty acids and steroids metabolism, potential antimicrobial proteins, energy-dependent and -independent transmembrane trafficking of solutes and amino acids. Using our data, we have established complementary non-prebiotic modes of action for these frequently used dietary fibers.

Galactooligosaccharide Treatment Alleviates DSS-Induced Colonic Inflammation in Caco-2 Cell Model

“…From all our data we can speculate that in DSS treated cells, GOS was able to counteract the inflammatory status and the membrane barrier damage induced by inflammatory cytokines that reverted to control level. The ability of GOS to regulate immune response was previously suggested, as it has been shown to be effective in modulating cytokine secretion in intestinal cells through TLR4 binding ( 56 ), as well as through a direct effect on intestinal cell transcriptome by modulating the expression of several genes, including some involved in antimicrobial activity and inflammatory response ( 36 ). According to previous studies, our results strongly suggest that a specific prebiotic can have a direct effect on the regulation of inflammation.…”

“…In in vitro models, GOS has also been shown to modulate the epithelial barrier function by inducing differentiation and epithelial wound repair, and by promoting the growth of specific gut bacteria, associated with changes in SCFA profiles ( 35 ). The effect of GOS on epithelial cells has been further confirmed by a transcriptomic analysis performed on Caco-2 cells, showing that GOS was able to modulate the expression of several genes implicated in digestion and transepithelial transport, which contribute to intestinal cell integrity and function ( 36 ).…”

Microbiota independent effects of oligosaccharides on Caco-2 cells -A semi-targeted metabolomics approach using DI-FT-ICR-MS coupled with pathway enrichment analysis

“…Medium was changed every second day. After 20 days, cells were treated in accordance with the experiments of Lafontaine et al (2020) for 24 h with HG, GM, LGM, or (as a control) with a sugar mixture (galactose, glucose and lactose [1/1/1, w/w/w]) and at a concentration of 25 mg ml −1 ( Zenhom et al, 2011 ). After this incubation, cells were visually inspected, medium was removed, cells were washed twice with ice-cold deionized H 2 O, snap frozen in liquid nitrogen to quench cell metabolism, and stored at −80°C.…”

Impact of dietary fibers in infant formulas on gut microbiota and the intestinal immune barrier

“…70,71 Many studies have demonstrated that microorganisms and dietary components influence the regulation of the TJ protein expression. 72,73 Pattern recognition receptors (PRRs), which are widely expressed by the intestinal epithelial cells and intestinal immune cells, play an important role in the regulation of barrier integrity and TJ expression. 74,75 Toll-like receptors (TLRs) are the most well-studied PRRs, among which TLR2 and TLR4 are the best studied due to their interaction with gut bacteria and dietary components and their ability to mediate epithelial barrier integrity.…”

“…129 In another study, exposure of Caco-2 cells to GOS of DP2-7 (1.4% w/v) also showed the ability to improve the integrity of the tight junction via increasing the trans-epithelial electrical resistance (TER). 73 More recently, the transcriptome of epithelial Caco-2 cells was fully characterized after stimulation with GOS and the hMO sialyllactose (SL) and the effects on the epithelial barrier function were studied by investigating cellular pathways involved in cell proliferation, differentiation, and re-epithelialization. GOS modulated 63 pathways of the epithelial barrier transcriptomes, among which the top 4 most significantly modulated pathways were influenced in the same way by the hMO SL, indicating a similarity in function between GOS and SL.…”