Hollebeeck S.,Institute Des Science Of La Vie And Uclouvain |
Winand J.,Institute Des Science Of La Vie And Uclouvain |
Herent M.-F.,Leuven Drug Research Institute LDRI and UCLouvain |
During A.,Institute Des Science Of La Vie And Uclouvain |
And 3 more authors.
Food and Function | Year: 2012
This study aimed at evaluating the anti-inflammatory properties of a pomegranate fruit husk (PomH) polyphenolic extract, rich in punicalagin, using Caco-2 cells, an in vitro model of human intestinal epithelium. Differentiated cells in bicameral inserts were pretreated or not with a PomH extract or punicalagin, as reference, at the apical side, representing the intestinal lumen. Inflammation was then induced with a cocktail of cytokines (Il-1β, TNFα and IFNγ) and LPS. After 24 h incubation, 3 pro-inflammatory markers, i.e., interleukin (IL)-6, IL-8 and monocyte chemoattractant protein (MCP)-1, were assayed both at their gene transcription (qRT-PCR) and secretion (ELISA) levels. As previously described, the pro-inflammatory cocktail significantly stimulated these 3 markers, at the gene transcript and secretion levels. In inflamed cells, a significant down-regulation of the transcription of the genes encoding IL-6 and MCP-1 was observed in the presence of the PomH extract or punicalagin, while IL-8 transcription was unaffected. Both treatments also decreased the amounts of the 3 proteins with dose-response effects, but only in the apical compartment. A lowered ELISA response was also observed when either IL-6, IL-8 or MCP-1 were mixed with punicalagin in a cell-free culture medium, indicating a direct molecular interaction. In conclusion, the punicalagin-rich PomH extract tested showed anti-inflammatory properties in the Caco-2 in vitro intestinal model. It acted both on the pro-inflammatory gene transcription and protein levels, the later phenomenon being possibly due to a direct molecular trapping. These data suggest that pomegranate husk could be an interesting natural source contributing to prevent intestinal chronic inflammation. © The Royal Society of Chemistry 2012.
De Walle J.V.,Institute Des Science Of La Vie And Uclouvain |
Sergent T.,Institute Des Science Of La Vie And Uclouvain |
Piront N.,University of Namur |
Toussaint O.,University of Namur |
And 2 more authors.
Toxicology and Applied Pharmacology | Year: 2010
Deoxynivalenol (DON), one of the most common mycotoxin contaminants of raw and processed cereal food, adversely affects the gastrointestinal tract. Since DON acts as a protein synthesis inhibitor, the constantly renewing intestinal epithelium could be particularly sensitive to DON. We analyzed the toxicological effects of DON on intestinal epithelial protein synthesis and barrier integrity. Differentiated Caco-2 cells, as a widely used model of the human intestinal barrier, were exposed to realistic intestinal concentrations of DON (50, 500 and 5000ng/ml) during 24h. DON caused a concentration-dependent decrease in total protein content associated with a reduction in the incorporation of [3H]-leucine, demonstrating its inhibitory effect on protein synthesis. DON simultaneously increased the paracellular permeability of the monolayer as reflected through a decreased transepithelial electrical resistance associated with an increased paracellular flux of the tracer [3H]-mannitol. A concentration-dependent reduction in the expression level of the tight junction constituent claudin-4 was demonstrated by Western blot, which was not due to diminished transcription, increased degradation, or NF-κB, ERK or JNK activation, and was also observed for a tight junction independent protein, i.e. intestinal alkaline phosphatase. These results demonstrate a dual toxicological effect of DON on differentiated Caco-2 cells consisting in an inhibition of protein synthesis as well as an increase in monolayer permeability, and moreover suggest a possible link between them through diminished synthesis of the tight junction constituent claudin-4. © 2010 Elsevier Inc.
Thiry C.,Institute Des Science Of La Vie And Uclouvain |
Ruttens A.,Coda Research |
Pussemier L.,Coda Research |
Schneider Y.-J.,Institute Des Science Of La Vie And Uclouvain
British Journal of Nutrition | Year: 2013
A range of Se species has been shown to occur in a variety of different foodstuffs. Depending on its speciation, Se is more or less bioavailable to human subjects. In the present study, the role of speciation as a determinant of Se bioavailability was addressed with an investigation of species-specific mechanisms of transport at the intestinal level. The present work focused on four distinct Se compounds (selenate (Se(VI)), selenite (Se(IV)), selenomethionine (SeMet) and methylselenocysteine (MeSeCys)), whose intestinal transport was mimicked through an in vitro bicameral model of enterocyte-like differentiated Caco-2 cells. Efficiency of Se absorption was shown to be species dependent (SeMet>MeSeCys>Se(VI)>Se(IV)). In the case of SeMet, MeSeCys and Se(VI), the highly polarised passage from the apical to basolateral pole indicated that a substantial fraction of transport was transcellular, whilst results for Se(IV) indicated paracellular diffusion. Passage of the organic Se species (SeMet and MeSeCys) became saturated after 3 h, but no such effect was observed for the inorganic species. In addition, SeMet and MeSeCys transport was significantly inhibited by their respective S analogues methionine and methylcysteine, which suggests a common transport system for both kinds of compounds. © 2012 The Authors.
Beguin P.,Institute Des Science Of La Vie And Uclouvain |
Schneider A.-C.,Institute Des Science Of La Vie And Uclouvain |
Mignolet E.,Institute Des Science Of La Vie And Uclouvain |
Mignolet E.,Catholic University of Louvain |
And 3 more authors.
In Vitro Cellular and Developmental Biology - Animal | Year: 2014
Human colon carcinoma cell lines such as Caco-2 cells, model of mature enterocytes and T84 cells, model of crypt cells are useful to study interactions between nutrient processing and metabolic functions at intestinal level. Our study aimed at comparing the ability of Caco-2 and T84 cells (1) to incorporate dietary polyunsaturated fatty acids (PUFA), (2) to process them and (3) to sort them into neutral lipids (NL), free fatty acids (FFA) and phospholipids (PL). Caco-2 and T84 cells were exposed to a 7-day long supplementation with PUFA. The amounts of fatty acids accumulated and incorporated into the NL, FFA or PL fractions were higher in Caco-2 than in T84 cells. Caco-2 cells were able to significantly elongate C18 PUFA and C20 PUFA of both n-3 and n-6 families. In contrast, T84 cells were unable to elongate the n-6 fatty acids whereas elongation of n-3 fatty acids was detectable but marginal. Similarly, a Δ6 desaturase activity was observed in Caco-2 but not in T84 cells. In T84 cells, each exogenous fatty acid was predominantly accumulated in the PL fraction. In Caco-2 cells, C20 fatty acids and C18:2n-6 was preferentially accumulated in the PL fraction, while C22 PUFA and C18:3n-3 was preferentially accumulated in the NL fraction. Overall, this study has shown that Caco-2 and T84 cells, as models of intestinal mucosal cells, present large differences in PUFA accumulation capacity, specific elongase and desaturase activities and distribution pattern of exogenous PUFA and of their metabolites in the lipid classes. © 2013 The Society for In Vitro Biology.
PubMed | Institute Des Science Of La Vie And Uclouvain
Type: Journal Article | Journal: Toxicology in vitro : an international journal published in association with BIBRA | Year: 2010
Enterocytes regulate gut maintenance and defence by secreting and responding to inflammatory mediators and by modulating the intestinal epithelial permeability. In order to develop an in vitro model of the acute phase of intestinal inflammation, Caco-2 cells were exposed to the inflammatory mediators IL-1beta, TNF-alpha, IFN-gamma and LPS, and the importance of several experimental parameters, i.e. cell differentiation, stimulus nature, concentration and combination on the inflammatory response was assessed by measuring the production of IL-6, IL-8, PGE-2 and NO and by evaluating the monolayer permeability. A maximal increase in IL-8, IL-6 and PGE-2 production and monolayer permeability was observed when using the cytokines simultaneously at their highest level, but this relied mainly on IL-1beta. The effects of TNF-alpha on IL-8 and IL-6 or NO production were stronger upon combination with IL-1beta or IFN-gamma, respectively, whereas cells were unaffected by the presence of LPS. Although NO production, induced by IFN-gamma-containing combinations, was observed only in differentiated cells, general inflammatory response was higher in proliferating cells. The use of a mixture of IL-1beta, TNF-alpha and IFN-gamma thus accurately mimics intestinal inflammatory processes, but cell differentiation and stimuli combination are important parameters to take into account for in vitro studies on intestinal inflammation.