Differential Effects of Oligosaccharides, Antioxidants, Amino Acids and PUFAs on Heat/Hypoxia-Induced Epithelial Injury in a Caco-2/HT-29 Co-Culture Model

Abstract

(1) Exposure of intestinal epithelial cells to heat and hypoxia causes a (heat) stress response, resulting in the breakdown of epithelial integrity. There are indications that several categories of nutritional components have beneficial effects on maintaining the intestinal epithelial integrity under stress conditions. This study evaluated the effect of nine nutritional components, including non-digestible oligosaccharides (galacto-oligosaccharides (GOS), fructo-oligosaccharides (FOS), chitosan oligosaccharides (COS)), antioxidants (α-lipoic acid (ALA), resveratrol (RES)), amino acids (l-glutamine (Glu), l-arginine (Arg)) and polyunsaturated fatty acids (PUFAs) (docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA)), on heat/hypoxia-induced epithelial injury. (2) Two human colonic cell lines, Caco-2 and HT-29, were co-cultured and pre-treated with the nutritional components for 48 h. After pre-treatment, the cells were exposed to heat/hypoxia (42 °C, 5% O2) for 2 h. Epithelial integrity was evaluated by measuring trans-epithelial electrical resistance (TEER), paracellular Lucifer Yellow (LY) permeability, and tight junction (TJ) protein expression. Heat stress and oxidative stress levels were evaluated by determining heat-shock protein-70 (HSP-70) expression and the concentration of the lipid peroxidation product malondialdehyde (MDA). (3) GOS, FOS, COS, ALA, RES, Arg, and EPA presented protective effects on epithelial damage in heat/hypoxia-exposed Caco-2/HT-29 cells by preventing the decrease in TEER, the increase in LY permeability, and/or decrease in TJ proteins zonula occludens-1 (ZO-1) and claudin-3 expression. COS, RES, and EPA demonstrated anti-oxidative stress effects by suppressing the heat/hypoxia-induced MDA production, while Arg further elevated the heat/hypoxia-induced increase in HSP-70 expression. (4) This study indicates that various nutritional components have the potential to counteract heat/hypoxia-induced intestinal injury and might be interesting candidates for future in vivo studies and clinical trials in gastrointestinal disorders related to heat stress and hypoxia.

Effect of Digested Selected Food Items on Markers of Oxidative Stress and Inflammation in a Caco-2-Based Human Gut Epithelial Model

“…Whichever the source, MDA can be involved in intracellular adduct formation, such as with cellular DNA, proteins, as well as membranes [52]. MDA production by a Caco-2 co-culture cell model was shown to be reduced by antioxidants such as resveratrol and anti-inflammatory compounds such as eicosapentaenoic acid [53], though such further reductions with antioxidants were not observed in the present study, again possibly indicative of the MDA origin from the food matrix itself.…”

Cellular Uptake and Transport Mechanism Investigations of PEGylated Niosomes for Improving the Oral Delivery of Thymopentin

“…It can be seen that Caco-2 and HT-29 cells with ratios of 1:0, 7:1, and 9:1 met the requirement of tissue integrity. Because of the benefits of the co-culture of Caco-2 and HT-29 cells, it provides the presence of both absorptive and goblet cells, both of which have different culture requirements for optimal growth and function to mimic intestine tissue in vivo [ 38 , 39 , 40 , 41 , 42 ]. Therefore, the ratio of 9:1 was selected based on the presence of both cell types and highest TEER value for subsequent uptake and transport studies.…”

Wild blueberry (V. angustifolium) improves TNFα-induced cell barrier permeability through claudin-1 and oxidative stress modulation in Caco-2 cells

“…Moreover, urolithin-A increased the expression of heme oxygenase-1, an Nrf2-dependent gene with a crucial role in the protection against oxidative stress and inflammation. Conversely, Lian and colleagues 50 evaluated the effect of resveratrol, on a co-culture model of Caco-2 and HT29 in a condition of hypoxia and heat-induced epithelial injury. The authors found that the exposure to heat and hypoxia degraded the barrier integrity, as observed by the reduction of TEER values and the expression of zo-1 and claudin-3 proteins.…”