The Anti-Inflammatory Action of Artichoke, Fenugreek and Caigua (AFC) Original Blend in an Inflammatory Bowel Disease In Vitro Model ^†

Abstract

Background and objectives: The incidence of chronic inflammatory pathologies has incrementally increased in recent years, as in the case of inflammatory bowel disease (IBD), which is characterized by intestinal epithelial barrier disruption, increased inflammatory mediator production and excessive tissue injury. Changes in eating habits might have played a key role in this scenario. Therefore, the interest in specific diet development and in functional food formulation has been growing. Phytoextracts from several origins, from plants to waste, enriched in bioactive molecules, alone or combined, might be a resource for the obtainment of an efficient synergistic beneficial. Thus, the aim of this study consists of evaluating the protective effects of artichoke, fenugreek and caigua (AFC) phytoextract original blend. Methods: In order to mimic the intestinal barrier’s inflammatory environment, Caco-2 cells were cultured and polarized on a transwell system and then exposed to a pro-inflammatory cytokine cocktail (TNFα and IL-1β). Before being exposed to an inflammatory stimulus, cells were pre-treated with an AFC digested blend, according to the INFOGEST in vitro static digestion protocol. After digestion, the content of active substances within the blended extract (ACFB) was revealed by UHPLC–ESI–HRMS analysis. The AFC digested extract’s protective effect was evaluated by measuring the transepithelial resistance (TEER) as a marker of barrier integrity and analysing the nuclear factor kappa B (NF-κB) pathway. Results: The TEER values improved in cells which were pre-treated with the AFC blend, relative to inflamed cells, suggesting a regulation in tight junction protein expression and/or localization. The transcription factor p65NF-κB is activated by phosphorylation under cytokine exposure, with a 160% increase in its target COX-2. Moreover, a 40-fold increase in IL-8 release was observed. Interestingly, in cells pre-treated with the AFC blend, the activated p65NF-κB was halved, compared to inflamed cells only. Furthermore, a consequent reduction by about 50% for COX-2 and by 30% for IL-8 was observed. Discussion: Taken together, these results highlight the anti-inflammatory potential of the AFC blend, probably due to the presence of flavonoids such as luteolin, apigenin and chrysin. This experimental evidence suggests that an AFC blend could be a good ingredient for food functionalization if further used in nutritional strategies.