A blend of botanicals reduces LPS-induced inflammation and disruptive effects on apical-out porcine enteroids

To better study host-microbial interactions or stressful conditions that affect the porcine intestinal epithelium in-vitro it is possible to use porcine apical-out enteroids, that better represent the intestinal epithelium compared to enterocytes monolayers or basal-out organoids. Botanicals are an interesting class of bioactive compounds with antioxidant and anti-inflammatory properties that help to support intestinal health. The aim of this study was to investigate the ability of a thymol-based blend of botanicals (BOT) to reduce the disruptive effects of LPS-induced inflammation on apical-out porcine enteroids. Intestinal crypts were obtained from 35 days-old pigs’ jejunum and they were used to generate apical-out enteroids. Those were then challenged with lipopolysaccharide (LPS) for 6h, in the absence or presence of BOT. FD4 paracellular permeability and gene expression were then evaluated. Apical-out porcine enteroids responded to the LPS challenge by showing a 70% increase in FD4 paracellular-permeability, a 2-fold increase in IL1β, IL8, and mucin 4 (MUC4) levels (p<0.01), and a 3-fold increase in IL6 mRNA expression (p<0.01). Moreover, a decrease in zonula occludens 1 (ZO1) and occludin (OCCL) levels was observed. BOT showed beneficial properties in reducing all the effects connected to the LPS challenge. FD4 permeability was decreased by 35% (p<0.05), and IL1β, IL6, IL8, and MUC4 expression were significantly reduced compared to the challenged control. Moreover, BOT increased the ZO1 and OCCL levels by 50% (p<0.05), acting as a barrier-reinforcing agent. BOT showed interesting anti-inflammatory properties, being able to maintain epithelial integrity thus reducing LPS-induced damage in-vitro. This thymol-based blend has the potential to be further investigated as a feed additive to improve the pigs’ ability to overcome stressful phases during their lifecycle.