Eugenol (4-allyl-2-methoxyphenol) is an essential oil component, possessing anti-microbial, anti-inflammatory and anti-oxidative properties, however the effect of eugenol on porcine gut inflammation has not yet been investigated. In this study, an in vitro lipopolysaccharide (LPS)-induced inflammation model in porcine intestinal epithelial cells (IPEC-J2) has been set up. Cells were pre-treated with 100 μM (16.42 mg/L) eugenol for 2 h followed by 10 μg/mL LPS stimulation for 6 h. Pro-inflammatory cytokine secretion, reactive oxygen species, gene expression of pro-inflammatory cytokines, tight junction proteins and nutrient transporters, the expression and distribution of zonula occludens-1 (ZO-1), trans-epithelial electrical resistance (TEER) and cell permeability were measured to investigate the effect of eugenol on inflammatory responses and gut barrier function. The results showed that eugenol pre-treatment significantly suppressed the LPS-stimulated interleukin 8 (IL-8) level and the mRNA abundance of tumor necrosis factor α (TNF-α); restored the LPS-stimulated decrease of mRNA abundance of tight junction proteins zonula occludens-1 (ZO-1), occludin (OCLN) and the mRNA abundance of nutrient transporters B0-system neutral amino acid co-transporter (B0AT1), system ASC sodium-dependent neutral amino acid exchanger 2 (ASCT2), apical sodium-dependent glucose transporter 1 (SGLT1), excitatory amino acid transporter 1 (EAAC1) and peptide transporter 1 (PepT1). In addition, eugenol improved the expression and even redistribution of ZO-1 and tended to increase TEER value and maintained the barrier integrity. In conclusion, a low dose of eugenol attenuated inflammatory responses and enhanced selectively permeable barrier function during LPS-induced inflammation in the IPE-J2 cell line.
Essential oils (EO) are defined as plant-derived natural bioactive compounds, which can have positive effects on animal growth and health due to their antimicrobial and antioxidative properties. However, EO are volatile, can evaporate quickly, and be rapidly absorbed in the upper gastrointestinal tract. Also, due to their labile nature, the stability of EO during feed processing is often questionable, leading to variations in the final concentration in feed. Encapsulation has become one of the most popular methods of stabilizing EO during feed processing, storage, and delivery into the lower gut. The objectives of the present study were to 1) evaluate the stability of thymol microencapsulated in combination with organic acids in commercially available lipid matrix microparticles during the feed pelleting process and storage; 2) validate and demonstrate the slow release of thymol from the lipid matrix microparticles in a simulated pig gastric fluid (SGF) and a simulated pig intestinal fluid (SIF); and 3) evaluate in vivo release of thymol from the lipid matrix microparticles along the pig gut. The results showed that thymol concentration was not significantly different in the mash and pelleted feeds (P > 0.05). In the in vitro study, 26.04% thymol was released in SGF, and the rest of the thymol was progressively released in SIF until completion, which was achieved by 24 h. The in vivo study showed that 15.5% of thymol was released in the stomach, and 41.85% of thymol was delivered in the mid-jejunum section. Only 2.21% of thymol was recovered in feces. In conclusion, the lipid matrix microparticles were able to maintain the stability of thymol during a feed pelleting process and storage and allow a slow and progressive intestinal release of thymol in weaned pigs.
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