Arjunolic acid protects the intestinal epithelial barrier, ameliorating Crohn's disease-like colitis by restoring gut microbiota composition and inactivating TLR4 signalling.

BACKGROUND AND AIMS: Crohn's disease (CD) is characterized by an overabundance of epithelial cell death and an imbalance in microflora, both of which contribute to the dysfunction of the intestinal barrier. Arjunolic acid (AA) has anti-apoptotic effects and regulates microbiota efficacy. The objective of this study was to assess the impact of the treatment on colitis resembling Crohn's disease, along with exploring the potential underlying mechanism. METHODS: CD animal models were created using Il-10-/- mice, and the impact of AA on colitis in mice was evaluated through disease activity index, weight fluctuations, pathological examination, and assessment of intestinal barrier function. To clarify the direct role of AA on intestinal epithelial cell apoptosis, organoids were induced by LPS, and TUNEL staining was performed. To investigate the potential mechanisms of AA in protecting the intestinal barrier, various methods including bioinformatics analysis and FMT experiments were employed. RESULTS: The treatment for AA enhanced the condition of colitis and the function of the intestinal barrier in Il-10-/- mice. This was demonstrated by the amelioration of weight loss, reduction in tissue inflammation score, and improvement in intestinal permeability. Moreover, AA suppressed the apoptosis of intestinal epithelial cells in Il-10-/- mice and LPS-induced colon organoids, while also reducing the levels of Bax and C-caspase-3. In terms of mechanism, AA suppressed the activation of TLR4 signaling in Il-10-/- mice and colon organoids induced by LPS. In addition, AA increased the abundance of short-chain fatty acid-producing bacteria in the stool of Il-10-/- mice, and transplantation of feces from AA-treated mice improved CD-like colitis. CONCLUSIONS: The results of our study demonstrate that AA has a protective effect on the intestinal barrier in Crohn's disease-like colitis by preventing apoptosis. Additionally, this groundbreaking study reveals the capacity of AA to hinder TLR4 signaling and alter the makeup of the intestinal microbiome. The findings present fresh possibilities for treating individuals diagnosed with Crohn's disease. AA offers a hopeful novel strategy for managing Crohn's disease by obstructing crucial pathways implicated in intestinal inflammation and enhancing the gut microbiota.

Effects of citrus pulp on the composition and diversity of broiler cecal microbes.

Diet may affect gut microbial composition and diversity. There were 3 dietary groups: 0% citrus pulp diet (C), 1.5% citrus pulp diet (I), and 2.5% citrus pulp diet (II). A total of 180 healthy AA broilers (21-day old) were divided into 3 groups (C, I, and II), each group was set up with 6 replicates, and each replicate including 10 broilers (half male and female). At 42 d, the cecal contents of 18 broiler chickens were collected after slaughter. The cecal contents were analyzed using 16S rRNA sequencing technology. Compared with group C, the abundance of Firmicutes in groups I and II decreased, while the relative abundances of Bacteroidetes, Verrucomicrobia, Lactobacillus, and Faecalibacterium increased. LEfSe analysis showed that Actinobacteria, Coriobacteriia, Coriobacteriales, and Ruminococcaceae_bacterium_Marseille_P2935 in group I were significantly higher than those in group C. Bacteria, Coriobacteriales, Coriobacteriia, Coriobacteriaceae, Slackia, Bacteroides_sp_Marseille_P3132, and Lactobacillus_pontis in group II were significantly higher than those in group C. The Staphylococcaceae, Bacteroides_sp_Marseille_P3132, Macroccus, Lactobacillus_pontis, and Streptococcus_equinus in group II were significantly higher than those in group I. Functional predictions indicated that the cecal microbiota of broilers fed the 2.5% citrus pulp diet was more tend to utilize carbohydrates through glycolytic/gluconeogenesis metabolism. Adding citrus pulp to the diet affects the microbial composition and has important implications for studying gut health and improving economic benefits.