Gut Microbiota-derived Metabolites Contribute Negatively to Hindgut Barrier Function Development at the Early Weaning Goat Model

Early weaning induces intestinal injury, leading to a series of long-term symptoms such as inflammation, malabsorption and diarrhea. In this study, we hypothesized that microbes and their metabolites modulate the host's inflammatory response to early weaning stress in a goat model. A total of 18 female Tibetan goat kids ( = 9) were weaned from their mothers at 28 d (D28) and 60 d (D60) postpartum. D60 and D28 groups were fed the same solid diet ad libitum from weaning to 75 d of age. The colonic epithelium was subject to RNA-sequencing, the caecal digesta metabolomics were assessed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the caecal microbiota composition was analysed by 16S ribosomal RNA gene sequencing. We found that early weaning substantially increased the colonic pro-apoptotic gene expression of B-cell lymphoma associated X (), caspase-9, and caspase-3, and decreased the expression of zonula occludens-1 () and claudin-1 ( < 0.01). In addition, a significant enrichment was observed in the hindgut of early-weaned goats ( < 0.01), which negatively correlated with lysophosphatidylcholine products. Similarly, the chemokine signaling, signaling, and peroxisome proliferators-activated receptor () signaling pathways were upregulated in the colonic mucosa of the early-weaned goats. By applying caecal microbiota transplantation from goats to defaunated C57/6J mice, we confirmed that caecal microbiota of D28 goat kids increased the relative abundance of and significantly up-regulated the genes of , G protein-coupled receptor () , , fatty acid binding protein 6, nuclear receptor subfamily 1 group H member 3, angiotensin converting enzyme 2, and expression ( < 0.05), and decreased , and claudin-1 protein expression in the mice jejunum and colon ( < 0.001). These results proposed that the hindgut microbiota and metabolites mediate the barrier function weakening during early weaning, and the relative abundance of was negatively correlated with the hindgut barrier gene expression. This study demonstrates how weaning stress can affect key host-microbe interaction regulators in the hindgut, in a lysophosphatidylcholine dependent and independent manner. Furthermore, based on our mice data, these results are transferable to other mammal species.