In this study, 10 mg/kg of loperamide was administered to induce a constipation mouse model, aimed at investigating the protective effects of lacto-N-neotetraose on constipation and its impact on the intestinal flora in mice.Six-week-old Balb/c mice were randomly assigned to three groups, including the normal control group (Ctrl), the model group (Model), and the lacto-N-neotetraose treatment group (LNnT).Mice were fed the respective treatments for 14 days.The mice were analyzed for cecal water content, gastric emptying rate, small intestinal transit rate, time to the first black stool, levels of regulatory peptides in the gastrointestinal tract, intestinal histology, cytokine levels, MUC2 secretion, and intestinal flora.Results indicated that compared to the Model group, the fecal water content, gastric emptying rate, and small intestinal transit rate in lacto-N-neotetraose-treated mice increased by 20.89%, 19.94%, and 74.48%, respectively.The time to the first black stool appearance was reduced by 91 minutes.Gastrin, gastric motility, and substance P levels increased by 28.76%, 45.58%, and 17.06%, respectively, while vasoactive intestinal peptide content decreased by 27.60%.Additionally, prokinetic peptide and prokaryotic peptide levels both decreased by 27.60%.The levels of pro-inflammatory cytokines (IL-1 and IL-1β) were significantly reduced, while the levels of the anti-inflammatory cytokine IL-6 were significantly increased.In addition, lacto-N-neotetraose enhanced MUC2 gene expression and protein levels.Furthermore, lacto-N-neotetraose treatment enhanced the relative abundance of Ackermannia, Prevotella anomalosa, and Bifidobacterium, while reducing the relative abundance of Eisenbergia, Lachnoclostridium, and Ruminalococcus.In conclusion, lacto-N-neotetraose alleviated constipation symptoms, intestinal inflammation, and mucin secretion induced by loperamide in mice.The underlying mechanism may involve the regulation of intestinal flora and the promotion of beneficial bacterial growth.
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