Atractylodis Macrocephalae Rhizoma ameliorates diarrhea induced by cold drinks and a high-fat diet by remodeling gut microecology and restoring barrier function.

Background: Atractylodis Macrocephalae Rhizoma (AMR) has traditionally been utilized for treating spleen deficiency diarrhea. Nevertheless, the effects and mechanisms of AMR on diarrhea caused by the consumption of cold drinks and a high-fat diet (CDHFD) remain insufficiently understood.

Aim: This study aimed to explore the therapeutic effects and mechanisms of AMR in treating CDHFD-induced diarrhea.

Methods: AMR was prepared as an aqueous extract, and its chemical composition was analyzed using UPLC-ESI-MS. A diarrhea model was established in ICR mice by exposure to CDHFD for four weeks, with AMR (low/high doses) administered concurrently via oral gavage. Bowel movements were evaluated using indicators such as fecal water content. Systemic inflammation was assessed by measuring pro-inflammatory cytokines via ELISA and performing peripheral blood cell counts. Intestinal barrier integrity was examined via H&E, AB-PAS staining, and immunofluorescence of tight junction proteins. Gut microbiota profiling was performed using 16S rDNA sequencing. Serum lipopolysaccharide (LPS) levels were measured via ELISA to assess translocation. Finally, the regulatory effects of AMR on the A20/TRAF6/NF-κB signaling pathway were validated using Western blotting. Spearman's correlation analysis was employed to integrate microbiota changes with host inflammatory phenotypes.

Results: AMR significantly ameliorated CDHFD-induced diarrhea. Mechanistically, AMR remodeled the gut microecology by enriching beneficial bacteria, particularly Lachnospiraceae_NK4A136 and norank_f_Muribaculaceae. Concurrently, it increased the number of goblet cells and regulated the expression of tight junction proteins to repair intestinal barrier damage and reverse hyperpermeability. The restoration of barrier function effectively blocked the systemic translocation of LPS, which subsequently inhibited the hyperactivation of the NF-κB signaling pathway, thereby reducing systemic inflammation and ultimately alleviating diarrhea.

Conclusion: AMR exerts protective effects against CDHFD-induced diarrhea through microbiota-driven intestinal barrier restoration, which sequentially blocks the activation of the LPS/NF-κB inflammatory pathway.