Regionally specific dysregulation of gut microbiota and host–metabolite interface in podophyllotoxin-induced enterotoxicity based on the toxicological evidence chain concept

Podophyllotoxin (PPT) induces enterotoxicity through gut microbiota dysbiosis; however, the influence of intestinal regional specificity on toxicity remains unclear. This study investigated segment-specific alterations in PPT-induced enterotoxicity using a toxicological evidence chain framework. Male Sprague-Dawley rats received oral PPT (10, 15, or 20 mg/kg) daily for 4 days. Intestinal toxicity was evaluated using histopathology and quantitative real-time polymerase chain reaction analysis of tight junction proteins and pro-inflammatory cytokines. 16S rRNA sequencing and metabolomic profiling were performed on the contents of the small intestine, cecum, and colon. Microbe-metabolite interactions were analyzed using Spearman's correlation and K-means clustering. The results demonstrated that PPT exposure induced significant enterotoxicity, as evidenced by diarrhea, villus damage, goblet cell loss, barrier impairment, and inflammation. 16S rRNA sequencing revealed a consistent depletion of beneficial genera Romboutsia , Lombactobacillus , and Turicibacter, alongside an expansion of pathogenic Escherichia-Shigella across all gut segments. Metabolomic analysis revealed profound disturbances in key metabolic pathways. Notably, a putative co-regulated protective axis encompassing vitamin B6, histidine, and tryptophan metabolism was altered, with central metabolites such as 4-pyridoxic acid and β-alanyl-L-histidine being significantly downregulated. Critically, these microbial and metabolic alterations exhibited distinct spatial patterns. Network analysis indicated that the correlations between depleted beneficial bacteria and downregulated protective metabolites were stronger in the cecum and colon than in the small intestine. Collectively, these findings provide systematic evidence that PPT-induced enterotoxicity is associated with region-specific disturbances in the gut microbiota–metabolite network, which may serve as a potential target for mitigating PPT-induced intestinal damage. • Podophyllotoxin (PPT) causes severe intestinal injury with impaired barrier function and inflammation. • PPT disrupts gut microbiota by decreasing beneficial genera and increasing pathogenic bacteria across intestinal segments. • PPT-induced enterotoxicity is mediated by region-specific microbiota-metabolite disorders, with the strongest correlation in the cecum and colon.