Effect of Tongxie Yaofang formula on the gut microbiota of diarrhea model mice: a combined strategy of network pharmacology, molecular docking, and 16S rRNA gene sequencing

Tongxie Yaofang (TXYF) is a classic formula for liver stagnation and spleen deficiency diarrhea, but its mechanisms involving the gut microbiota remain unclear. This study aimed to elucidate the mechanisms TXYF by integrating network pharmacology, molecular docking, and 16S rRNA gene sequencing. We employed network pharmacology and molecular docking to identify bioactive compounds, targets, and their associations with gut microbiota metabolites. Sequencing of the 16S rRNA gene revealed changes in the gut microbiota in a diarrheal mouse model after TXYF intervention. Thirty-nine active compounds (525 targets) were identified. Intersection analysis yielded 44 shared targets among TXYF, diarrhea, and gut microbiota metabolites. Molecular docking demonstrated favorable binding of the selected active compounds and representative gut microbiota-derived metabolites with the core targets GSK3B, TP53, and MAPK3. Sequencing of the 16S rRNA gene also showed that TXYF significantly modulated gut microbiota structure, notably altering abundances of Corynebacterium stationis, Proteus vulgaris, and Proteus mirabilis. TXYF treats diarrhea by modulating gut microbiota composition and maintaining microbial homeostasis via a multi-component, multi-target mechanism, supporting its potential as a microbiota-targeted therapy.