Tengchuan compound mixture ameliorates Mycoplasma Synoviae-induced synovitis: An integrated network pharmacology, metabolomics, and 16S rRNA sequencing study

Mycoplasma synoviae (MS) is a prevalent pathogen in poultry farming, often causing infectious synovitis and arthritis in poultry, resulting in joint swelling, lameness, and growth retardation, thereby significantly impacting production efficiency. Tengchuan compound mixture (TCM), a traditional Chinese medicine formulation designed to against MS infection effectively, but its active components and mechanisms isn't elucidated. This study utilized liquid chromatography-mass spectrometry (LC-MS) to identify the active components in TCM and employed network pharmacology to predict key targets and pathways in against MS infection. A model of MS infection was established in specific-pathogen-free (SPF) chickens to assess the pharmacodynamic effects of TCM on parameters such as body weight, clinical manifestations, histopathology of tarsal joints, and serum inflammatory markers. Metabolomics analysis and 16S rDNA sequencing were conducted to elucidate the potential mechanism of TCM in preventing and treating MS infection. Western blot analysis confirmed the core targets identified by network pharmacology. TCM treatment effectively mitigated MS-induced joint lesions by suppressing the phosphorylation of proteins associated with the JAK2-STAT3 and PI3K-AKT signaling pathways. Pharmacokinetic analysis identified 18 major potential active ingredients. Network pharmacology identified 147 common targets of TCM against MS infection, including 13 core targets such as SRC, JAK2, PIK3CA, and STAT3. Metabolomics analysis revealed that MS infection disrupted amino acid and energy metabolism in chickens, which was partially restored by TCM treatment. Additionally, 16S rDNA sequencing indicated that TCM could recovery the gut microbiota disrupted by MS infection. In conclusion, this integrated multi-omics study provides mechanistic insights into TCM's effects during MS infection, highlighting its role in ameliorating infection-induced inflammation and joint pathology while modulating host metabolic pathways and gut microbiota. These findings offer a comprehensive understanding of TCM's therapeutic potential for managing Mycoplasma synoviae infection.