Yanbin Zhu ¹ Sijia Lu ² Xiaojuan Zhang ³ Yangji Cidan ¹ Wangdui Basang ¹ Kun Li ^2*

• ¹ Tibet Academy of Agricultural and Animal Husbandry Sciences, Lhasa, China
• ² Nanjing Agricultural University, Nanjing, China
• ³ Agriculture and Animal husbandry science and technology service station in Seni district, Naqu, China

Chinese herbal medicines are relatively cheap and have low side effects that can be effectively utilized to improve the health and to treat diseases, hence received more attention in recent years. The weaning period is a critical stage in the life of yaks especially in inducing stress in calves. Weaning stress and dietary changes can lead to a decline in physical fitness and immune system, increasing susceptibility to diarrhea and resulting in a high mortality rate of yaks during this period. Therefore, our study aimed to address this issue by adding traditional chinese medicines (TCM) formulas to the diet of yaks during weaning period. Following a dialectical analysis, three TCM formulas, mainly composed of Paeonia lactiflora, Coptis chinensis, and Dandelion were identified for their anti-inflammatory, antioxidant, and immune enhancing potentials. We explored the possible molecular mechanisms of these TCM formulas using network pharmacology analysis and investigated their effects on yaks' physiology through metabolomics. Network pharmacology analysis revealed several key targets proteins in the protein-protein interaction (PPI) network between three formulas and immune related genes, including PIK3R1, PIK3CA, JAK2, PTK2 and PYPN11.The key target proteins in the PPI network associated with metabolism-related genes included ENPP1, CYP1A1, PTGS1, members of the CYP1 family, and EPHX2. GO analysis of co-targets revealed highly enriched pathways such as protein phosphorylation, plasma membrane, and one-carbon metabolic processes. Metabolomics revealed significant changes in the abundance of metabolites including dimethyl sulfoxide, tyrphostin A25, thromboxane A2, in the intestines of weaned yaks supplemented with these chinese herbal compounds. Significant changes were also observed in pathways such as vitamin A metabolism, chloroalkane and chloroalkene degradation. Based on these findings, it can be inferred that TCM formulas improve the physical fitness of weaned yaks by enhancing antioxidant capacity, boosting immunity, and reducing intestinal inflammation. This study preliminarily elucidates the pharmacological mechanisms by which TCM formulas prevent diarrhea and improve physical fitness in weaned yaks through metabolomics and network pharmacology, paving the way for further evaluation for the effectiveness of these three formulas.