Dissection of potential mechanism of Bao-Yuan tablet against Spleen-Qi Deficiency by UPLC-Q/TOF MS and network pharmacology.

Bao-Yuan tablet (BYT), a modern formulation derived from the classic Bao-Yuan decoction (BYD) first documented in the Ming Dynasty of China, is used as a therapeutic agent for Spleen-Qi Deficiency (SQD). However, its pharmacodynamic effects and underlying mechanisms remain unclear. This study aimed to investigate its therapeutic effects and mechanism against SQD induced by a multifactorial method (cold-purging, fatigue, and irregular diet). BYT treatment significantly improved behavioral symptoms, increased thymus index, reduced serum IgA/IgG levels, elevated D-xylose content, and suppressed TNF-α levels in SQD mice. A total of 141 compounds were identified in BYT by UPLC-Q/TOF MS, including five previously undescribed compounds. Based on these identified components, network pharmacology predicted that TNF, IL-1β, and TLR4 are core targets of BYT against SQD, involving Toll-like receptor and TNF signaling pathways. Western blot validation confirmed that BYT dynamically regulated the splenic TLR4/NF-κB pathway: suppressing TLR4 and p65 phosphorylation at day 7, while restoring TLR4 and reducing elevated total p65 at day 14. This study provides the first experimental evidence for BYT against SQD and integrates chemical profiling, network prediction, and experimental validation of its multi-target mechanism, providing a scientific basis for further quality control and clinical application of BYT.