Objective  To analyze the chemical components of Xiaoshi Lidan pills by using UPLC-MS/MS and explore the mechanism of Xiaoshi Lidan pills in the treatment of cholelithiasis through network pharmacology and molecular docking techniques.

Methods  The pharmacologically active components of Xiaoshi Lidan pills were analyzed through UPLC-MS/MS and compared with standard references. Potential targets of these components were obtained by searching the TCMSP and ETCM databases, and disease-related targets for cholelithiasis were identified using the DisGeNET database. The overlapping targets were used to construct a protein-protein interaction (PPI) network in the String database, and a "drug-component-target"network was built using Cytoscape 3.9.1. GO and KEGG enrichment analyses were performed for the core targets. Finally, the top 5 compounds with strong activity were selected as ligands for molecular docking with the screened disease target genes. The anti-inflammatory activity was verified by RAW264.7 cells, and the mRNA expression of TNF-α and other inflammatory factors was detected by RT-PCR.

Results  UPLC-MS/MS identified 30 compounds in Xiaoshi Lidan pills, among which baicalin, quercetin, wogonin, baicalein-7-O-glucuronide, and emodin were identified as key components of Xiaoshi Lidan pills. Network pharmacology identified 107 targets associated with cholelithiasis, with Alb, TP53, ESR1, TNF, and INS identified as core targets. GO analysis indicated the involvement in inflammation response and steroid binding, while KEGG pathways were primarily related to lipid metabolism, atherosclerosis, and the TNF signaling pathway. Molecular docking analysis and anti-inflammatory screening in vitro showed that Xiaoshi Lidan pills exhibited certain anti-inflammatory activity by regulating inflammatory factors such as TNF and inhibiting NO production through baicalein, quercetin, emodin and other components.

Conclusion  Xiaoshi Lidan pills exerts its therapeutic effect on cholelithiasis by regulating TNF-related pathways through components such as baicalin, thereby inhibiting the inflammatory response.

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