OBJECTIVE To investigate the mechanism of action of Ershiwuwei Datang pills in the treatment of gastric ulcer via network pharmacology combined with in vivo animal experiments.

METHODS Initially, ultra-high-performance liquid chromatography-linear ion trap-Orbitrap mass spectrometry(UPLC-LTQ-Orbitrap-MS) was employed to identify the blood-entering constituents of the Ershiwuwei Datang pills. Subsequently, network pharmacology techniques were employed to predict potential targets and pathways for the treatment of gastric ulcers by the Ershiwuwei Datang pills, and the results were validated through molecular docking. Finally, a rat model of gastric ulcer complicated by spleen-stomach deficiency cold syndrome was established using a combination of purgative and cold-inducing herbs, irregular feeding schedules, and administration of absolute ethanol and aspirin solution. Histopathological changes in gastric tissues were assessed via hematoxylin and eosin(HE) staining. Serum levels of TNF-α, IL-6, and GAS were measured using enzyme-linked immunosorbent assay(ELISA). Moreover, the expression levels of IL-6, Caspase-3, PI3K, and EGFR in gastric tissues were determined by quantitative real-time polymerase chain reaction(qRT-PCR) and Western blotting analysis.

RESULTS A total of 10 blood-entering constituents of Ershiwuwei Datang pills were identified. Network pharmacology analysis acquired 503 compound targets and 5856 disease-related targets. Protein-protein interaction(PPI) analysis screened out 7 core targets including TP53, SRC, PIK3CA and others. KEGG analysis indicated that signaling pathways such as PI3K and EGFR tyrosine kinase inhibitor resistance wexert key effects in the anti-gastric ulcer process of Ershiwuwei Datang pills. Furthermore, Molecular docking results showed that cosmosiin, zingerone and afzelin have strong binding capacity with the core targets. Animal experimental results showed that compared with the model group, the serum levels of TNF-α and IL-6 n each administration group were significantly reduced(P<0.01), while the levels of GAS increased; the expression levels of IL-6, Caspase-3 and PI3K in gastric tissues was reduced, and the expression level of EGFR was increased.

CONCLUSION The Ershiwuwei Datang pills exert its therapeutic effect on gastric ulcer by regulating the PI3K signaling pathway, thereby alleviating inflammatory responses and reducing cell apoptosis.