Identification of bioactive components in Wuji Wan and their protective effects against gastric ulcer via regulating PIK3CA-mediated PI3K/AKT pathway.

The classical Chinese formula Wuji Wan (WJW), which integrates Coptis chinensis, Evodia rutaecarpa, and Paeonia lactiflora, offers therapeutic benefits for gastric ulcer, as validated by its historical use in traditional practice.This study aimed to delineate the protective properties of WJW against gastric ulcer (GU) by systematically identifying its target-effective bioactive compounds and characterizing their fundamental molecular mechanisms. An integrated strategy combining UPLC-MS/MS, network pharmacology, serum metabolomics, and molecular docking was employed to identify candidate components and key targets. The binding affinity between components and the core target was validated using bio-layer interferometry (BLI), drug affinity responsive target stability (DARTS), and cellular thermal shift assay (CETSA). Via Western blotting, RT-qPCR, and immunohistochemical staining, the therapeutic effects and mechanisms were confirmed using ethanol-induced GES-1 cells and a rat GU model. Seven core bioactive components, including palmatine, coptisine, rutaecarpine, berberine, paeoniflorin, albiflorin, and evodiamine, were identified as target-effective components of WJW. The efficacy of these components against GU was mediated by targeting PIK3CA, which in turn suppressed the PI3K/AKT signaling pathway, finally suppressing inflammatory responses and inhibiting cellular apoptosis. The significance of this study lies not only in clarifying the material basis and mechanism of WJW against GU but also in showcasing a target-driven strategy for the discovery of active ingredients from complex formulae. Such an approach provides a scientific framework for developing standardized botanical drugs.