An integrative analysis combining network pharmacology, transcriptomics, and experimental validation of the Tibetan medicine Renqing Changjue in gastric ulcer protection.

ETHNOPHARMACOLOGICAL RELEVANCE: Renqing Changjue (RQCJ), a time-honored Tibetan prescription, has long been used to treat "heat toxin"-related disorders, particularly those affecting the digestive system. Despite its extensive clinical use, the pharmacodynamic substances and molecular mechanisms underlying its anti-ulcer effects remain poorly defined.

AIM OF THE STUDY: This study aimed to identify the key active constituents, core targets, and underlying mechanisms by which RQCJ protects against ethanol-induced gastric ulcer.

MATERIALS AND METHODS: An integrative multi-omics framework combining UPLC-QTOF-MS analysis, network pharmacology, transcriptomics, molecular docking, and in vivo assays was established. Ethanol-induced gastric ulcer mice and ethanol-injured GES-1 cells were used to evaluate histopathological recovery, oxidative stress and inflammatory markers, apoptosis-related proteins, and PI3K/AKT signaling activity.

RESULTS: UPLC-QTOF-MS analysis annotated 27 major constituents in RQCJ. Network and transcriptome analyses identified 110 overlapping targets, with enrichment centered on the PI3K/AKT pathway. In addition to protecting GES-1 cells, RQCJ administration markedly attenuated gastric mucosal lesions, alleviated oxidative injury, suppressed pro-inflammatory cytokine release, and inhibited apoptosis by modulating the Bax/Bcl-2 balance and the caspase cascade. Western blotting and immunohistochemistry further confirmed phosphorylation-driven activation of the PI3K/AKT signaling axis.

CONCLUSION: RQCJ exerts potent gastroprotective effects through multi-target modulation of oxidative stress, inflammation, and apoptosis, primarily by activating the PI3K/AKT pathway. These findings provide mechanistic insight into the traditional Tibetan use of RQCJ and establish a scientific foundation for its development as a natural therapeutic for gastric ulcer.