Abstract

Background : Gastric ulcer (GU) remains clinically challenging due to high recurrence rates and adverse effects of conventional drugs. The classic traditional Chinese medicine (TCM) formula Wuji Wan (WJW) exhibited empirical efficacy in the treatment of GU. However, the precise mechanisms underlying the therapeutic effects of WJW’s active ingredients remain unclear. Purpose and Methods : We developed a defined combination of eight bioactive WJW constituents (ComB) and investigated its anti-ulcer potential. In vitro , the cytoprotective effects of ComB were assessed in ethanol-injured GES-1 gastric epithelial cells. In vivo , rats with ethanol-induced GU were treated with ComB to evaluate mucosal repair. Mechanistic studies based on proteomics, target interaction studies, and cascade analysis of signaling pathways were conducted to dissect the mode of action of ComB. Results : ComB showed significant protective effects, enhanced the viability of ethanol-injured GES-1 gastric epithelial cells, and reduced mucosal damage in the rat model of GU. Mechanistic studies revealed that ComB can directly bind to both STAT3 and PKR, effectively disrupting their molecular interaction and blocking the nuclear translocation of STAT3. This targeted intervention promoted autophagic flux and substantially alleviated endoplasmic reticulum stress (ERS) through coordinated regulation of the STAT3/PKR-autophagy signaling axis. Conclusions : By directly disrupting the STAT3-PKR interaction, ComB inhibited STAT3 nuclear translocation, activated autophagic flux, and alleviated ERS, thereby protecting gastric mucosa against ethanol-induced damage. This study identified ComB as a promising traditional Chinese medicine combination for ethanol-induced GU therapy by inhibiting ERS through targeting STAT3/PKR to promote autophagy signaling.