Abstract

Background Chronic atrophic gastritis (CAG) is a pivotal intermediate stage in the progression from chronic gastritis to gastric cancer (GC), and it poses a serious threat to human health. Astragaloside IV (AS-IV), a major bioactive compound derived from the traditional Chinese medicinal herb Astragalus membranaceus , has demonstrated therapeutic potential against both CAG and GC. Nevertheless, the exact biological mechanisms underlying these effects remain unclear. Purpose This research was conducted to assess the effects of AS-IV on gastric inflammation-cancer transformation and to elucidate its underlying mechanisms. Methods We collected gastric tissue and blood samples from healthy individuals, patients with CAG, and patients with GC. The levels of CXCL5 and CXCR2 were quantified using ELISA and immunohistochemistry. To establish in vitro models of CAG and GC, we employed the GES-1 and MKN45 cell lines. Experimental interventions included lentiviral-mediated CXCL5 knockdown (shCXCL5-KD), CXCR2 knockdown (shCXCR2-KD), CXCL5 overexpression (shCXCL5-OE), and treatment with AS-IV. Cellular responses were evaluated using multiple assays: Western blotting to assess protein expression, quantitative real-time polymerase chain reaction (RT-qPCR) to measure mRNA levels, CCK-8 for cell viability, ELISA for cytokine quantification, and wound healing, along with Transwell assays to assess cell migration. For in vivo validation, CAG models were established in Sprague–Dawley (SD) rats and GC models in BALB/c nude mice, with AS-IV administered according to the designated experimental groups. Systematic evaluations were performed using multiple techniques: hematoxylin and eosin (H&E) staining to assess histopathological changes, immunohistochemistry to determine protein localization, Western blotting to analyze signaling pathways, and ELISA to quantify systemic cytokine levels. Together, these approaches enabled a comprehensive, multi-level analysis of the therapeutic mech