Jianpi Fuzheng Xiaoji formula ameliorates gastric precancerous lesions via suppression of neutrophil extracellular trap formation

ETHNOPHARMACOLOGICAL RELEVANCE: Gastric precancerous lesions (GPL), the inflammation-driven precursors of gastric cancer (GC), may be propelled by neutrophil extracellular traps (NETs). Jianpi Fuzheng Xiaoji formula (JPFZXJ) is a standardized herbal formulation that translates the ancient "reinforce healthy qi to eliminate pathogenic factors" doctrine of the Huangdi Neijing into modern practice. It is pharmacologically optimized from the classical Han-dynasty pair Lizhong Wan and Banxia Xiexin Tang for targeted intervention against GPL and has been clinically used for its management. Nevertheless, the molecular mechanisms underlying its protective effects remain incompletely defined. AIM OF THE STUDY: To determine whether NET drives GPL and to elucidate how JPFZXJ interrupts this process. MATERIALS AND METHODS: JPFZXJ was chemically profiled by ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS). The transcriptomic dataset GSE191275 was analysed to quantify immune infiltration in non-atrophic gastritis (NAG), intestinal metaplasia (IM) and GC. In vitro, normal gastric epithelial cell-neutrophil co-cultures were employed to dissect reciprocal signalling. In vivo, a mouse GPL model induced by cyclic N-methyl-N-nitrosourea (MNU) administration was used to map neutrophil dynamics. Ablation of neutrophils or macrophages subsequently identified the principal cellular source of NET release. GPL mice were orally administered JPFZXJ for 10 weeks; infiltrating neutrophils were quantified by flow cytometry, and the capacity of JPFZXJ to attenuate GPL progression through NET inhibition was then evaluated. RESULTS: JPFZXJ significantly attenuated gastric mucosal pathological injury, ameliorated IM and dysplasia (Dys), down-regulated pro-inflammatory cytokines and chemokines, and suppressed neutrophil chemotaxis from peripheral blood to the gastric mucosa, thereby reducing neutrophil recruitment in GPL mucosa. Additionally, JPFZXJ directly inhibited NET formation in GPL mucosa. CONCLUSION: JPFZXJ alleviates GPL through dual mechanisms: curtailing neutrophil recruitment to limit NET sources and directly suppressing NET formation, underscoring its potential as an immunomodulatory approach against gastric carcinogenesis.