Xiangsha Liujunzi Tang: A Promising Traditional Chinese Herbal Formula for Healing Gastric Mucosal Injury Through Pyroptosis Pathway Modulation

Gastric mucosal injury is a common gastrointestinal disorder characterized by a high recurrence rate, and current clinical treatments remain limited due to adverse reactions. Xiangsha Liujunzi Tang (XSLJZT) is a classic traditional Chinese medicine formula with well-documented gastroprotective efficacy in clinical practice; however, its underlying pharmacological mechanism, particularly its regulatory role in pyroptosis, remains poorly elucidated. In this study, bioinformatics and molecular docking were employed to identify targets and pathways associated with gastric mucosal injury. XSLJZT was manufactured and quality-controlled using the liquid chromatography-tandem mass spectrometry (LC-MS/MS). Hematoxylin-eosin (H&E) staining along with transmission electron microscopy was utilized to analyze the morphology and ultrastructure of the gastric mucosal surface. Periodic acid-Schiff (PAS) staining and ELISA were utilized to measure gastric mucosal glycoproteins and the release of pro-inflammatory and anti-inflammatory cytokines in rats with IND-induced gastric mucosal injury. Immunofluorescence was employed to detect the expression levels of GSDMD-N, p-NF-κB, p-IKKβ, NLRP3, ASC, and cleaved-caspase-1 proteins to elucidate the potential molecular mechanisms underlying XSLJZT's gastroprotective effects. Bioinformatics analysis and molecular docking simulation indicated that key molecules of the NLRP3 inflammasome-mediated pyroptosis pathway are closely associated with gastric mucosal injury, and the active components of XSLJZT show potential binding interactions with these target proteins. Animal experiments demonstrated that XSLJZT significantly reduced the ulcer index and improved swelling, hemorrhage, and ulcers of the gastric mucosa. Additionally, XSLJZT administration significantly increased the levels of protective cytokines (IL-4, IL-10, and IL-12) while decreasing the levels of pro-inflammatory mediators (IL-18, IL-1β, and TNF-α). Further in vivo validation confirmed that XSLJZT markedly downregulated the protein expression of GSDMD-N, p-NF-κB, p-IKKβ, NLRP3, ASC, and cleaved caspase-1. These findings suggest that XSLJZT may exert a therapeutic effect on digestive disorders associated with gastric mucosal injury by modulating the NLRP3 inflammasome-mediated pyroptosis signaling pathway.