Effects of Pyloric Ligation-Induced Gastric Injury on Glomerular Mesangial Cells and the Intervention Mechanism of Xieriga-4

Abstract

XieRiGa-4 (XRG-4) is a widely used formulation in Mongolian medicine for the treatment of kidney dysfunction, yet its active constituents and underlying mechanisms of action remain poorly characterized. This study aims to identify active constituents, key targets, and potential pharmacological mechanisms of XRG-4 in protecting against kidney injury using network pharmacology, molecular docking, and in vivo and in vitro validation. A rat kidney injury model was established by pylorus ligation to evaluate XRG-4 efficacy. Active components of XRG-4 were characterized by liquid chromatography-mass spectrometry (LCMS), and candidate targets were obtained from public databases. Network pharmacology analyses, including protein-protein interaction (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, were conducted to identify core bioactive components, potential targets, and signaling pathways. Molecular docking assessed binding affinities between active compounds and core targets, and Western blot analysis validated expression of key targets. Biochemical analysis revealed that serum creatinine (CRE) and urea (URE) levels were significantly higher in the model group than in the normal group (P < 0.01) and were significantly reduced after XRG-4 treatment (P < 0.01), indicating a renal protective effect. Cell viability was markedly decreased in the model group (P < 0.01) and significantly improved in XRG-4-treated groups (P < 0.01). A total of 20 bioactive compounds, including the top 10 cationic and anionic constituents, and 253 overlapping targets associated with XRG-4 and kidney injury were identified. Network analysis indicated a central role of the EGFR signaling pathway in XRG-4 efficacy. Molecular docking revealed that five key compounds, 6- Hydroxypurine, Hymecromone, Gardenoside, Geniposide, and Phenprobamate, displayed strong binding affinities with three core targets, Slc22a5, Mrps9, and EGFR. Western blot analysis further confirmed that Slc22a5 (P < 0.05) and Mrps9 (P < 0.01) expression was significantly reduced in the model group and markedly upregulated following XRG-4 treatment. This study confirms that Mongolian medicine XRG-4 improves renal function and alleviates kidney injury. Using network pharmacology, molecular docking, and experimental validation, its main active components were found to bind to core targets including EGFR, Slc22a5, and Mrps9, regulate the EGFR signaling pathway, and upregulate Slc22a5 and Mrps9 expression, thereby exerting a nephroprotective effect and providing a scientific basis for its clinical application. This study clarifies the nephroprotective effects of XRG-4 and demonstrates its capacity to attenuate kidney injury through the EGFR signaling pathway. These findings provide a scientific rationale for clinical use of XRG-4 in kidney diseases and support further development of XRG-4 as a potential nephroprotective agent in Mongolian medicine.