Network pharmacology-based prediction and exploration of the mechanism of Bushen Mingmu Wan in glaucoma treatment

Introduction Glaucoma is an irreversible, sight-threatening optic neuropathy. Current therapies are limited by inadequate neuroprotection and ocular surface adverse effects. Traditional Chinese Medicine (TCM) presents unique advantages in glaucoma management. Bushen Mingmu Wan (BSMMW), a TCM formula that tonifies the liver and kidney, demonstrates clinical efficacy in preserving visual function in glaucoma patients; however, its underlying molecular mechanisms remain poorly defined. This study explored BSMMW's therapeutic mechanisms against glaucoma via network pharmacology and experimental validation. Methods Active compounds and targets of BSMMW were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and PharmMapper. Glaucoma-related targets were from GeneCards, the Online Mendelian Inheritance in Man (OMIM) database, the Therapeutic Target Database (TTD), and DrugBank. A protein-protein interaction (PPI) network was constructed using STRING and Cytoscape. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed via Metascape. Furthermore, BSMMW's therapeutic efficacy against glaucoma was validated in Sprague-Dawley (SD) rats and R28 cells using enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction, western blot, immunohistochemistry, and haematoxylin and eosin staining. Results A total of 118 overlapping targets shared between BSMMW compounds and glaucoma were identified by network pharmacology analysis. Core hub targets of BSMMW in glaucoma treatment, including ALB, interleukin-6 (IL-6), tumor necrosis factor (TNF), AKT serine/threonine kinase 1 (AKT1), and interleukin-1β (IL-1β), were revealed by PPI network and KEGG enrichment analyses; AKT1 and prostaglandin-endoperoxide synthase 2 (PTGS2) were further validated as key functional nodes in the integrated TCM component-target-pathway network. Significantly enriched pathways, including the phosphatidylinositol 3-kinase/Akt (PI3K/Akt), advanced glycation end products-receptor for advanced glycation end products (AGE/RAGE), mitogen-activated protein kinase (MAPK), and hypoxia-inducible factor-1 (HIF-1) signaling pathways, were observed. In vivo , retinal histopathological damage was alleviated and the upregulation of key inflammatory factors (IL-1β, IL-6, TNF-α) in retinal tissue and serum was suppressed by BSMMW, particularly at the high dose; consistent suppressive effects on these factors were observed in R28 cells in vitro . Conclusion BSMMW exerts therapeutic effects against glaucoma through a multi-component, multi-target, and multi-pathway mechanism, primarily by modulating anti-inflammatory responses, thus offering experimental evidence supporting its clinical use.