OBJECTIVE To investigate the effect and potential mechanism of Ziyin Mingmu Formula in treating age-related macular degeneration (AMD) by combining network pharmacology with animal model validation.

METHODS Active ingredients of Ziyin Mingmu Formula were obtained from the TCMSP and BATMAN databases, and their targets were searched using the PharmMapper database. AMD disease targets were identified using the GeneCards, DrugBank, OMIM, and TTD databases. Venny analysis was performed to identify the intersection of active ingredient and disease targets. A protein-protein interaction (PPI) network was constructed using the String database, and core targets were screened using Cytoscape 3.9.0 software. KEGG pathway enrichment analysis was performed using the DAVID database. Molecular docking of key active ingredients with core targets was performed using Autodock software. A laser-induced mouse choroidal neovascularization (CNV) model was used. Optical coherence tomography angiography (OCTA) was used to assess CNV area in vivo, immunofluorescence staining was used to assess CNV area on choroidal flat mounts, and Western blot analysis was used to examine the expression of proteins involved in the vascular endothelial growth factor (VEGF) pathway.

RESULTS Network pharmacology analysis identified 221 active ingredients in the Ziyin Mingmu Formula. PPI analysis identified 29 core targets, including SRC, protein kinase B (AKT1), mitogen-activated protein kinase 1 (MAPK1), and heat shock protein 90α family class A member 1 (HSP90AA1). KEGG analysis revealed that the VEGF signaling pathway was the most highly enriched. Molecular docking revealed that the core targets SRC, AKT1, MAPK1, and HSP90AA1 had good binding affinity with the main active ingredients, diosmetin, catechin, vestitol, and licochalcone A. Animal experiments showed that the Ziyin Mingmu Formula significantly reduced CNV area in model mice, downregulated VEGF protein expression, decreased VEGFR2, p38, and ERK1/2 protein phosphorylation levels, and inhibited the VEGF signaling pathway.

CONCLUSION Ziyin Mingmu Formula may inhibit CNV formation by regulating the VEGF signaling pathway.