Abstract
BACKGROUND: Angiogenesis is a fundamental physiological process mediating vascular network formation, represents a critical therapeutic target for ischemic diseases and tumor neovascularization. Xuefu Zhuyu decoction (XFZYD), a classical formula for promoting blood circulation and resolving stasis, demonstrates pro-angiogenic effect with safflower functioning as the sovereign herb. Hydroxysafflor yellow A (HSYA), the primary bioactive constituent of safflower, exerts potent angiogenesis modulation, defining its pharmacological significance. METHODS: In this study, in vitro tubulogenesis assay and cytocompatibility analysis were employ on human microvascular endothelial cell (HMEC-1), followed by target prediction via network pharmacology and molecular docking; immunoblotting analysis was performed to experimentally validate the pro-angiogenic molecular mechanism of HSYA. RESULTS: HSYA exerted concentration-dependent pro-angiogenic effects on HMEC-1 cells over 24 h without compromising cell viability (p > 0.05) across 0-200 μM. 121 potential targets of HSYA within the angiogenesis regulatory network were identified. Functional enrichment analysis revealed fluid shear stress, lipid metabolism, HIF-1, PI3K-Akt, and VEGF signal pathways as primary regulatory pathways. 8 hub targets derived from the protein-protein interaction (PPI) network were subjected to molecular docking. High-affinity interactions were observed for key angiogenesis regulators: MMP9 (-7.6 kcal·mol(-1)), and HIF-1α(-4.5 kcal·mol(-1)), which were functionally validated by immunoblotting analysis, preliminary demonstrating the mechanism of HSYA-mediated angiogenesis promotion. CONCLUSION: HSYA demonstrates significant pro-angiogenic activity on HMEC-1. Mechanistically, HSYA modulates multiple signaling pathways, with HIF-lα and MMP9 demonstrating regulatory significance. These findings suggest a molecular basis for HSYA's therapeutic potential in ischemic vascular pathologies.