Ginkgolide B Promotes Angiogenesis After Oxygen-Glucose Deprivation by Regulating AKT1 in bEnd.3 Cells

Ischemic stroke leads to significant neuronal damage, and impaired angiogenesis remains a critical factor limiting post-stroke recovery. Ginkgolide B (GB), a key component of Ginkgo biloba extract, has shown potential neuroprotective effects, but its pro-angiogenic mechanisms remain unclear.

To investigate the effects of GB, we established an oxygen-glucose deprivation/reperfusion (OGD/R) model using bEnd.3 cells. Potential molecular targets of GB were explored through a combination of network pharmacology analysis, protein-protein interaction (PPI) network construction, pathway enrichment, and molecular dynamics simulations. Based on these predictions, a series of in vitro assays-including Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU) incorporation, wound-healing, Transwell migration, and Matrigel tube formation tests-were performed to evaluate cell viability, proliferation, migration, and angiogenic activity. Western blotting was conducted to detect AKT serine/threonine kinase 1 (AKT1), vascular endothelial growth factor (VEGF), and Angiogenin (Ang) expression and clarify the role of the AKT1/VEGF/Ang pathway.

Bioinformatics analysis identified 19 potential targets, among which AKT1, Matrix Metalloproteinase 9 (MMP9), and Prostaglandin-Endoperoxide Synthase 2 (PTGS2) exhibited the highest relevance. GB showed no evident cytotoxicity at concentrations up to 40 μM and mitigated the OGD/R-induced reduction in cell viability. At this concentration range, GB also enhanced endothelial proliferation, migration, and tube formation in bEnd.3 cells. Mechanistic studies revealed that MK2206 inhibition of AKT1 markedly suppressed AKT1 expression (p < 0.01), impaired angiogenic capacity, and aggravated ischemic-hypoxic injury, whereas GB treatment significantly increased VEGF and Ang expression (p < 0.01), likely via AKT1 upregulation (p < 0.01).

GB promotes angiogenesis and exerts neuroprotective effects by activating the AKT1/VEGF/Ang signaling pathway, suggesting its potential therapeutic value for ischemic stroke-related injuries.