Introduction and objective:
Ferroptosis is essential for the occurrence and development of myocardial infarction. The aim of the study is to elucidate the molecular mechanism of Ginkgolide Injection against ischemic cerebral infarction (ICI), focusing on its multi-target regulation of ferroptosis via the AKT1 and NFE2L2.

Material and methods:
Human primary neurons and astrocytes were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to mimic cerebral ischemia. Cells were treated with Ginkgolide Injection or transfected with AKT1-targeting siRNA. Targets of Ginkgolide Injection components against ICI-associated genes were retrieved from TCMSP, CTD, HERB, OMIM, and DisGeNET databases. A compound-target-disease network was constructed using Cytoscape, followed by PPI, GO, and KEGG analyses. Cell viability (CCK-8), lipid peroxidation assessment (C11-BODIPY581/591 staining), redox markers (GSH, ROS, MDA), and AKT1/Nrf2 signaling (RT-qPCR, Western blot) were analyzed.

Results:
Ginkgolide Injection restored viability in OGD/R-injured neurons and astrocytes. Bilobalide in Ginkgolide Injection targeted 22 genes; Ginkgolide A targeted 32 genes; Ginkgolide B targeted 48 genes; Ginkgolide C targeted 30 genes; Ginkgolide J targeted 7 genes. A total of 55 targets for Ginkgolide Injection were common with ICI-associated genes. Network pharmacology identified AKT1 and NFE2L2 as core targets of Ginkgolide Injection against ICI. Ginkgolide Injection suppressed ferroptosis (reduced PTGS2 mRNA and lipid peroxidation) and activated AKT1 signalling in OGD/R-injured neurons and astrocytes. AKT1 knockdown abolished Ginkgolide’s anti-ferroptotic effects, confirming AKT1’s regulatory role.

Conclusions:
Ginkgolide Injection mitigates cerebral infarction by co-activating AKT1 and NFE2L2 to suppress ferroptosis, highlighting its potential as a multi-target therapy for ischemic ICI.