Glioblastoma (GBM) is a malignant tumor with a high incidence rate and high invasion in human central nervous system tumors. However, the causes of the occurrence and development of glioblastoma are not clear. Although Ginkgo Folium (GF) extract can reduce the growth of U-87 glioma cell lines in nude mice, the action mechanism remains unclear. In this study, network pharmacology, molecular docking, and cell experiment were utilized to explore the underlying molecular mechanism of GF in treating GBM. The effective ingredients and action targets of GF were obtained from the TCMSP database. STRING database was applied for protein interaction analysis and Cytoscape software was used to build the "compound-target-disease" network and PPI network. The R language was adopted as gene ontology(GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. AutoDock software was used for molecular docking verification. A cell experiment was employed to verify the inhibition of GBM by the effective compounds in GF. The results showed that 27 potential effective components and 112 potential targets of GF were selected. 7 435 targets of GBM were screened from six databases, and 98 overlapping targets were obtained. In active component-target network, quercetin, kaempferol, stigmasterol, and luteolin were the important active components. In PPI network, ESR1, RELA, FOS,CCD1, EGFR, AR, HIF1A, MAPK8, NCOA1, and MDM2 were the core targets. GO and KEGG analyses showed that the main pathways of GF in treating GBM involved PI3K-Akt, MAPK, and TNF signaling pathway. The results of molecular docking verified the good binding activity of the key pharmacodynamic molecules with the core targets. Cell experiments showed that quercetin (QUE), luteolin (LUT), and kaempferol (KAE) can inhibit cell viability in GBM cell lines U251. It can be concluded GF is the result of multi-component, multi-target, and multi-pathway interaction in the treatment of GBM, which provided a theoretical basis for the clinical application of GF in the treatment of GBM.