Based on network pharmacology and molecular docking, combined with in vitro experiments, to explore the mechanism of ursolic acid (UA) in treating glioblastoma (GBM). UA targets were obtained from TCMSP, SwissTargetPrediction, SuperPred databases, and disease targets of GBM were obtained from four disease databases including GeneCards, DisGeNET, TTD, and PharmGkb combined with two GEO gene chips. Cytoscape software was used to screen the core target of UA in the treatment of GBM by protein interaction. GEPIA (gene expression profiling interactive analysis) database was used to analyze whether there is a significant difference in mRNA expression levels of core targets between GBM patient tissues and normal tissues. David database was used to carry out GO (gene ontology) function and KEGG (Kyoto encyclopedia of genes and genomes) pathway enrichment analysis for potential targets. Autodock Vina and Pymol were used to realize the molecular docking of UA and the significant core proteins. Finally, the anti-tumor activity of UA was further verified through in vitro experiments. The results indicated that 92 protein targets corresponding to UA were screened, and 7 433 targets of GBM were obtained, and 72 potential targets were obtained after combination. The PPI network and topology analysis results show that 10 core targets, including CCND1, EGFR, FOS, CASP3, HSP90AA1, IL6, CTNNB1, INS, RELA, and VEGFA are the core targets. The expression of CASP3, CCND1, EGFR, FOS, IL-6, and VEGFR mRNA in GBM tissue was significantly higher than that in normal tissue by GEPIA database analysis. GO enrichment analysis found that UA treatment of GBM was closely related to cellular response to chemical stress, membrane raft, cysteine-type endopeptidase activity, etc. KEGG pathway analysis showed that it involved apoptosis, TNF signaling pathway, PI3K-Akt signaling pathway, etc. The results of in vitro experiments showed that UA could inhibit the proliferation of U251 cells, The IC50 value is 26.66 μmol/L. The results of RT-qPCR and WB(western blot) showed that UA could regulate the expression of CASP3 and CCND1 mRNA and protein in U251 cells, and UA could promote the overexpression of CASP3 mRNA and protein and inhibit the expression of CCND1 mRNA and protein in U251 cells. UA can treat glioblastoma through multiple targets and pathways.