Glioblastoma (GBM) is the most prevalent and aggressive primary brain tumor and associated with a poor prognosis. Guizhi Fuling Wan (GZFLW), a modernized traditional Chinese medicine preparation, has shown clinical promise in improving outcomes for patients with GBM, however, its active ingredients and underlying mechanisms remain unclear. This study aimed to identify the active ingredients of GZFLW and investigate its potential mechanisms in GBM treatment using network pharmacology analysis combined with in vitro validation . Network pharmacology analysis, molecular docking, and experimental validation were employed to identify the principal active compounds in GZFLW and to predict their primary targets in GBM. Cellular experiments using U87-MG and U251-MG cells were performed to further validate their therapeutic potential and mechanism in GBM. Network pharmacology analysis and experimental verification suggested that quercetin, kaempferol, baicalein, and ellagic acid were the top four key active components. Additionally, HIF1A, MAPK1, and AKT emerged as the key targets of GZFLW against GBM. In vitro experiments demonstrated that GZFLW suppressed GBM cell proliferation by reducing colony-forming ability. Mechanistically, it induced G0/G1 phase arrest through p21-mediated regulation of the CDK2–cyclin E/cyclin A2 and CDK4/ 6–cyclin D1 complexes, and promoted apoptosis, likely via the mitochondria-mediated apoptotic pathway. Network pharmacology integrated with cellular experimental further validated the predicted anti-GBM mechanisms of quercetin, kaempferol, baicalein, and ellagic acid at the molecular and cellular levels. These findings provide a scientific foundation and valuable reference for future research on the role of GZFLW in GBM treatment.
Zhu et al. (Wed,) studied this question.