Fuzheng Yiai Formula and its Key Components Inhibited PCa by TP53/SLC7A11/GPX4-Induced Ferroptosis

Fuzheng Yiai Formula (FZYAF) is a traditional Chinese medicine compound, and its key components, ursolic acid (UA) and Anhydroicaritin (AHI), have potential antitumor activity, but their mechanisms of action in Prostate cancer (PCa) remain unclear. Key components and targets of FZYAF were identified by network pharmacology, and the components were further validated by HPLC-MS analysis. PCa cells were treated with UA and AHI to assess their effects on cell proliferation, migration, and ferroptosis. The interaction of UA or AHI with TP53 was assessed by molecular docking and DARTS. In vivo, a PCa mouse model was established, and tumor tissues from FZYAF-treated mice were subjected to metabolomic analysis. Additionally, the anti-tumor effects of FZYAF, UA, and AHI, and their impact on ferroptosis and the TP53 pathway were evaluated. Network pharmacology and HPLC-MS analysis, UA and AHI were identified as the key active components of FZYAF, and TP53 was predicted as a core target. In vitro, UA and AHI inhibited PCa cell proliferation and migration and induced ferroptosis, with their combination showing enhanced efficacy compared to either agent alone. Mechanistically, UA and AHI were bound to and upregulated the TP53 protein. In vivo, FZYAF inhibited tumor growth and induced ferroptosis in PCa mice, which was associated with alterations in lipid metabolism-related pathways, as revealed. UA and AHI, key components of FZYAF, inhibit PCa progression by binding to and upregulating the TP53 protein to induce ferroptosis. This study elucidates the role of FZYAF in developing anti-PCa drugs.