Background: Transient receptor potential vanilloid 4 (TRPV4) is a nonselective calcium-permeable cation channel. Our previous study revealed that TRPV4 mediates endoplasmic reticulum (ER) stress and inflammation, leading to the loss of dopamine neurons in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease (PD) model mice. Objective: The aim of the present study was to investigate the molecular role of TRPV4 in ferroptosis in PD models. Methods: We used GSK2193874 (an inhibitor of TRPV4) and siRNA targeting TRPV4 to explore the role of TRPV4 in iron transportation, the production of reactive oxygen species (ROS) and oxidative stress in 1-methyl-4-phenylpyridinium ion (MPP+)-treated PC12 cells by H2DCFDA, CCK-8, ELISA and western blot. In a PD mouse model, the intracerebral injection of adeno-associated virus (AAV) was used to knockdown or upregulate TRPV4 expression following the intraperitoneal injection of MPTP and GSK2193874. We used Perl's iron staining and western blot to detect alterations in iron-positive cells and ferroptosis-associated molecules in the substantia nigra (SN). Results: We found that silencing TRPV4 increased the expression of glutathione peroxidase 4 (GPX4) and ferritin heavy chain (FTH1), decreased divalent metal transporter 1 (DMT1), and alleviated the excessive ROS and oxidative stress in MPP+-induced PC12 cells. Moreover, TRPV4 regulated the accumulation of iron in the SN and ferroptosis-associated signalling pathways in PD mice. Conclusions: TRPV4 plays a key role in MPP+/MPTP-induced ferroptosis. Our results indicate that the downregulation of TRPV4 may a represent promising treatment for PD through the inhibition of ferroptosis.