Nedd4 family interacting protein 1 (Ndfip1), an adaptor protein for Nedd4-mediated ubiquitination, has been found to play an essential role in regulating divalent metal transporter 1 (DMT1) and iron homeostasis in human cortical neurons.The present study is to investigate the effect of Ndfip1 on 6-hydroxydopamine (6-OHDA)-induced iron accumulation in MES23.5 dopaminergic cells and the possible mechanisms.Results showed the expressions of Ndfip1 decreased in 6-OHDA-treated rats and MES23.5 cells.DMT1 (+IRE) protein level and ferrous iron influx were increased in 6-OHDA treated MES23.5 cells.This induced a decrease in ΔΨm and an increase in ROS production after iron treatment.Over-expressed Ndfip1 reversed the above processes.These results suggest over-expression of Ndfip1 could decrease DMT1 (+IRE) protein level in 6-OHDA treated MES23.5cells and reduce DMT1 (+IRE)-mediated iron influx.This further suppressed iron-induced ROS generation and decrease of ΔΨm, and then protect the cells.