It has been suggested that oxidative stress of reactive oxygen species(ROS) may play an important role in the pathogenesis of neurological disorder. The aim of this study was to elucidate the oxidative stress of glucose oxidase(GO) in the cultured mouse spinal motor neurons and the preventing effect of Benincasae Semen(BS) on ROS-induced neurotoxicity. Cytotoxic effect of GO and protective effect of BS were performed by MTT assay. 30mU/ml GO decreased cell viability in dose-and time-dependent mannner, and BS diminished GO-induced neurotoxicity in these cultures. From above the results, ROS such as GO has toxic effect, and herb extract of BS is very effective against GO-induced neurotoxicity in cultured spinal motor neurons of neonatal mouse.