Abstract: OBJECTIVE To investigate the inhibitory effect of aucubin on glutamate-induced excitatory neurotoxicity. METHODS A neurotoxicity model was established using 20 mmol·L^-1glutamate to induce damage in PC-12 cells. After treatment with 1, 5, 10, 20 and 40 μmol·L^-1 aucubin for 24 h, cell viability was measured by MTT, intracellular Ca²⁺ and reactive oxygen species(ROS) levels were measured by flow cytometry, the level of glutamate receptor N-methyl-D-aspartic acid receptor(NMDAR1) were measured by In Cell Western. And superoxide dismutase(SOD), malondialdehyde(MDA), and lactate dehydrogenase(LDH) levels in cell supernatants were measured by using ELISA kits. RESULTS After treated with 20 mmol·L^-1glutamate for 24 h, the viability of PC-12 cells was significantly reduced(P<0.01), the levels of intracellular Ca²⁺, NMDAR1 ROS, MOA, LDH were significantly increased (P<0.01), the level of SOD was significantly reduced. 10 μmol·L^-1 aucubin could significantly increase the cell viability of PC-12 cells injured by glutamate(P<0.01), it could reduce the levels of intracellular Ca²⁺, NMDAR1, ROS and LDH(P<0.01), and elevate intracellular SOD level(P<0.01). CONCLUSION Aucubin may ameliorate glutamate-induced damage in PC-12 cells by inhibiting NMDAR1 expression, thereby reducing the accumulation of intracellular Ca²⁺, and inhibiting oxidative stress levels to suppress glutamate-induced excitatory neurotoxicity.