Objective: To explore the effect and mechanism of baicalin on L-Glutamate-induced oxidative stress injury in the hippocampus of mice. Methods: Forty mice were randomly assigned to five groups: Sham, Model, N-Acetyl-L-Cysteine (NAC), and baicalin (BA-7.5mg/kg and BA-15mg/kg). A model of excitatory amino acid toxicity with oxidative stress injury was induced by injecting L-Glutamate into the lateral ventricle. The drugs were then injected intraperitoneally. Six hours later, behavioral tests were performed. The lesions of brain were observed using HE staining, while the apoptosis of neurons was assessed through TUNEL staining. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were determined using biochemical methods. The expression of Cytochrome C (CytC) was assessed by immunohistochemistry. Fluorescent staining was employed to detect the expression of reactive oxygen species (ROS). The levels of Nrf2, HO-1, SOD2 and Catalase (Cat) were detected by qPCR and WB. Results: The behavioral tests showed that the motion distance and pain threshold were reduced. MDA, ROS, and CytC were increased, SOD and Cat were decreased. The CA3 region of the hippocampus exhibited pathological changes, and the rate of TUNEL-positive increased. Baicalin could reverse these changes, especially BA-7.5mg/kg. Conclusion: Baicalin can reduce the hippocampal injury induced by L-Glutamate. This may be related to the activation of the Nrf2/HO-1 signaling pathway.