Abstract: Objective: The purpose of this study was to examine how lotus seed extract (LSE) affected mice's hippocampus neuroprotection and anxiety-like behaviors. Methods: Uncertain empty bottle stress and chronic confinement stress were combined to create a mouse model of generalized anxiety disorder (GAD). The mice were split into the following groups: low-dose (9 mg·kg⁻¹·d⁻¹), medium-dose (18 mg·kg⁻¹·d⁻¹), and high-dose (36 mg·kg⁻¹·d⁻¹) LSE groups; control group; model group; and diazepam (DZP, 2 mg·kg⁻¹·d⁻¹) group. For 14 days in a row, treatments were given orally. The elevated plus maze (EPM) and open field test (OFT) were used to measure anxiety-like behaviors. ELISA was used to assess the amounts of 5-hydroxytryptamine (5-HT), norepinephrine (NE), and γ-aminobutyric acid (GABA) in plasma and hippocampus tissues. Quantitative real-time PCR and Western blot analysis were used to determine the mRNA and protein expression levels of brain-derived neurotrophic factor (BDNF), amyloid β-protein (Aβ), and tau in the hippocampus. Results: Mice in all LSE-treated groups showed improved coat condition, increased body weight, and increased food intake as compared to the model group. LSE can improve OFT and EPM behavioral indicators to varying degrees, with more significant improvements in the medium and high dose groups (P<0.01). In both plasma and hippocampus tissues, LSE intervention dose-dependently raised 5-HT and GABA levels while lowering NE levels (P<0.01). At the same time, LSE administration markedly increased the mRNA and protein expression of BDNF in the hippocampus (P<0.01) and decreased the mRNA and protein expression of Aβ and Tau (P<0.01). Conclusion: Lotus seed extract has anxiolytic and hippocampus neuroprotective effects. These effects may be achieved by blocking aberrant Tau aggregation and Aβ deposition, triggering the BDNF neurotrophic pathway, and adjusting the balance of 5-HT, NE, and GABA neurotransmitters.