Objective We explored the anti-depressant activity and mechanism of arecoline in vivo in a mouse model of depression induced by chronic unpredictable mild stress. The aim was to explore the possible mechanisms of action, providing experimental evidence for further research into the health benefits of arecoline and theoretical support for the scientific development and utilization of this resource. Methods Sixty quarantine-qualified SPF C57BL/6J mice were divided randomly into a control group, model group, fluoxetine group (20 mg/kg), and arecoline low-, medium-, and high-dose groups (10, 20, and 40 mg/kg, respectively) according to body mass (n=10 mice per group). The effects of arecoline on the behavior of the mice were evaluated by open-field, tail suspension, and forced-swimming tests. Serum corticosterone and serum and brain levels of superoxide dismutase (SOD) were detected by enzyme-linked immunoassay. Malondialdehyde (MDA), catalase (CAT), 5hydroxytryptamine (5-HT), and norepinephrine (NE) levels in brain tissue, and dopamine (DA), gamma aminobutyric acid (GABA), tumor necrosis factor (TNF-α), interleukin (IL)-10, IL-1β, brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), and cAMP-response element binding protein (CREB) were detected by Western Blot. Results Arecoline significantly reduced the total distance and average speed of the model mice in open field tests and increased activities, and significantly reduced the immobility time in the tail suspension and forced swimming tests. Arecoline also significantly decreased serum corticosterone levels, increased SOD and CAT, and decreased MDA levels. 5-HT, DA, NE, and GABA levels were significantly increased, and the cytokines TNF-α, IL-6, and IL-1β were significantly decreased. Expression levels of BDNF, TrkB, and CREB in the brain tissue were significantly increased. Conclusions Research has found that arecoline has a significant antidepressant ability, and its mechanism may be achieved by reducing oxidative stress damage, inhibiting neuroinflammation, regulating neurotransmitter balance, and regulating the BDNF/TrkB/CREB signaling pathway. . This study explored the antidepressant efficacy of arecoline and preliminarily revealed its possible regulatory mechanism, which can provide data support for the neuroactivity of arecoline and lay a theoretical foundation for the development of arecoline as medicine.