Objective To elucidate the molecular mechanism of reserpine-induced depression using network toxicology, molecular docking techniques, behavioral assessments of animal models, and histopathological analyses. Methods Core targets were screened using multi-database network toxicology, followed by the construction of a protein-protein interaction network and validation of core targets through molecular docking. Sprague Dawley rats were divided randomly into control and reserpine (0. 5 mg / kg) groups, and administered the corresponding treatments once daily for 4 consecutive days. Behavioral changes were assessed using the forced-swim and open-field tests. Serum neurotransmitters were quantified by enzyme-linked immunosorbent assay and neuropathological damage was observed via tissue staining. Target gene expression regulation was verified by Western blot. Results Network toxicology screening and molecular docking simulation demonstrated that reserpine exhibited significant binding affinity with the dopamine D2 receptor, cyclic-AMP response element binding protein, and serine / threonine-protein kinase 1 (AKT1). Animal experiments demonstrated that reserpine-treated rats displayed depression-like behaviors, including motor inhibition (P<0. 01), with decreased serum levels of norepinephrine and 5-hydroxytryptamine ( P<0. 01), respectively. Pathological observations revealed microglial proliferation in the cerebral cortex, increased apoptosis, and reduced Nissl bodies in the hippocampal CA1 region. Down-regulation of brain-derived neurotrophic factor (BDNF) in brain tissue and decreased expression of hippocampal AKT1 and phosphorylated AKT1 were also observed. Conclusions Reserpine influences monoamine transmitter metabolism and neuronal structural integrity via the inhibition of BDNF and AKT1 protein expression, resulting in depressive-like behavior and cerebral nerve damage in rats.