Objective To investigate the beneficial effects of catalpol-paeoniflorin ( CTP-PNF ) on perimenopausal depression (PMD) and to explore its underlying mechanism, specifically focusing on its ability to inhibit miR-124-to promote the proliferation and migration of neural stem cells. Methods A PMD mouse model was established, combined with intracerebral stereotaxic microinjection of miR-124 inhibitor and in vitro neural stem cell experiments. Depressive-like behaviors were evaluated using the sucrose preference test (SPT), tail suspension test (TST), and novelty-suppressed feeding test (NSFT). Relative expression levels of miR-124 in the prefrontal cortex of mice were detected by quantitative reverse transcription-polymerase chain reaction and levels of the neurotransmitters norepinephrine, dopamine, serotonin, and gamma-aminobutyric acid were measured by enzyme-linked immunosorbent assay. Neuronal pathological changes were observed by Nissl staining. The viability and migration ability of neural stem cells were detected by MTT and Transwell assays, respectively. The core targets and pathways involved in miR-124-mediated intervention of PMD by CTP-PNF were analyzed by network pharmacology. Results Compared with the model group, CTP-PNF significantly improved depressive-like behaviors in PMD mice and downregulated the expression of miR-124 in the prefrontal cortex (P<0. 001). Inhibition of miR-124 reduced depressive-like behaviors, increased neurotransmitter levels ( P< 0. 05 ), ameliorated the pathological status of the prefrontal cortex and hippocampus, and synergistically enhanced the viability and migration of neural stem cells with CTP-PNF-containing serum, thereby promoting neural stem cell repair. Comprehensive bioinformatics analysis based on network pharmacology revealed that miR-124-mediated treatment of PMD by CTP-PNF mainly involved 56 core therapeutic targets, including the hypoxia-inducible factor 1 signaling pathway, 5-HT signaling pathway, and cAMP signaling pathway related to mitogen-activated protein kinase 1, signal transducer and activator of transcription 3, TP53, and vascular endothelial growth factor A. Conclusions CTP-PNF significantly ameliorates depressive-like behaviors and neuropathological damage in PMD mice by downregulating miR-124 to promote the proliferation and migration of neural stem cells and regulate neurotransmitter networks. Its mechanism involves multiple signaling pathways, including hypoxia-inducible factor 1 and 5-HT. CTP-PNF may thus provide a new traditional Chinese medicine intervention strategy and molecular target for the treatment of PMD.