To investigate whether a standardized music-based intervention (MBI) improves post-stroke depression (PSD)–like behaviors and to elucidate hippocampal mechanisms focusing on neuroinflammation, neurotransmitter metabolism, and microglial reactivity.

PSD affects nearly half of stroke survivors and worsens motor and cognitive recovery. While MBIs are low-cost and safe, their mechanistic effects in PSD remain unclear. Understanding how music influences neuroimmune signaling may provide a novel non-pharmacologic approach for stroke rehabilitation.

Male C57BL/6J mice underwent middle cerebral artery occlusion followed by chronic unpredictable mild stress. Groups included Sham, Stroke, PSD, PSD + MBI (Mozart K.448, 60 min/day for 4 weeks), and PSD + Fluoxetine. Behavioral tests included modified neurological severity score (mNSS), sucrose preference (SPT), open-field (OFT), and Morris water maze (MWM). Hippocampal RNA-seq, LC-MS/MS proteomics, and neurotransmitter metabolomics were performed. Cytokines (TNF-α, IL-1β, IL-6, IL-10) were measured by qPCR/ELISA; microglia were examined by Iba1/CD86/CD206 staining, 3D morphometry.

Compared with PSD mice, MBI-treated animals showed lower mNSS scores ( p < 0.01), higher sucrose preference (p < 0.01), and improved spatial memory in MWM. Hippocampal 5-HT and NE were restored, and kynurenine/tryptophan ratios were decreased. MBI downregulated TNF-α, IL-1β, and IL-6 (p < 0.05) while increasing IL-10. Microglia displayed greater branch length and reduced soma volume, consistent with M2-like transformation. Multi-omics integration revealed suppression of NF-κB and MAPK signaling and activation of BDNF-TrkB pathways. Network analysis identified TREM2, CX3CR1, and NFKBIA as key regulatory hubs linking inflammation and neuroplasticity.

The results demonstrate that MBI mitigates PSD-like behaviors by attenuating hippocampal neuroinflammation, regulating neurotransmitter metabolism, and promoting microglial remodeling. These findings support the translational potential of music therapy as an adjunct to post-stroke neurorehabilitation.