Modified Jie-Yu-He-Huan Capsule in Post-Stroke Depression Treatment: Insights from DIA Proteomic and PRM Validation

Background and Purpose: Post-stroke depression (PSD), a prevalent neuropsychiatric complication of stroke, manifests as persistent low mood and depressive symptoms secondary to cerebrovascular injury, contributing to increased morbidity and mortality. Although the modified Jie-Yu-He-Huan (MJYHH) capsule has demonstrated clinical efficacy in alleviating PSD symptoms, its pharmacological targets and mechanisms of action remain unclear. Methods: Pharmacological screening was performed on healthy C57BL/6 mice, revealing significant antidepressant effects of MJYHH. A PSD rat model was subsequently established through combined middle cerebral artery occlusion (MCAO), social isolation, and chronic mild stress (CMS). Behavioural tests were conducted to evaluate therapeutic outcomes. Data-independent acquisition (DIA) and parallel reaction monitoring (PRM) technologies were employed to investigate the traditional Chinese medicine (TCM) mechanisms. Results: The PSD model group exhibited characteristic depressive behaviours, which were significantly attenuated by MJYHH treatment. Proteomic analysis identified 16 differentially expressed proteins (DEPs), with subsequent Gene Ontology (GO), KEGG pathway, and protein-protein interaction (PPI) network analyses demonstrating their predominant association with: biosynthetic and metabolic pathways, oxidative stress response, mitochondrial dysfunction.PRM validation and molecular docking studies further implicated three key targets (Rrm2b, Cyp51a1, and Rhot2) as potential mediators of MJYHH's therapeutic effects. Conclusion: This integrated investigation combining animal models, omics technologies, and computational approaches elucidates the multi-target mechanism of MJYHH capsule in PSD treatment, providing a scientific foundation for TCM-based intervention strategies.