Introduction: Major depressive disorder is a prevalent psychiatric condition affecting millions worldwide, yet current pharmacotherapies, primarily based on monoamine modulation, are limited by delayed onset, insufficient efficacy, and adverse effects. Maizhuni-Nujia (MZNNJ) is a classic multi-herb prescription from Uyghur traditional medicine, traditionally used for melancholia and amnesia, but its antidepressant mechanisms and scientific basis remain largely unexplored. This study aimed to systematically investigate the antidepressant effects and underlying mechanisms of MZNNJ using a reserpine-induced acute depression model in mice.Methods: An acute depression model was established in mice by intraperitoneal injection of reserpine (4 mg/kg). Mice were pre-treated orally with various doses of MZNNJ (6.15, 12.3, or 24.6 g/kg), its individual herbal components (processed or unprocessed), or fluoxetine (4.1 mg/kg) for 14 days. Behavioral assessments included eyelid ptosis, the circle escape test, and the Morris water maze. Neurochemical markers (5-HT, DA, BDNF) were measured by ELISA, and histopathological changes in the hippocampus and cortex were evaluated with H&E staining. Proteomic and metabolomic profiling of brain tissue were performed using DIA-based mass spectrometry and UHPLC-MS/MS, respectively. Key proteins in the Ras/ERK/CREB/BDNF signaling pathway were validated by Western blotting and immunofluorescence.Results: MZNNJ treatment dose-dependently ameliorated reserpine-induced depressive-like behaviors, including ptosis, reduced escape behavior, and spatial learning and memory deficits. This was accompanied by significant restoration of 5-HT, DA, and BDNF levels in both serum and brain tissue, and marked improvement in neuronal damage in hippocampal subregions. Comparative analysis revealed that the formulation containing processed herbs (ACFI and MPF) was significantly more efficacious than the formulation containing raw herbs. Proteomics and subsequent validation showed that MZNNJ robustly reversed the reserpine-induced downregulation of KSR1, Ras, p-ERK, p-CREB, and BDNF. Metabolomics further revealed that MZNNJ corrected brain metabolic disturbances, with significant enrichment in pathways related to glutathione metabolism, energy metabolism, and the "Spinocerebellar ataxia" pathway.Conclusion: This study provides the first comprehensive evidence that MZNNJ exerts potent antidepressant effects through multifaceted mechanisms involving activation of the KSR1-mediated Ras/ERK/CREB/BDNF neurotrophic pathway and systemic restoration of brain metabolic homeostasis. These findings validate the traditional processing and compatibility principles of Uyghur medicine and position MZNNJ as a promising candidate for further development as a novel antidepressant therapy.