Study on the Active Pharmaceutical Substances and Potential Quality Markers of Yueju Wan Antidepressant Based on Metabolomics

AIM OF THE STUDY: Integrating metabolomics, network pharmacology, and zebrafish depression models, this study explored the pharmacodynamic material basis and potential quality markers (Q-markers) of Yueju Wan (YJW) against depression. METHODS: Chronic unpredictable mild stress (CUMS) was adopted to establish a rat depression model for evaluating the antidepressant efficacy of YJW. UPLC-QTOF-MS/MS was utilized to characterize the chemical constituents of YJW, and relevant databases were employed to predict its potential therapeutic targets. Metabolomics analysis was performed to screen differential metabolites in serum and urine, followed by pathway enrichment analysis. A compound-target-metabolite-pathway interaction network was constructed to screen core therapeutic targets and candidate pharmacodynamic substances, and RT-qPCR, Western blot, and zebrafish depression models were applied to validate the identified targets and pharmacological effects. Finally, a quantitative detection method for the candidate quality markers was established. RESULTS: One hundred eight chemical components and 105 core targets were characterized and predicted from YJW, respectively. YJW reversed 87 depression-associated differential metabolites. Further integrated analysis together with RT-qPCR and Western blot validation suggested that Alox15 and Cyp19a1 may be involved in the antidepressant-like effects of YJW, possibly through modulation of arachidonic acid metabolism, microglial polarization, and neuroinflammatory responses. Seventeen compounds including α-cyperone, geniposide, ferulic acid, and atractylodin were verified to exert antidepressant activities via molecular docking and zebrafish models. Ferulic acid, α-cyperone, geniposide, and atractylodin show satisfactory detectability and are proposed as potential Q-markers. CONCLUSIONS: This study provides new evidence for the candidate antidepressant pharmacodynamic substances and potential quality markers of YJW and provides a scientific basis for the new drug development and quality control of YJW.