目的 评价柴胡疏肝散（CSP）对抑郁症的药效学影响，并探究CSP治疗抑郁症的关键有效入脑成分及潜在作用机制。方法 制备慢性社交挫败应激（CSDS）小鼠模型，模型组、CSP（6.46 g·kg^-1）组和氟西汀（10 mg·kg^-1）组分别ig给药12 d，通过行为学测试评估各组小鼠的社交能力以及抑郁程度；苏木素-伊红（HE）、尼氏染色观察海马神经元与尼氏体状态；采用超高效液相色谱-四极杆-飞行时间串联质谱（UPLC-Q-TOF-MS）技术全面分析CSP的原型入脑成分。通过Pubchem、SwissTargetPrediction查找CSP原型入脑成分对应的靶点，通过GeneCards查找抑郁症相关基因，绘制疾病与入脑成分相关基因的Venn图。根据成分及作用靶点制作成分-靶点网络图。将STRING数据库导出的靶点蛋白质-蛋白质相互作用（PPI）数据导入Cytoscape3.9.1软件，构建PPI网络，调用Centiscape 2.2插件对网络节点进行拓扑分析，计算各节点的关键中心性参数，以各参数的平均值为筛选阈值得到同时满足3项阈值条件的节点，作为核心靶点。使用微生信对核心靶点进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集分析。最后建立神经元应激模型，以CCK-8法、TUNEL染色法、JC-1试剂盒和Western blotting法对关键有效入脑成分柴胡皂苷A的作用机制进行验证。结果 CSP对小鼠的抑郁样行为和海马神经元具有改善保护作用。利用网络药理学对UPLC-Q-TOF-MS鉴定出的9个原型入脑成分治疗抑郁靶点进行预测，聚焦到柴胡皂苷A是CSP中发挥抗抑郁作用的关键入脑成分，其作用机制可能与改善细胞凋亡有关，最后通过体外实验对其进行验证，从机体-细胞-分子层面，得出柴胡皂苷A通过对线粒体膜电位产生影响，并上调B细胞淋巴瘤-2（Bcl-2）的表达，下调Bcl-2相关X（Bax）、半胱天冬氨酸蛋白酶-3（Caspase-3）和Caspase-9的表达发挥抗抑郁的作用。结论 CSP中发挥抗抑郁的关键入脑成分为柴胡皂苷A，其机制与改善细胞凋亡有关。

Objective To evaluate the pharmacodynamic effects of Chaihu Shugan Powder (CSP) in treating depression and to identify its key brain-penetrating active components, as well as to elucidate the underlying mechanisms. Methods A chronic social defeat stress (CSDS) mouse model was established, and the model group, CSP group (6.46 g·kg^-1), and fluoxetine group (10 mg·kg^-1) were administered ig for 12 d, respectively. Depression-like behaviors and social ability were assessed using a battery of behavioral tests. Hippocampal neuronal morphology and Nissl body status were examined by hematoxylin-eosin (HE) and Nissl staining. The prototype brain-penetrating components of CSP were comprehensively analyzed using ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS). Putative targets of these components were retrieved from PubChem and SwissTargetPrediction databases, while depression-related genes were obtained from GeneCards. A Venn diagram was generated to identify overlapping genes between CSP components and depression. A component-target network was constructed, and protein-protein interaction (PPI) data retrieved from the STRING database were imported into Cytoscape 3.9.1 to establish a PPI network. Topological analysis was performed using the Centiscape 2.2 plugin, and nodes satisfying all three predefined threshold criteria were defined as core targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted using the microbioinformatics platform. Finally, a neuronal stress model was established, and the mechanism of the key brain-penetrating component, saikosaponin A, was validated using CCK-8 assay, TUNEL staining, JC-1 kit, and Western blotting. Results CSP significantly ameliorated depression-like behaviors and protected hippocampal neurons in mice. Network pharmacology analysis of the nine prototype brain-penetrating components identified by UPLC-Q-TOF-MS revealed that saikosaponin A was the key antidepressant component, and its mechanism was closely associated with the regulation of apoptosis. Subsequent in vitro validation confirmed that saikosaponin A exerted its antidepressant effects by modulating mitochondrial membrane potential, upregulating Bcl-2 expression, and downregulating the expression of Bax, Caspase-3, and Caspase-9 at both the organismal, cellular, and molecular levels. Conclusion The key brain-penetrating component responsible for the antidepressant action of CSP is saikosaponin A, and its therapeutic mechanism involves the inhibition of neuronal apoptosis.

R285.5