目的 探究柴胡疏肝散治疗抑郁症的药效物质及作用机制并进行实验验证。方法 应用超高效液相色谱-四极杆飞行时间质谱联用（UPLC-Q-TOF-MS/MS）技术对柴胡疏肝散化学成分进行分析；采用中药系统药理学数据库及分析平台（TCMSP）和SwisTargetPrediction数据库获取并筛选成分靶点；运用Limma包和hclust函数对GSE19738基因芯片数据进行差异基因分析和加权基因共表达网络（WGCNA）分析，获取抑郁症相关靶点，并与成分靶点取交集。绘制成分-靶点网络图，计算度值，筛选药效物质。交集基因通过基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）通路富集分析，随后进行最小绝对值收敛和选择算子（LASSO）回归分析和支持向量机（SVM）分析，得到柴胡疏肝散治疗抑郁症的潜在靶标，并进行ROC生存分析和分子对接验证以及基于斑马鱼抑郁模型的实验验证。结果 共从柴胡疏肝散水提物中鉴定126个成分，“成分-抑郁症”交集靶点共114个，度值≥11（中位数）的潜在药效物质44个，排名前5的成分为柴胡皂苷 A、川陈皮素、异甘草素、洋川芎内酯 A 和柚皮苷查尔酮。获得 5 个关键基因 FOS、TNF、NF-κB1、CXCR2 和IDO1，作为柴胡疏肝散抗抑郁症的潜在靶标。分子对接结果显示，5个关键成分与 5个潜在靶标均具有良好的结合能，其中与 NF-κB1和 TNF结合能最佳。通过体内实验研究，在确定柴胡疏肝散抗抑郁有效的前提下，柴胡疏肝散可显著降低斑马鱼抑郁模型体内促炎因子白细胞介素6（IL-6）和肿瘤坏死因子-α（TNF-α）的表达水平（P＜0.001）。在基因水平上，柴胡疏肝散可显著降低斑马鱼抑郁模型 NF-κB1、TNF-α和 IL-6 mRNA的表达水平（P＜0.05）。结论 柴胡疏肝散可通过柴胡皂苷A、川陈皮素等多种成分调控TNF-α/NFκB信号通路发挥抗抑郁作用。

Objective To explore the pharmacodynamic substances and mechanism of Chaihu Shugan Powder (CHSGP) in the treatment of depression and to verify the experimental results. Methods The chemical constituents of CHSGP were analyzed by UPLC-Q-TOF-MS/MS. Traditional Chinese Medicine System Pharmacology Database (TCMSP) and SwisTargetPrediction database were used to obtain and screen the component target. Differential gene analysis and weighted gene co-expression network analysis (WGCNA) analysis were performed on GSE19738 gene chip data using Limma package and hclust function, and depression related targets were obtained and intersected with component targets. The chemical composition-target network diagram was drawn, the degree value was calculated, and the pharmacodynamic substances were screened. The intersection genes were concentrated by GO and KEGG, followed by LASSO regression analysis and SVM analysis, to obtain the potential targets of CHSGP in the treatment of depression, and ROC survival analysis and molecular docking verification as well as experimental verification based on zebrafish depression model. Results A total of 126 chemical constituents were identified from CHSGP water extracts, 114 intersections of "chemical-depression" were identified, and 44 potential pharmacodynamic substances with a degree ≥ 11 (median) were identified. The top 5 components were saikosaponin A, nobiletin, 2',4,4'-trihydroxy-chalcone, ligustrolide A and naringin chalcone. Five key genes, FOS, TNF, NF- κB1, CXCR2 and IDO1, were obtained as potential targets for GHSGP antidepression. Molecular docking results showed that the five key components had good binding energy with five potential targets, and the binding energy with NF-κB1 and TNF was the best. In vivo studies, under the premise of determining the efficacy of CHSGP as an antidepressant, CHSGP can significantly reduce the expression levels of pro-inflammatory factors IL-6 and TNF-α in zebrafish depression models (P <0.001). At the gene level, CHSGP could significantly reduce the mRNA expression levels of NF-κB1, TNF-α and IL-6 in zebrafish depression model (P <0.05). Conclusion CHSGP can regulate the TNF-α/NF-κB signaling pathway through a variety of components such as saikosaponin A and chenorin to play an antidepressant role.

R285.5

黑龙江省重点研发计划指导类项目（GZ20210110）；黑龙江省中医药管理局中医药经典普及化专项课题项目（ZYW2022-063）；黑龙江省自然科学基金项目（LH2022H001）；哈尔滨商业大学产业化项目支持计划（XL0079）