目的 研究醋南柴胡Bupleurum scorzonerifolium抗抑郁潜在的药效物质。方法 采用超高效液相色谱-飞行时间质谱联用（UPLC-Q-TOF-MS）技术，对南柴胡醋炙前后化学成分变化进行全景分析及主成分分析（principal component analysis，PCA），找出差异成分；建立慢性温和不可预测性应激（chronic unpredictable mild stress，CUMS）抑郁大鼠模型，以行为学数据及脑中5-羟色胺（5-hydroxytryptamine，5-HT）、去甲肾上腺素（noradrenaline，NE）及多巴胺（dopamine，DA）含量为评价指标，对醋炙前后的抗抑郁作用进行比较研究；进一步通过网络药理学预测抗抑郁机制及其相关药效物质；建立斑马鱼抑郁模型，检测给药各组斑马鱼行为学数据及5-HT水平，对相关通路进行实验验证。结果 醋南柴胡鉴定出38个成分，其中与生品共有37个。醋炙后13个成分含量发生变化，柴胡皂苷b₁（saikosaponin b₁，SSb₁）、柴胡皂苷b₂（saikosaponin b₂，SSb₂）、秦皮素和矮牵牛素含量升高，柴胡皂苷a（saikosaponin a，SSa）、柴胡皂苷c（saikosaponin c，SSc）、柴胡皂苷d（saikosaponin d，SSd）、异鼠李素-3-O-β-D-葡萄糖苷、异绿原酸A/B/C、柴胡皂苷k（saikosaponin k，SSk）、柴胡皂苷p（saikosaponin p，SSp）、异鼠李素和水仙苷含量降低。醋炙产生了柴胡次皂苷prosaikogenin A/D/G/F。与模型组相比，醋南柴胡组和南柴胡组大鼠体质量、糖水偏好度、旷场实验得分显著增加（P＜0.05、0.01），脑内5-HT、NE、DA含量显著上升（P＜0.05、0.01），醋南柴胡组优于南柴胡组。网络药理学预测IL-17信号通路是抗抑郁关键通路，相关药效物质为SSa、SSd、SSb₁、SSb₂。醋南柴胡及单体皂苷均可提升斑马鱼抑郁模型的运动距离和运动次数，升高体内5-HT含量（P＜0.05，0.01）；逆转录聚合酶链式反应（reverse transcription-polymerase chain reaction，RT-PCR）结果显示均可通过下调白介素-17（interleukin-17，IL-17）、肾素血管紧张素系统（renin-angiotensin system，RAS）、蛋白激酶（raf protein kinase，RAF）、促分裂原活化蛋白激酶激酶（mitogen-activated protein kinase kinase，MEK）、细胞外信号调节激酶（extracellularsignal-regulatedki-nase，ERK）、IL-6、肿瘤坏死因子-α（tumor necrosis factor，TNF-α）mRNA表达水平，调控IL-17信号通路发挥抗抑郁作用。结论 SSa、SSd、SSb₁、SSb₂为醋南柴胡抗抑郁关键药效物质，为阐明南柴胡醋炙增效机制提供了实验依据。

Objective To study the potential antidepressant pharmacodynamic substances of vinegar-baked Bupleurum scorzonerifolium. Methods UPLC-Q-TOF-MS/MS technique was used to qualitatively analyze the chemical constituent change ofB. scorzonerifolium (BS) before and after stir-baking with vinegar, principal component analysis (PCA) was performed to find out differential components. After establishing rat model of chronic mild and unpredictable stress (CUMS), the behavioral data and the contents of 5-hydroxytryptamine (5-HT), noradrenaline (NE), dopamine (DA) in the brain of depressed rats would be the evaluation indexes in order to comparatively study the antidepressant effects among raw and vinegar-baked products; furthermore, the antidepressant mechanism and their associated pharmacodynamic substances were predicted through network pharmacology; establishing a zebrafish depression model to detect the behavioral data and the level of 5-HT in zebrafish, then relevant pathways were verified experimentally. Results There were 38 components of vinegar-baked B. scorzonerifolium (VBS) were identified, 37 of which were common components with raw products. The contents of 13 components changed after vinegar processing, four compounds content increased, including saikosaponin b₁ (SSb₁), SSb₂, fraxetin and petunia, and the contents of saikosaponin a (SSa), saikosaponin c (SSc), saikosaponin d (SSd), isorhanin 3-O-β-D-glucoside, isochlorogenic acid A/B/C, saikosaponin k (SSk), saikosaponin p (SSp), isorhamnetin and narcissus were decreased. In addition, vinegar-producing emerged prosaikogenin A/D/G/F. Compared with model group, the body weight, sugar-water preference, open field test score and intracerebral 5-HT, NE and DA contents of rats in BS and VBS significantly increased (P < 0.05, 0.01), VBS group was superior to the BS group. Network pharmacology predicted that IL-17 signaling pathway was the key pathway to antidepression and SSa, SSd, SSb₁, and SSb₂ were potential antidepressant pharmacodynamic substances. VBS and saikosaponins could improve the distance and rate of the depressed zebrafish and increase 5-HT contents (P < 0.05, 0.01); the result of RT-PCR showed that they exerted antidepressant effects by downregulating expression of IL-17, RAS, RAF, MEK, ERK, IL-6, and TNF-α mRNA, then affecting the IL-17 signaling pathway. Conclusion SSa, SSd, SSb₁, and SSb₂ were the essential antidepressant pharmacodynamic substances of VBS. This study provided the experimental basis for elucidating the synergistic mechanism of BS after stir-baking with vinegar.

R284.1

黑龙江省自然科学基金联合基金培育项目（PL2024H196）