目的 基于UPLC-MS/MS技术与生物信息学分析方法，结合动物实验探讨舒眠胶囊改善对氯苯丙氨酸（PCPA）致失眠大鼠症状的作用机制。方法 采用UPLC-MS/MS技术鉴定舒眠胶囊的主要化学成分；整合网络药理学与GEO数据库挖掘技术，预测舒眠胶囊治疗失眠的潜在核心靶点与信号通路。通过PCPA诱导失眠大鼠模型，采用戊巴比妥钠睡眠时长实验、旷场实验验证低、中、高剂量（100、200、400 mg·kg^-1）舒眠胶囊的体内治疗效果；通过ELISA法检测大鼠血清中神经递质［5-羟色胺（5-HT）、多巴胺（DA）］、HPA轴相关激素［促肾上腺皮质激素（ACTH）、促肾上腺皮质激素释放激素（CRH）、皮质醇（CORT）］及炎症因子［白细胞介素（IL）-1β、IL-6、肿瘤坏死因子（TNF）-α］水平；采用尼氏（Nissl）染色观察海马区神经元病理形态及尼氏小体变化； Western blotting检测大鼠海马组织中脑源性神经生长因子（BDNF）、酪氨酸激酶受体B（TrkB）、突触后密度蛋白95（PSD95）蛋白表达及环磷腺苷效应元件结合蛋白（CREB）的磷酸化水平。结果 UPLC-MS/MS鉴定出舒眠胶囊中34个化学成分。网络药理学分析显示，舒眠胶囊抗失眠作用主要涉及突触信号传递、膜电位调控及神经递质转运等生物过程。动物实验结果表明，与模型组比较，舒眠胶囊各剂量组能显著延长大鼠睡眠时长，降低自主活动次数；显著提高血清5-HT含量，降低DA、CRH、ACTH、CORT以及IL-1β、IL-6、TNF-α水平；改善海马神经元病理损伤，并显著上调海马组织中BDNF、TrkB、PSD95蛋白表达及CREB磷酸化水平（P＜0.05、0.01）。结论 舒眠胶囊能够有效改善PCPA诱导的大鼠失眠症状，其作用机制与激活BDNF/TrkB/CREB信号通路相关。

Objective Based on UPLC-MS/MS technology and bioinformatics analysis methods, combined with animal experiments, the mechanism of action of Shu Mian Capsules (SMC) in improving insomnia induced by p-chlorophenylalanine (PCPA) in rats was explored. Methods The main chemical components of Shu Mian Capsules were identified using UPLC-MS/MS technology; network pharmacology and GEO database mining techniques were integrated to predict the potential core targets and signaling pathways of Shu Mian Capsules in the treatment of insomnia. Through the PCPA-induced insomnia rat model, pentobarbital sodium sleep duration experiment and open field test were used to verify the in vivo therapeutic effects of Shu Mian Capsules at low, medium, and high doses (100, 200, 400 mg·kg^-1); ELISA was used to detect the levels of neurotransmitters [5-hydroxytryptamine (5-HT), dopamine (DA)], HPA axis-related hormones [adrenocorticotropic hormone (ACTH), corticotropin-releasing hormone (CRH), cortisol (CORT)], and inflammatory factors [interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α] in rat serum; Nissl staining was used to observe the pathological morphology of neurons in the hippocampus and changes in Nissl bodies; Western blotting was used to detect the protein expression of brain-derived neurotrophic factor (BDNF), tyrosine kinase receptor B (TrkB), postsynaptic density protein 95 (PSD95), and the phosphorylation level of cAMP response element binding protein (CREB) in rat hippocampal tissue. Results UPLC-MS/MS identified 34 chemical components in Shu Mian Capsules. Network pharmacology analysis showed that the anti-insomnia effect of Shu Mian Capsules mainly involved biological processes such as synaptic signal transmission, membrane potential regulation, and neurotransmitter transport. Animal experiment results indicated that compared with the model group, Shu Mian Capsules at various doses significantly prolonged the sleep duration of rats and reduced the number of spontaneous activities; significantly increased serum 5-HT levels and decreased DA, CRH, ACTH, CORT, as well as IL-1β, IL-6, and TNF-α levels; improved pathological damage to hippocampal neurons, and significantly upregulated BDNF, TrkB, PSD95 protein expression, and CREB phosphorylation levels in hippocampal tissue (P<0.05, 0.01). Conclusion Shumian Capsules can effectively alleviate insomnia symptoms induced by PCPA in rats, and its mechanism of action is related to the activation of the BDNF/TrkB/CREB signaling pathway.

R965

贵州省科技计划项目（黔科合支撑［2024］一般067，黔科合基础ZD［2025］026，黔科合服企-ZK［2024］005），天产自J字［2025］6）