中药药理与临床

目的:基于网络药理学预测白芷治疗偏头痛的主要成分及相关靶点通路，并考察白芷对实验性偏头痛大鼠的作用及机制。方法:在中药系统药理学数据库(TCMSP)平台收集白芷的活性成分及其对应的靶点，检索GeneCards、DrugBank数据库得到偏头痛相关靶点，将白芷活性成分靶点与偏头痛靶点取交集，采用Cytoscape软件构建“药物-成分-疾病-靶点”网络；通过String数据库构建蛋白相互作用网络。通过DAVID平台，进行基因功能(GO)分析和京都基因与基因组百科全书(KEGG)通路富集分析；选取关键靶点及核心活性成分，使用AutoDock软件进行分子对接。用硝酸甘油建立偏头痛大鼠模型，设置正常对照组、模型对照组、盐酸氟桂利嗪0.001 g/kg组、白芷水煎液0.83、3.33 g/kg组，应用酶联免疫吸附法检测大鼠血清中降钙素基因相关肽(CGRP)、5-羟色胺(5-HT)、一氧化氮(NO)、内皮素(ET)和脑组织中5-羟色胺(5-HT)、多巴胺(DA)、去甲肾上腺素(NE)的含量；通过上述指标变化评价白芷对偏头痛大鼠的干预作用。结果:网络药理学筛选得到18个白芷对偏头痛有效的活性成分，其中β-谷甾醇、豆甾醇、别欧前胡素、珊瑚菜素、异珊瑚菜素这5种为治疗偏头痛的核心成分；筛选出31个共有靶点，其中雌激素受体(ESR1)、半胱氨酸蛋白酶(CASP3、CASP8)、前列腺素内过氧化物酶(PTGS2)、即早表达基因(JUN)为潜在药效靶点；通路注释分析得到38条信号通路，其中神经活性配体-受体相互作用、5-羟色胺能突触信号通路等20条与偏头痛有关。将上述5种成分与5个靶点进行分子对接，其亲和力均小于-3 kcal/mol。动物实验结果显示，白芷能显著降低偏头痛模型大鼠血清中CGRP、5-HT、NO、ET和脑组织中5-HT、DA、NE含量。结论:白芷对大鼠的偏头痛症状有显著改善作用，其主要有效成分可能为植物甾醇及珊瑚菜素、异珊瑚菜素等香豆素类成分；其作用机制可能与作用于5-HT受体，影响5-羟色胺能突触等信号通路，进而降低5-HT等递质的含量有关。

Objective: To predict the main components, targets, and pathways of Baizhi(白芷) against experimental migraine in rats based on network pharmacology, and investigate the effect of Baizhi on experimental migraine rats. Methods: Active components of Baizhi and targets of the components were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and migraine-related targets from GeneCards and DrugBank. The common targets of the active components of Baizhi and migraine were summarized and the "medicinal-active component-disease-target" network was constructed by Cytoscape. String was used to construct protein-protein interaction network, and DAVID was employed for Gene Ontology(GO) term and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment. Key targets and core active components were selected for molecular docking by AutoDock. Migraine was induced in rats with nitroglycerin. Rats were classified into normal group, model group, 0.001 g/kg flunarizine hydrochloride group, and Baizhi decoction groups(3.33 g/kg, 0.83 g/kg). The content of calcitonin gene-related peptide(CGRP), 5-hydroxytryptamine(5-HT), nitric oxide(NO), and endothelin(ET) in serum and 5-HT, dopamine(DA), and norepinephrine(NE) in brain tissues was detected by enzyme-linked immunosorbent assay(ELISA). The effect of Baizhi on migraine rats was evaluated based the changes of the above indicators. Results: A total of 18 active components of Baizhi against migraine were screened out by network pharmacology, among which β-sitosterol, stigmasterol, prangenidin, phellopterin, and cnidilin were the key components. The medicinal and the disease shared 31 common targets, among which estrogen receptor 1(ESR1), cysteine proteinases(CASP3, CASP8), prostaglandin-endoperoxide synthase 2(PTGS2), and JUN were potential drug targets. Among the 38 KEGG pathways, 20 were associated with migraine, such as neuroactive ligand-receptor interaction and 5-hydroxytryptaminergic synaptic signal pathways. The molecular docking of the above five components with five targets showed that their affinity was less than-3 kcal/mol. In the animal experiment, the content of CGRP, 5-HT, NO, and ET in the serum and content of 5-HT, DA, and NE in brain tissue of migraine rats were significantly reduced by Baizhi. Conclusion: The migraine symptoms in rats were significantly alleviated by Baizhi. The main effective ingredients may be phytosterol, phellopterin, cnidilin, and other coumarins. The mechanism is that it can act on 5-HT receptor, affecting 5-hydroxytryptaminergic synaptic signal pathways and down regulating the content of 5-HT and other neurotransmitters.