目的 利用网络药理学及分子对接的方法探究养血清脑丸治疗偏头痛的潜在作用机制并对其进行实验验证。方法 利用TCMSP和Swiss Target Prediction数据库筛选养血清脑丸的活性成分和作用靶点，从NCBI GENE、OMIM、CTD、DisGeNET数据库获取与偏头痛密切相关的靶点，将其与养血清脑丸靶点取交集后获得养血清脑丸治疗偏头痛的潜在作用靶点；利用STRING数据库进行蛋白互作网络（PPI）分析，结合Cytoscape 3.6.0软件筛选关键靶点；采用DAVID数据库对养血清脑丸治疗偏头痛的潜在作用靶点进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集分析，并采用Cytoscape 3.6.0软件构建“养血清脑丸–组方中药–活性成分–偏头痛–靶点–通路”网络和筛选关键成分，使用AutoDock软件对关键成分与关键靶点进行分子对接。采用硝酸甘油诱导的偏头痛大鼠模型，对网络药理学研究结果中的部分关键靶点进行实验验证。结果 筛选获得养血清脑丸活性成分140个、作用靶点279个，其中与偏头痛治疗相关的靶点53个；通过网络分析，筛选获得白细胞介素-6（IL-6）、溶质载体家族6成员4（SLC6A4）、肿瘤坏死因子（TNF）、血管内皮生长因子A（VEGFA）、内皮型一氧化氮合酶（NOS3）等靶点、异紫堇杷明碱、槲皮素、斯氏紫堇碱、球紫堇碱、山柰酚等成分可能是养血清脑丸治疗偏头痛的关键靶点和关键成分，分子对接结果表明关键成分和关键靶点间具有较好的结合活性；KEGG通路富集结果表明养血清脑丸可能通过神经活性配体受体相互作用、5-羟色胺能突触、钙信号通路、cAMP信号通路及肿瘤坏死因子信号通路等途径发挥治疗偏头痛的作用。动物实验结果表明，养血清脑丸能抑制偏头痛大鼠血清中一氧化氮（NO）、IL-6、TNF-α及脑组织中VEGF表达的升高，促进脑组织5-羟色胺（5-HT）水平的升高和5-羟色胺受体1B（5-HT1B）蛋白的表达，抑制SLC6A4和CGRP的表达，对硝酸甘油型大鼠偏头痛症状有明显的改善作用。结论 明确了养血清脑丸治疗偏头痛的潜在成分、靶点和通路，其作用机制可能与升高5-HT水平和5-HT1B蛋白表达、抑制SLC6A4和降钙素基因相关肽（CGRP）表达有关，为深入开展其作用机制的研究提供了科学依据和参考。

Objective To explore the potential mechanism of Yangxue Qingnao Pills in treatment of migraine based on network pharmacology and molecular docking and to conduct its experimental verification. Methods TCMSP and Swiss Target Prediction databases were used to screen the active components and targets of Yangxue Qingnao Pills. Genes related to the occurrence and development of migraine were searched through databases of NCBI GENE, OMIM, CTD, and DisGeNET. The potential targets of treatment were obtained by taking the intersection of the two above. STRING database was used for establishing protein-protein interaction (PPI) networks and key targets were selected by Cytoscape 3.6.0. GO and KEGG pathways involved in the targets were analyzed by DAVID database. "Yangxue Qingnao Pills-herbs-active components-migraine-targets-pathways" network was constructed and key components were screened by Cytoscape 3.6.0, AutoDock software was used to dock the molecules of the key components with the key targets. The migraine rat model induced by nitroglycerin was used to verify key nodes in the results of network pharmacology. Results 140 active components and 279 targets of Yangxue Qingnao Pills were screened. 53 targets were identified for the intersection between Yangxue Qingnao Pills and migraine. The key targets of Yangxue Qingnao Pills in treatment of migraine were IL-6, SLC6A4, TNF, VEGFA, NOS3, etc. The key components were isocorypalmine, quercetin, (S)-scoulerine, bulbocapnine, kaempferol, etc. Molecular docking showed a good binding ability of key components and key target. The enrichment analysis of KEGG pathway showed that Yangxue Qingnao Pills treatment for migraine involved multiple pathways, including neuroactive ligand-receptor interaction, serotonergic synapse, calcium signaling pathway, cAMP signaling pathway, TNF signaling pathway, etc. The results of animal experiments showed that Yangxue Qingnao Pills could inhibit NO, IL-6, TNF-α, VEGF, SLC6A4, and CGRP in serum or brain tissue of migraine rats and increase of 5-HT and 5-HT1B level in brain tissue, significantly improve the symptoms of migraine rats. Conclusion This study reveals the potential active components, key targets and related pathways of Yangxue Qingnao Pills in treatment of migraine, the mechanism maybe related to increase of the level of 5-HT and the 5-HT1B protein expression, inhibition of SLC6A4 and CGRP expression, which provides a basis for further experiments.

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山西省教育厅高校科技创新计划项目（2020L0227）；山西省运城市中心医院院级项目（YJ2022107）