杜仲水提物改善动物睡眠障碍作用机制的网络药理学分析及实验动物验证

Network Pharmacology Analysis and Experimental Verification of the Mechanism Underly the Improvement Effect of Eucommia ulmoides Oliver Water Extract on the Sleep Disorders in Animals

利用网络药理学分析方法,经过动物试验验证,探究杜仲改善小鼠睡眠障碍的作用机制。通过TCMSP数据库获得杜仲的潜在作用靶点,利用OMIM和GeneCards数据库获得睡眠障碍潜在靶点。将两个靶点合集交集后构建蛋白互作网络,筛选核心靶点。通过David在线平台进行基因本体功能(GO)富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。使用Cytoscape软件构建杜仲成分-睡眠障碍靶点-通路网络,之后采用分子对接技术进行检验。最终通过PCPA小鼠睡眠障碍模型进行体内试验验证。网络药理学和分子对接结果显示,杜仲有效成分烟碱单宁、表奎宁定等通过调节多巴胺D2受体基因(DRD2)和GABAA受体α亚基基因(GABRA1)的表达,发挥镇静安神、抗焦虑的作用。动物试验结果显示,杜仲显著减少了睡眠障碍小鼠的运动量(P<0.05),下调睡眠障碍小鼠下丘脑组织中DRD2基因的表达(P<0.01),上调GABRA1基因的表达(P<0.01)。预测并验证了杜仲改善睡眠障碍的作用机制,为其临床应用提供了一定的参考依据。

The present study investigated the action mechanism of Eucommia ulmoides Oliver in improving sleep disorder by using network pharmacology and animal experiments.The TCMSP database was used to obtain the potential targets of Eucommia ulmoides Oliver.The OMIM and GeneCards databases were used to obtain the potential targets of sleep disorders.Then the intersection was taken to construct the protein interaction network to screen the core targets.Gene ontology(GO)analysis and enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes(KEGG)were performed by using the DAVID database.The Eucommia constituent-sleep disorder target-pathway network was constructed by using Cytoscape software.The molecular docking technique was used to examine it.Finally,the mouse sleep disorder model induced by PCPA was adopted for in vivo experimental validation.The results of network pharmacology and molecular docking showed that the active ingredients of Eucommia ulmoides Oliver,such as(-)-tabernemontanine and epiquinidine,exert sedative and anxiolytic effects by regulating the expression of dopamine D2 receptor(DRD2)and GABAA receptor alpha subunit(GABRA1)genes.The results of in vivo experiments showed that Eucommia ulmoides Oliver significantly reduced the exercise of sleep-disordered mice(P<0.05),down-regulated the expression of DRD2 gene(P<0.01)and up-regulated the expression of GABRA1 gene(P<0.01)in the hypothalamus tissue of sleep-disordered mice.This study predicted and validated the action mechanism of Eucommia ulmoides Oliver in improving sleep disorders and provided some reference for its clinical application.