上下两济丹治疗失眠作用机制探析:基于网络药理学、分子对接和实验验证  

Analysis of the Mechanism of Shangxia Liangji Decoction in Treating Insomnia Based on Network Pharmacology,Molecular Docking and Experimental Verification

作　　者：全夏杰 梁豪 纪雄英 蒋莉[2] 张平[2] 欧阳波[4] Quan Xiajie;Liang Hao;Ji Xiongying;Jiang Li;Zhang Ping;Ouyang Bo(Department of Information,Nanhua Hospital Affiliated to University of South China,Hengyang 421002,China;Department of Neurology,Nanhua Hospital Affiliated to University of South China,Hengyang 421002,China;Department of Gastroscopy,Nanhua Hospital Affiliated to University of South China,Hengyang 421002,China;Department of Traditional Chinese Medicine,Nanhua Hospital Affiliated to University of South China,Hengyang 421002,China)

机构地区：[1]南华大学附属南华医院信息科,湖南衡阳421002 [2]南华大学附属南华医院神经内科,湖南衡阳421002 [3]南华大学附属南华医院腔镜室,湖南衡阳421002 [4]南华大学附属南华医院中医科,湖南衡阳421002

出　　处：《亚太传统医药》2024年第11期25-33,共9页Asia-Pacific Traditional Medicine

基　　金：湖南省自然科学基金(2021JJ40510);湖南省临床医疗技术创新引导项目(2021SK51909)。

摘　　要：目的:基于网络药理学和分子对接预测并通过动物实验验证,揭示上下两济丹治疗失眠的潜在分子机制。方法:利用TCMSP数据库获取了上下两济丹的活性成分与对应潜在作用靶点;通过GeneCards和OMIM数据库获取失眠相关的疾病靶点;利用Venny 2.1.0平台对活性成分靶点与疾病靶点取交集,获取上下两济丹治疗失眠的共同靶点;将共同靶点导入STRING数据库,构建蛋白互作(PPI)网络,结合Cytoscape 3.9.0分析后获取关键靶点;并进行GO和KEGG信号通路富集分析;通过分子对接验证核心活性成分与关键靶点的关系;最后建立PCPA诱导的失眠小鼠模型,并给予上下两济丹治疗;RT-qPCR法检测脑组织关键基因的表达。结果:筛选出上下两济丹40个活性成分,预测得到101个活性成分作用靶点,1640个失眠相关靶点;获得24个交集靶点;根据PPI网络的度值获取上下两济丹治疗失眠的5个核心靶点:IL-6、CASP3、VEGFA、PPARG和HIF1A;GO富集分析得到靶点富集于对药物的反应、对脂多糖的反应;KEGG信号通路富集分析得到关键的通路是糖尿病并发症中的AGE-RAGE信号通路、脂质和动脉粥样硬化信号通路;分子对接的结果显示核心成分(Kaempferol、(R)-canadine、Stigmasterol、Beta-sitosterol和Quercetin)能够与关键靶点(IL-6、CASP3、VEGFA、PPARG和HIF1A)紧密结合;戊巴比妥诱导睡眠试验表明,上下两济丹明显缩短睡眠潜伏期,明显延长睡眠持续时间;RT-qPCR显示,上下两济丹治疗降低了IL-6、CASP3、VEGFA、PPARG和HIF1A的mRNA表达水平。结论:上下两济丹可能通过下调IL-6、CASP3、VEGFA、PPARG和HIF1A的表达治疗失眠。Objective:To reveal the potential molecular mechanism of Shangxia Liangji Decoction(SXLJD)in the treatment of insomnia based on the prediction of network pharmacology and molecular docking,and verified by animal experiments.Methods:The active compounds and corresponding potential targets of SXLJD were obtained by TCMSP database.Disease targets related to insomnia were obtained from GeneCards and OMIM databases.The intersection of active ingredient target and disease target was performed by Venny 2.1.0 platform to obtain the common target of SXLJD in treating insomnia.Common targets were imported into STRING database to construct PPI network,and key targets were obtained after combining Cytoscape 3.9.0 analysis.GO and KEGG signaling pathway enrichment analysis was carried out.The relationship between core active ingredients and key targets was verified by molecular docking.Finally,PCPA induced insomnia mouse model was established,and SXLJD's treatment was given.The expression of key genes in brain tissue was detected by RT-qPCR.Results:40 active constituents of SXLJD were screened,101 active compounds and 1640 insomnia related targets were predicted.24 intersection targets were obtained.Five core targets of SXLJD in the treatment of insomnia,IL-6,CASP3,VEGFA,PPARG and HIF1A,were obtained according to the degree value of PPI network.GO enrichment analysis showed that the target was enriched in response to drugs and lipopolysaccharide.KEGG signaling pathway enrichment analysis showed that the key pathways were AGE-RAGE signaling pathway,lipid signaling pathway and atherosclerosis signaling pathway in diabetic complications.Molecular docking results showed that the core compounds(Kaempferol,(R)-canadine,Stigmasterol,Beta-sitosterol and Quercetin)could bind closely to key targets(IL-6,CASP3,VEGFA,PPARG and HIF1A).The pentobarbital induced sleep test showed that SXLJD significantly shortened sleep latency and prolonged sleep duration.RT-qPCR showed that SXLJD's treatment decreased the mRNA expression levels of IL-6,CAS

关 键 词：上下两济丹 失眠 网络药理学 分子对接 

分 类 号：R256.23[医药卫生—中医内科学]