基于网络药理学与分子对接的腰痛宁胶囊治疗腰椎间盘突出症的作用机制研究

The Mechanism of Yaotongning Capsule(腰痛宁胶囊)in Treating Lumbar Disc Herniation:based on Net⁃work Pharmacology and Molecular Docking

目的探讨腰痛宁胶囊治疗腰椎间盘突出症(LDH)的作用机制。方法采用网络药理学方法,借助中药系统药理学数据库与分析平台(TCMSP)、中医药综合数据库(TCMID)、Genecards数据库与DisGeNET数据库筛选出腰痛宁胶囊有效活性成分及其作用靶点,获取LDH相关疾病靶点;绘制韦恩图并取得腰痛宁胶囊治疗LDH的潜在靶点后,使用STRING 11.5平台构建腰痛宁胶囊治疗LDH的蛋白质-蛋白质相互作用(PPI)网络,并使用Cytoscape 3.7.2软件识别其主要功能子簇及其上的核心靶点;通过生物学信息注释数据库(DAVID)与京都基因与基因组百科全书(KEGG)对核心靶点进行基因本体论(GO)与KEGG富集分析,通过Cytoscape 3.7.2软件构建腰痛宁胶囊治疗LDH的“药物-成分-靶点”网络,运用Autodock4及Pymol 2.2.0软件进行分子对接验证。结果共筛选获得腰痛宁胶囊活性成分537个,作用靶点1335个,LDH相关疾病靶点812个,绘制韦恩图取交集获得潜在靶点155个;构建PPI网络后识别出主要功能子簇5个,包含43个核心靶点;GO富集分析显示核心靶点共涉及346个生物过程、30个细胞组成以及65个分子功能;KEGG富集分析显示核心靶点共涉及252条通路,并与其中磷脂酰肌醇3激酶-蛋白激酶B(PI3K-Akt)信号通路相关性较强;“药物-成分-靶点”网络显示腰痛宁胶囊共包含129种与治疗LDH直接相关的活性成分,分子对接验证显示,关键靶点白细胞介素8(IL8)、基质金属蛋白酶1(MMP1)、基质金属蛋白酶9(MMP9)与活性成分汉黄芩素、木犀草素、山柰酚、槲皮素结合能<-5 kcal/mol,显示出良好的结合能力。结论腰痛宁胶囊治疗LDH的作用机制可能是通过汉黄芩素、木犀草素、山柰酚、槲皮素与IL8、MMP1、MMP9三个靶点的结合,抑制PI3K-Akt信号通路从而减轻机械痛阈值并缓解神经病理性疼痛实现。

Objective To explore the mechanism of Yaotongning Capsule(腰痛宁胶囊)in treating lumbar disc herniation(LDH).Methods Traditional Chinese medicine(TCM)systematic pharmacology database and analysis platform(TCMSP),TCM comprehensive database(TCMID),Genecards database and DisGeNET database were used to screen out the active ingredients and their target points of Yaotongning Capsule through network pharmacology,and LDH-related disease targets were obtained.After drawing Wayne diagram and obtaining the potential targets of Yaotongning Capsule for LDH,the protein-protein interaction(PPI)network of Yaotongning Capsule for LDH was constructed using STRING 11.5 platform,and the main functional sub-clusters and their core targets were identified by Cytoscape 3.7.2 software.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed for core targets based on the annotated database of biological information(DAVID)and the KEGG,while the“drug-ingredient-target”network of Yaotongning Capsule for LDH treatment was constructed by Cytoscape 3.7.2 software,and molecular docking verification was carried out by Autodock 4 and Pymol software.Results A total of 537 active ingredients of Yaotongning Capsule,1335 target points of action and 812 target points of LDH-related diseases were screened out,and 155 potential targets were obtained by drawing Wayne diagram and taking the intersection.After constructing PPI network,five main functional sub-clusters were identified,including 43 core targets.GO enrichment analysis showed that the core targets were involved with 346 biological processes,30 cell components and 65 molecular functions.KEGG enrichment analysis showed that the core targets were involved with a total of 252 pathways,and were closely realted to PI3K-Akt signaling pathway.The“drug-ingredient-target”network showed that Yaotongning Capsule contained 129 active ingredients directly related to LDH treatment.Molecular docking verification showed that the binding energy of key targets interleukin-8(IL8),matrix metalloproteinase-1(MMP1)and matrix metalloproteinase-9(MMP9)with active ingredients wogonin,luteolin,kaempferol and quercetin≤-5 kcal/mol,which showed good binding ability.Conclusion Yaotongning Capsule may treat LDH through the combination of wogonin,luteolin,kaempferol and quercetin with IL8,MMP1 and MMP9 targets,which can inhibit PI3K-Akt signaling pathway and thereby lowering the mechanical pain threshold and relieving neuropathic pain.