溶髓方干预终板软骨细胞凋亡作用机制研究:基于网络药理学与分子对接技术

Network Pharmacology and Molecular Docking Based Study on the Mechanism of Action of Myelinolytic Formula Intervening in Endplate Chondrocyte Apoptosis

目的:应用网络药理学和分子对接技术,探讨溶髓方干预终板软骨细胞凋亡(CA)的潜在作用机制,为治疗颈椎间盘退行性病变(CIDD)寻找新思路。方法:通过中药系统药理学分析平台(TCMSP)和中医药百科全书(ETCM),获取溶髓方中13种药物的活性成分和预测分子潜在作用靶点。检索Gennecards和OMIM数据库,筛选出与CA进程相关的靶基因,并与药物靶点取交集。应用Cytoscasp 3.7.2软件构建“中药-活性成分-靶点”网络并借助CytoNCA绘制核心靶点拓扑网络进行拓扑分析,应用STRING数据库构建蛋白互作网络并筛选出核心靶点,使用R4.2.6软件对差异基因进行GO功能富集分析、KEGG通路富集分析。通过AutoDock4和Python3.2对获得的核心成分和核心靶点进行分子对接。结果:(1)获得溶髓方治疗CA的154个活性成分,主要为槲皮素、γ-谷甾醇、山柰酚、黄芩素、桑辛素C等,通过拓扑网络筛选出关键候选基因肿瘤蛋白P53(TP53)、丝裂原活化蛋白激酶3(MAPK3)、Jun原癌基因(JunProto-Oncogene,JUN)等。KEGG富集分析显示,与CA治疗相关的信号通路主要为AGE-RAGE信号通路、IL-17信号通路、TNF信号通路、NF-κB信号通路;(2)分子对接结果显示,溶髓方中核心有效成分与CA核心靶点均能通过氢键结合,且分子对接结合能均≤-5.0 kcal/mol,显现出良好的结合力。结论:溶髓方干预CA进程具有多成分、多靶点、多途径等特征,作用机制可能是溶髓方中的槲皮素、γ-谷甾醇、山柰酚、黄芩素、桑辛素C等活性成分作用于TP53、MAPK3、JUN等靶点,通过调控AGE-RAGE信号通路、IL-17信号通路、TNF信号通路、NF-κB信号通路等信号通路发挥干预CA进程的作用。

Objective:Using network pharmacology and molecular docking techniques,we explored the potential mechanism of action of Rongsui recipe to interfere with chondrocyte apoptosis(CA) in the endplate,and found new ideas for the treatment of cervical intervertebral disc degeneration(CIDD).Methods:Through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and the Encyclopedia of Chinese Medicine(ETCM),the active ingredients and predicted targets of 13 drugs in the Rongsui recipe were obtained.The Gennecards and OMIM databases were searched to screen for target genes associated with CA processes and intersected with drug targets.Cytoscasp 3.7.2 software was used to construct the “drug-active ingredient-target” network and map the core target topology network with the help of CytoNCA for topology analysis,and STRING database was used to construct the protein interaction network and screen the core targets.GO functional enrichment analysis,KEGG pathway enrichment analysis.The obtained core components and core targets were molecularly docked by AutoDock 4 and Python 3.2.Results:The 154 active ingredients of the Rongsui recipe for CA treatment were obtained,mainly quercetin,gamma-sitosterol,kaempferol,baicalein and moracin C.The topological network screened key candidate genes as tumor protein P53(TP53),mitogen-activated protein kinase 3(MAPK3),JunProto-Oncogene(JUN),etc.KEGG enrichment analysis showed that the signaling pathways associated with CA treatment were mainly AGE-RAGE signaling pathway,IL-17 signaling pathway,TNF signaling pathway,and NF-κB signaling pathway.The molecular docking results showed that the core active ingredients in the Rongsui recipe could all bind to the core targets of CA by hydrogen bonding,and the molecular docking binding energies were all≤-5.0 kcal/mol,showing good binding power.Conclusion:The mechanism of action may be that quercetin,gamma-sitosterol,kaempferol,baicalein,moracin C and other active ingredients in Rongsui recipe act on TP53,MAPK3,JUN and other targets to regulate AGE-RAGE signaling pathway,IL-17 signaling pathway,TNF signaling pathway,NF-κB signaling pathway and other signaling pathways to interfere with the CA process.