基于网络药理学与分子对接探讨“乳香-没药”治疗膝骨关节炎的作用机制

Analysis of Potential Targets and Mechanisms of“Frankincense-Myrrh”in the Treatment of Knee Osteoarthritis Based on Network Pharmacology and Molecular Docking Techniques

目的:运用网络药理学及高通量分子对接技术的方法,对"乳香-没药"药对治疗膝骨关节炎的潜在靶点及机制进行探讨。方法:利用计算机检索中药系统药理学数据库(TCMSP),分别检索乳香、没药的有效活性成分并筛选得到有效活性成分的靶基因。从OMIM、GeneCards数据库检索膝骨关节炎的相关靶点基因,与"乳香-没药"药对有效成分的靶向基因进行匹配得到交集靶基因,利用Cytoscape 3.7.0软件绘制相关网络图并进行PPI分析。借助R语言程序包对关键基因予以基因本体(GO)功能富集分析以及代谢通路(KEGG)富集分析。利用AutoDock 4.2软件将蛋白互作网络中的核心靶蛋白与关键活性有效成分进行分子对接反向验证。结果:共筛选出"乳香-没药"共计36个活性成分,其中槲皮素、β-谷甾醇、豆甾醇、乳香脂酸、扁枝杉烯、β榄香酮酸为关键有效成分,其干预膝骨关节炎涉及83个交集靶点,并最可能通过作用于白细胞介素-6(IL-6)、Jun激酶(JNK)、白细胞介素-1B(IL-1B)、丝裂原活化蛋白激酶1(MAPK1)等靶点发挥治疗作用。GO分析中生物过程多集中在脂多糖反应、氧化应激反应及细菌来源分子反应等,与膜微结构域、膜区、膜筏等细胞组分密切相关,靶点主要集中在细胞因子活性及细胞因子受体结合等分子功能。KEGG富集分析显示"乳香-没药"干预膝骨关节炎涉及糖尿病并发症信号通路、白细胞介素-17、肿瘤坏死因子、HIF-1等多种信号通路。分子对接结果显示槲皮素与IL-6能稳定地结合并通过氨基酸残基GLU-93(谷氨酸-93)、TYR-97(酪氨酸-97)等发挥相互作用。结论:初步预测了"乳香-没药"治疗膝骨关节炎的靶点为IL-6、JUN、IL1B,涉及炎性、代谢以及肿瘤等多种信号通路,为进一步实验验证提供思路。

Objective:To study the use of network pharmacology and molecular docking techniques,The potential target and mechanism of“Frankincense-Myrrh”in the treatment of knee osteoarthritis were discussed.Methods:The active ingredients of Frankincense and Myrrh were retrieved and the target genes of active ingredients were selected by using the database of pharmacology and analysis platform.Retrieving genes related to knee osteoarthritis from human online Mendelian genetic database,human gene database,Match the target gene of the active ingredient with the“Frankincense-Myrrh”,Draw network diagram using Cytoscape 3.7.0 software.And using protein network interaction analysis,Export the results to the Cytoscape 3.7.0 to visualize the protein interaction network.Gene ontology of key target genes using R language and related data packets And gene pathway enrichment analysis.AutoDock4.2 software was used to verify the molecular docking of the core target protein and the key active components in the protein interaction network.Results:A total of 36 active ingredients of“Frankincense-Myrrh”were selected,Of these,quercetin,β-sterol,soya sterol,frankincolic acid,lentisine,βelemantroketonic acid are the key active components,Its intervention in knee osteoarthritis involves 83 intersection targets,and most likely to play a therapeutic role by acting on targets of IL-6、JUN、IL1 B、MAPK1、PTGS2,GO analysis mainly involves biological processes such as lipopolysaccharide reaction,bacterial molecular reaction,oxidative stress reaction and so on,It is closely related to cell components such as membrane microdomain,membrane region and membrane raft,The target mainly involves the molecular function of cytokine receptor binding,cytokine activity and so on.KEGG enrichment analysis showed that frankincense-myrrh intervention in knee osteoarthritis involved multiple signaling pathways such as diabetic complication signaling pathway,interleukin-17,tumor necrosis factor,HIF-1 and so on.molecular docking results showed that quercetin and IL-6 could bind stably and interact through amino acid residues GLU-93(glutamate-93),TYR-97(tyrosine-97),etc.Conclusion:The target IL-6、JUN、IL1 B,of“Frankincense-Myrrh”in the treatment of knee osteoarthritis is predicted Involved in inflammatory,metabolic and tumor signaling pathways,To provide ideas for further experimental verification.