文献类型：期刊文献

中文题名：基于网络药理学和分子对接技术探析通痹健骨方治疗膝骨关节炎的作用机制

英文题名：Mechanism of Tongbi Jiangu Prescription in Treatment of Knee Osteoarthritis Based on Network Pharmacology and Molecular Docking

作者：王雷[1];李盛华[2];周明旺[2];王晓萍[2];徐斌[1];魏长浩[1]

第一作者：王雷

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省中医院,兰州730050

第一机构：甘肃中医药大学

年份：2022

卷号：28

期号：3

起止页码：180

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家中医药管理局中医药传承与创新“百千万”人才工程(岐黄工程)岐黄学者(2018)(国中医药人教函2018-284号);国家中医药管理局全国中医学术流派传承工作室建设项目(国中医药人教函2019-62号)。

语种：中文

中文关键词：通痹健骨方;网络药理学;膝骨关节炎;分子对接;作用机制

外文关键词：Tongbi Jiangu prescription;network pharmacology;knee osteoarthritis;molecular docking;mechanism of action

摘要：目的:基于网络药理学和分子对接技术探析通痹健骨方治疗膝骨关节炎(KOA)的作用机制,并进一步运用细胞实验进行验证。方法:依据中药系统药理学数据库与分析平台(TCMSP)筛选出通痹健骨方活性成分及其对应的靶点;通过检索GeneCards数据库,在线人类孟德尔遗传数据库(OMIM)及DrugBank数据库获取KOA靶点;将通痹健骨方活性成分靶点与KOA靶点取交集,所得靶点即为通痹健骨方治疗KOA靶点。采用Cytoscape 3.7.2软件构建活性成分-靶点网络及蛋白质-蛋白质相互作用(PPI)网络;使用STRING数据库进行PPI网络分析;运用DAVID数据分析平台进行基因本体(GO)富集分析及京都基因与基因组百科全书(KEGG)富集分析;选取关键靶点及核心活性成分,使用AutoDock软件进行分子对接。细胞实验验证网络药理学结果并进行药效学观察。结果:网络药理学预测显示通痹健骨方治疗KOA活性成分有111个,核心活性成分为槲皮素,山柰酚,β-谷甾醇,关键靶点为白细胞介素-1β(IL-1β),基质金属蛋白酶-3(MMP-3),肿瘤坏死因子-α(TNF-α)。GO分析中生物过程(BP)主要涉及到炎症反应、对脂多糖的反应、凋亡信号通路和DNA结合转录因子活性的调节等生物过程。细胞组成(CC)则包括了质膜蛋白复合物,RNA聚合酶Ⅱ转录因子复合物,膜筏,丝氨酸/苏氨酸蛋白激酶复合物等方面。分子功能(MF)主要富集在细胞因子受体结合、核受体活性、蛋白质结构域特异性结合、丝氨酸水解酶活性、趋化因子受体结合、一氧化氮合酶调节剂的活性等方面。KEGG分析获取相关信号通路为核转录因子(NF)-κB,两面神激酶(JAK)/信号传导及转录激活蛋白(STAT)及Wnt信号通路;分子对接结果显示,核心活性成分与关键靶点具有较好的结合活性。实验验证结果表明,通痹健骨方能够下调IL-1β,MMP-3,TNF-α,NF-κB p65表达水平(P<0.05),上调NF-κB抑制蛋白(IκB)-α水平(P<0.05)。结论:通痹健骨方治疗KOA的作用机制在于应用槲皮素,山柰酚,β-谷甾醇等活性成分,以IL-1β,MMP-3及TNF-α为关键靶点,通过NF-κB,JAK/STAT及Wnt信号通路发挥治疗膝骨关节炎的作用。
Objective:To explore the mechanism of Tongbi Jiangu prescription(TBJG)in the treatment of knee osteoarthritis(KOA)based on network pharmacology and molecular docking,and further verify it by cell experiments.Method:The active components and the corresponding targets of TBJG were screened out according to the traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP).The targets of KOA were obtained from GeneCards,online mendelian inheritance in man(OMIM),and DrugBank.The common targets of active components of TBJG and KOA were the targets of TBJG against KOA.The active component-target network and protein-protein interaction(PPI)network were constructed by Cytoscape 3.7.2.STRING was used for PPI network analysis.DAVID was used for gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses.Key targets and core active components were selected for molecular docking by AutoDock.The results of network pharmacology were verified by cell experiments and the pharmacodynamic responses were observed.Result:The prediction of network pharmacology showed that there were 111 active components of TBJG in the treatment of KOA.The core active components were quercetin,kaempferol,andβ-sitosterol,and the key targets were interleukin-1β(IL-1β),matrix metalloproteinase-3(MMP-3),and tumor necrosis factor-α(TNF-α).Biological processes(BP)in GO analysis mainly involved inflammatory response,response to lipopolysaccharide,apoptosis signaling pathway,and regulation of DNA activity in binding transcription factor.Cellular components(CC)included plasma membrane protein complex,RNA polymeraseⅡtranscription factor complex,membrane raft,and serine/threonine protein kinase complex.Molecular functions(MF)were mainly enriched in cytokine receptor binding,nuclear receptor activity,protein domain specific binding,serine hydrolase activity,chemokine receptor binding,and activity of nitric oxide synthase regulator.As revealed by the KEGG analysis,the relevant signaling pathways were nuclear factor(NF)-κB,Janus kinase(JAK)/signal transducer and activator of transcription(STAT),and Wnt signaling pathways.Molecular docking results showed that the core active components had good binding activities with key targets.The experimental results showed that TBJG could down-regulate IL-1β,MMP-3,TNF-α,and NF-κB p65 expression levels(P<0.05),and up-regulated NF-κB inhibitor(IκB)-α(P<0.05).Conclusion:The mechanism of TBJG in the treatment of KOA lies in the application of active components such as quercetin,kaempferol,andβ-sitosterol with IL-1β,MMP-3,and TNF-αas key targets through the NF-κB,JAK/STAT,and Wnt signaling pathways.