清热解毒经典药对金银花-连翘干预新型冠状病毒肺炎分子机制及相须配伍实质

Molecular Mechanism of Heat-clearing and Detoxifying Classical Couplet Medicines of Jinyinhua(Lonicerae Japonicae Flos)-Lianqiao(Forsythiae Fructus)in Intervention of COVID-19 and the Key Basis of Mutual Reinforcement between Compatibility

目的运用网络药理学方法探讨清热解毒经典药对金银花-连翘干预新型冠状病毒肺炎(COVID-19)的作用机制,并藉此阐释该药对相须配伍的实质。方法利用中药系统药理学数据库与分析平台(TCMSP)筛选金银花-连翘药对中的活性成分并收集成分相关靶点,将获得的成分靶点与GeneCards数据库中检索到的新型冠状病毒肺炎疾病靶点进行映射,获得金银花-连翘干预COVID-19的作用靶点,利用Cytoscape软件构建“药对-活性成分-干预COVID-19的靶点”网络。得到的靶点利用STRING数据库构建蛋白互作网络并通过CytoHubba筛选关键靶点,在OmicShare在线云平台对靶点进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析,预测其干预COVID-19的作用机制。结果共筛选到32个活性成分,成分相关靶点235个,COVID-19疾病靶点259个,药对-疾病的共同靶点49个。网络拓扑学分析表明,药对干预COVID-19的潜在关键活性成分有槲皮素、木犀草素、山柰酚、β-谷甾醇、汉黄芩素、5-羟基-7-甲氧基-2-(3,4,5)-三甲氧苯基色原酮,其中前4个是药对的共有成分。提取到的关键靶点为TNF、IL6、MAPK1、MAPK8、PTGS2、MAPK14、CXCL8、IL1B、TP53、HMOX1等。GO功能富集分析获得531个生物过程,主要为细胞因子介导的信号通路、细胞对细胞因子刺激的反应、外源性凋亡信号通路负调节、炎性反应等生物学过程;KEGG富集分析获得146条信号通路,主要为癌症通路、糖尿病并发症的晚期糖基化终产物及其受体信号通路、乙型肝炎、肺结核等信号通路。分子对接结果表明,筛选出的6个关键活性成分与SARS-CoV-23CL水解酶有较好的亲和作用。结论金银花-连翘药对通过多个信号通路发挥抗感染、抗炎及免疫调节等作用来干预COVID-19,共有的多个黄酮和植物甾醇类活性成分且对应丰富的COVID-19疾病靶点是金银花-连翘相须配伍成药对的关键。

Objective To explore the mechanism of Jinyinhua(Lonicerae Japonicae Flos)-Lianqiao(Forsythiae Fructus)couplet medicines(JLCM)in intervention of corona virus disease 2019(COVID-19)and elucidate the deep reason of JLCM mutual reinforcement between compatibility by network pharmacology.Methods Active components and targets of JLCM were screened by TCMSP database,TCMSP databases and related literatures were used as supplement,COVID-19 disease targets were screened by GeneGards database,then active components targets and disease targets were mapped.The“couplet medicines-active components-targets related to COVID-19”network was constructed by Cytoscape 3.6.0 software.The protein interaction network map was constructed by STRING database and the core targets were screened out by CytoHubba.Enrichment analysis of GO biological process and KEGG pathway were carried out by OmicSharecloud platform online.Results A total of 32 active components from JLCM correspond to 235 targets.Targets associated with COVID-19 was 259,among them there were 49 mutual targets between JLCM and COVID-19.The“JLCM-active components-target network”topology analysis showed that quercetin,luteolin,kaempferol,beta-sitosterol,wogonin and 5-hydroxy-7-methoxy-2-(3,4,5-trimethoxyphenyl)-chromone are potential key active components,among them the first four components are common ingredients of JLCM.Key targets involved in TNF,IL6,MAPK1,MAPK8,PTGS2,MAPK14,CXCL8,IL1B,TP53,HMOX1.GO function analysis obtained 531 GO biological processes and the targets were concentrated in cytokine-mediated signaling pathway,cellular response to cytokine stimulus,negative regulation of extrinsic apoptotic signaling pathway,inflammatory response and so on.KEGG enrichment analysis obtained 146 KEGG signaling pathways,the targets were rich in pathways in cancer,AGERAGE signaling pathway in diabetic complication,hepatitis B,tuberculosis,IL-17 signaling pathway and other pathways.The results of molecular docking showed that the filtrated key active compounds had good affinity with SARS-COV-23CL hydrolase.Conclusion The JLCM play intervention in COVID-19 by antiinfection,anti-inflammation and immunomodulation through multiple signaling pathways.The several common active components and their abundant targets related to COVID-19 were the key points of JLCM mutual reinforcement between compatibility.