基于网络药理学和分子对接探讨大黄治疗脓毒症的作用机制

Mechanism of Rhei Radix et Rhizoma in treatment of sepsis based on network pharmacology and molecular docking

目的基于网络药理学及分子对接技术探究大黄治疗脓毒症的作用机制。方法通过中药系统药理学数据库与分析平台(TCMSP)获取大黄的有效活性成分及其作用靶点;应用Cytoscape 3.9.1构建大黄–活性成分–靶点的网络图;检索GeneCards、OMIM、Drugbank、TTD数据库,得到脓毒症相关靶点;通过建立维恩图获得中药与疾病交叉的关键靶点;应用STRING平台构建关键靶点蛋白质相互作用(PPI)网络图;使用DAVID数据库对关键靶点进行基因本体(GO)生物学富集分析及京都基因与基因组百科全书(KEGG)通路富集分析。最后,应用PyMOL软件和AutoDock Vina软件对大黄活性成分及其关键靶点进行分子对接验证。结果获得大黄10种有效活性成分,其中重要的成分包括β-谷甾醇、芦荟大黄素、泽兰黄醇素等;药物靶点与疾病靶点交集后获得34个关键靶点,核心靶点有肿瘤坏死因子(TNF)、白细胞介素-1B(IL-1β)、肿瘤蛋白p53(TP53)、原癌基因(MYC)、半胱氨酸天冬氨酸蛋白酶-3(CASP3)、前列腺素内过氧化物合酶2(PTGS2)、过氧化物酶体增殖物激活受体G(PPARG)、转录因子AP-1(JUN)、雌激素受体1(ESR1)、半胱氨酸蛋白酶8(CASP8)。GO富集分析表明大黄的关键靶点主要富集于306个生物过程、29个细胞组成和61个分子功能;KEGG通路富集分析确定了111条相关信号通路,包括磷脂酰肌醇-3-羟激酶(PI3K)-蛋白激酶B(Akt)信号通路、丝裂原活化蛋白激酶(MAPK)信号通路、糖尿病并发症相关的晚期糖基化终末化产物(AGE)-晚期糖基化终末产物受体(RAGE)信号通路等;分子对接结果显示,大黄的主要活性成分芦荟大黄素与关键蛋白TNF、IL-1β、TP53、MYC、CASP3拥有良好的结合能力。结论大黄通过多成分、多靶点和多通路治疗脓毒症。

Objective To explore the therapeutic effect and mechanism of Rhei Radix et Rhizoma on sepsis based on network pharmacology and molecular docking.Methods Active components and action targets of Rhei Radix et Rhizoma were obtained by using TCMSP database.The target network diagram“drug–component-target”of Rhei Radix et Rhizoma in treating sepsis was constructed by using Cytoscape 3.9.1.The sepsis disease targets were screened with GeneCards,OMIM,Drugbank,and TTD databases.The intersection gene targets were found by establishing Venn map of drug target genes and disease targets.The intersection genes were imported into STRING database to construct a protein-protein interaction(PPI)network.The genetic ontology(GO)biological process and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were carried out through DAVID database.Finally,molecular docking technology was used to verify the result between ingredients and targets.Results A total of 10 active components of Rhei Radix et Rhizoma were obtained by TCMSP,includingβ-sitosterol,aloe-emodin,and eupatin.The number of targets that intersect with sepsis was 34.TNF,IL-1B,TP53,MYC,CASP3,PTGS2,PPARG,JUN,ESR1,CASP8 may be the key targets of rhubarb in treatment of sepsis.GO enrichment analysis showed that the key targets of Rhei Radix et Rhizoma were mainly enriched in 306 biological processes,29 cell components and 61 molecular functions.KEGG pathway enrichment analysis identified 111 related signaling pathways,including PI3K-Akt signaling pathway,MAPK signaling pathway,and AGE-RAGE signaling pathway in diabetic complications.The results of molecular docking showed that aloe-emodin,the main active component of Rhei Radix et Rhizoma,had good binding ability with the key proteins TNF,IL-1β,TP53,MYC,and CASP3.Conclusions Rhei Radix et Rhizoma treats sepsis through multi-component,multi-target and multi-pathway treatment.