基于“味性化味-网络靶点-分子对接”的藏药五味麝香丸治疗“真布”病的作用机制研究

Study on the Mechanism of Tibetan Medicine Wuwei Shexiang Pills in the Treatment of“Grum bu”Disease Based on “Ro Nus Zhur Jes-Network Target-Molecular Docking”

目的:探讨藏药五味麝香丸治疗"真布"病(即类风湿性关节炎)的可能机制。方法:以《四部医典》《卫生部药品标准(藏药)》《晶珠本草》为依据,搜集五味麝香丸的组方、剂量及显味,构建矢量结构模型,从六味、三化味、十七效等层面分析该方的药性,并利用Gephi 0.9.2复杂网络软件构建其治疗"真布"病的"组方-药性-疾病"网络。利用中药系统药理学分析平台、有机小分子生物活性数据库检索五味麝香丸的有效成分,并利用BATMAN-TCM网络药理学研究平台预测有效成分对应的靶标蛋白,借助ETCM数据库检索"真布"病的相关靶标蛋白;在筛选两者共同靶标的基础上,利用DAVID 6.8生物信息学资源数据库进行基因本体(GO)分析和KEGG通路富集分析,使用Cytoscape 3.7.0软件构建五味麝香丸-"真布"病-靶标-通路网络并进行网络拓扑学分析,以筛选核心靶标。以Glide score为评价指标,利用Maestro Version 11.1.011软件将上述核心靶标与五味麝香丸有效成分进行分子对接。结果与结论:五味麝香丸含诃子、铁棒锤、木香、藏菖蒲和人工麝香,以诃子剂量最高。该方六味以苦、甘为主,三化味以苦化味为主,十七效以钝、凉、重等为主,主要对治锐、热、轻等二十种特性。"组方-药性-疾病"网络中,十七效与二十种特性之间边权重值较大的为凉效-热性、钝效-锐性、重效-轻性、稀效-臭性(边权重值≥430)等。共得到五味麝香丸有效成分潜在靶标2 306个,"真布"病相关靶标211个;两者共同靶标32个,其对应的有效成分共29种。GO分析共预测到52条相关结果,共同靶标主要位于胞外区、细胞核等部位,以免疫反应、炎症反应、细胞因子活性等生物学过程和分子功能为主。KEGG富集结果显著的有31条(P<0.05),涉及肿瘤坏死因子(TNF)信号通路、癌症途径等信号通路。五味麝香丸-"真布"病-靶标-通路网络中,含有效成分、靶标、通路节点94个、边460条;TNF、自杀相关因子(FAS)、白细胞介素6(IL6)、IL10、自杀相关因子配体超家族成员6(FASLG)、前列腺素内过氧化物合酶2(PTGS2)、IL1B等是该网络中的核心靶标,分别与奎宁酸、百里酚、去氢表雄酮、诺卡酮等有效成分以范德华力、氢键、疏水作用力、Pi-cation键等连接,发挥治疗"真布"病的作用。

OBJECTIVE:To investigate the potential mechanism of Tibetan medicine Wuwei shexiang pills in the treatment of"Grum bu"disease(i.e. rheumatoid arthritis). METHODS:Referring to Sibu Yidian,Drug Standard of the Ministry of Public Health(Tibetan Medicine),Jingzhubencao,the constituent,dose and flavor of Wuwei shexiang pills were collected;vector structure model was built. From the aspects of six tastes,three tastes after digestion and seventeen effects,the properties of the formulation were analyzed;the "formulation-drug property-disease" network for the treatment of "Grum bu" disease was constructed by using Gephi 0.9.2 complex network software. The effective components of Wuwei shexiang pills were screened by using the pharmacology analysis platform of TCM system and the database of organic small molecule biological activity. The target proteins corresponding to the active components were predicted by using BATMAN-TCM network pharmacology research platform;the target proteins of"Grum bu"disease were searched by ETCM database. On the basis of screening the common targets of the two,David 6.8 bioinformatics resource database was adopted to conduct gene ontology(GO) analysis and KEGG pathway enrichment analysis;the network of Wuwei shexiang pills-"Grum bu"-target-pathway was constructed by using Cytoscape 3.7.0 software,and the network topology analysis was carried out to screen the core targets. Using Glide score as evaluation index,Maestro Version 11.1.011 software was used for molecular docking of above core targets with effective components of Wuwei shexiang pills. RESULTS & CONCLUSIONS:Wuwei shexiang pills contains myrobalan,Aconitum pendulum,Aucklandia lappa,Acorus calamus and artificial moschus,and the highest dosage was myrobalan. The six tastes of this formulation were mainly bitter and sweet;the three tastes after digestion were mainly bitter;the seventeen effects were mainly blunt,cool and heavy,which were mainly used to treat twenty characteristics such as acute,hot and light. In"formulation-property-disease"network,higher values of edge weight between seventeen effects and twenty characteristics were cooling effect-heat,blunt effect-sharpness,heavy effectlight,dilute effect-odor(edge weight values≥430),etc. A total of 2 306 potential targets of effective components of Wuwei shexiang pills,211 targets related to"Grum bu"disease,32 common targets and 29 corresponding effective components were obtained. The results of GO analysis showed that 52 relative results were predicted,the common targets mainly located in the extracellular area, nucleus and other parts, mainly including biological processes and molecular functions such as immune response,inflammatory response,cytokine activity. The results of KEGG enrichment were significant in 31 pathways(P<0.05),involving TNF signaling pathway,cancer pathway and other signaling pathways. There were 94 active components,targets and pathway nodes in the network of Wuwei shexiang pills-"Grum bu"-target-pathway,and 460 edges;TNF,FAS,IL6,IL10,FASLG,PTGS2 and IL1 B were the core targets of the network, connected with effective components such as quinic acid,thymol,dehydroepiandrosterone,norcaxone by Van Der Waals force,hydrogen bond and hydrophobic interaction,Pi-cation bond and other bonds,so as to play a role in the treatment of"Grum bu"disease.