基于网络药理学和分子对接技术研究黄芩-黄连药对防治2型糖尿病作用机制

Exploration of the mechanism of Scutellariae Radix-Coptidis Rhiʐoma in the prevention and treatment of type 2 diabetes mellitus based on network pharmacology and molecular docking technology

目的 基于网络药理学及分子对接技术探讨黄芩-黄连药对防治2型糖尿病(T2DM)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)收集黄芩-黄连药对的有效成分及对应靶点,借助Perl脚本及UniProt数据库对所得成分及靶点进行分析、整理;从Drugbank数据库、在线人类孟德尔遗传(OMIM)数据库、遗传药理学与药物基因组学数据库(PharmGKB)、疗效药靶数据库(TTD)中收集T2DM相关靶点,绘制韦恩图得药物作用于疾病的交集靶点,并利用Cytoscape 3.8.0软件中的CytoNCA插件筛选核心靶点;对交集靶点进行基因本体(GO)及京都基因与基因组百科全书(KEGG)富集分析;最后使用Vina软件对关键成分和核心靶点进行分子对接。结果 共筛选出黄芩-黄连药对有效成分50个,其中黄连14个,黄芩36个,相关靶点201个,与T2DM 425个靶点取交集得到交集靶点37个;通过“活性成分-靶点”相互作用网络共筛选出关键成分6个(豆甾醇、β-谷甾醇、汉黄芩素、千层纸素、槲皮素、小檗碱),度值前5位的核心靶点为前列腺素内过氧化物合成酶2(PTGS2)、白细胞介素1 B(IL1B)、细胞色素P450 1A1(CYP1A1)、趋势因子白细胞介素-8(CXCL8)、过氧化物酶体增殖物激活受体γ(PPARG);GO富集分析发现黄芩-黄连药对治疗T2DM的基因功能主要富集在血管径调节、肾上腺素能受体的活动蛋白连接、儿茶酚胺结合等过程;KEGG富集分析得到30条信号通路(P<0.05),主要涉及白细胞介素-17(IL-17)信号通路、环磷酸鸟苷-蛋白激酶G(cGMP-PKG)信号通路、脂肪细胞的脂分解调节等。分子对接结果显示,槲皮素、汉黄芩素、千层纸素、豆甾醇与核心靶点PTGS2、CYP1A1、IL1B、PPARG、CXCL8具有良好的结合活性。结论 黄芩-黄连药对主要通过活性成分豆甾醇、β-谷甾醇、汉黄芩素、千层纸素、槲皮素及小檗碱作用于PTGS2、CYP1A1、IL1B、PPARG、CXCL8等靶点,改善血液循环,调节炎症细胞因子、脂肪细胞内脂肪分解,调控cGMP-PKG信号通路以及作用于胰岛素受体起协同效应,从而发挥防治T2DM的作用。

Objective To investigate the mechanism of Scutellariae Radix-Coptidis Rhizoma medicine pair in the prevention and treatment of type 2 diabetes mellitus(T2DM)based on network pharmacology and molecular docking techniques.Methods The active components and corresponding targets of Scutellariae Radix-Coptidis Rhizoma were collected by TCMSP,and analyzed and sorted by Perl script and UniProt database.T2DM related targets were collected from Drugbank,online Human Mendelian Genetics(OMIM),Pharmacogenetics and Pharmacogenomics Knowledge Base(PharmGKB),and Therapeutic Target Database(TTD),and a Venn diagram of the intersection targets of drugs acting on diseases was drawn.The CytoNCA plugin in Cytoscape3.8.0 is used to screen core targets.Gene ontology(GO)and the Kyoto Encyclopedia of Genes and Genomes(KEGG)were analyzed for intersection targets.Finally,Vina software was used for molecular docking of key components and core targets.Results A total of 50 effective ingredients of Scutellariae Radix-Coptidis Rhizoma were selected,14 in Coptidis Rhizoma,36 in Scutellariae Radix with 201 related targets and 37 intersecting targets obtained by intersecting with 425 targets of T2DM.6 key components(stigmasterol,β-sitosterol,baicalin,oroxylin A,quercetin,berberine)were screened out through the“active ingredient-target”interaction network.The top 5 core targets were prostaglandin peroxidase synthetase 2(PTGS2),interleukin1b(IL1B),cytochrome P4501A1(CYP1A1),trend factor interleukin-8(CXCL8),peroxisomal proliferator-activated receptorγ(PPARG).GO enrichment analysis showed that the gene function of Scutellariae Radix-Coptidis Rhizoma on T2DM was mainly concentrated in the process of vascular diameter regulation,active protein linking of adrenergic receptor and catecholamine binding.KEGG enrichment and screening resulted in 30 signaling pathways(P<0.05),mainly involving interleukin-17(IL-17)signaling pathway,cyclic guanosine phosphate protein kinase G(cGMP-PKG)signaling pathway,lipid decomposition regulation of adipocytes,etc.Molecular docking results showed that quercetin,oroxylin A,baicalein,and stigmasterol had good binding activity with core targets PTGS2,CYP1A1,IL1B,PPARG and CXCL8.Conclusion Scutellariae Radix-Coptidis Rhizoma can improve blood circulation,regulate inflammatory cytokines and lipomolysis in adipocytes by targeting PTGS2,CYP1A1,IL1B,PPARG and CXCL8 through active ingredients stigmasterol,β-sitosterol,baicalin,oroxylin A,quercetin,berberine.The regulation of cGMP-PKG signaling pathway and the synergistic effect on insulin receptors play a role in preventing and treating T2DM.