基于“质谱分析-网络药理学预测-活性验证”的牛黄解毒丸抗炎作用研究  

Analysis of Anti-Inflammatory Mechanism of Bezoar Antidotal Pill Based on“Mass Spectrometry Analysis-Network Pharmacology Prediction-Activity Verification”

作　　者：伊博文[1] 赵洁[2] 赖华清 唐力英[2] 陈攀龙 柴兴云[3,4] 高小力 吴宏伟[2] YI Bowen;ZHAO Jie;LAI Huaqing;TANG Liying;CHEN Panlong;CHAI Xingyun;GAO Xiaoli;WU Hongwei(Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Chinese Materia Medica, Beijing University of Chinese Medicine,Beijing 102488,China;Center for Modern Research of Chinese Materia Medica,School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100102,China)

机构地区：[1]中国中医科学院西苑医院,北京100091 [2]中国中医科学院中药研究所,北京100700 [3]北京中医药大学中药学院,北京102488 [4]北京中医药大学中药学院中药现代研究中心,北京100102

出　　处：《世界中医药》2022年第7期925-934,共10页World Chinese Medicine

基　　金：国家重点研发计划项目(2018YFC1707302)——中成药处方中“有毒”药味和贵细药精确控制技术研究及其应用。

摘　　要：目的:基于“质谱分析-网络药理学预测-活性验证”整合研究思路,解析牛黄解毒丸抗炎潜在作用机制解析及其活性成分。方法:通过高分辨质谱技术分析牛黄解毒丸化学成分,将分析出的成分;利用中药生物信息学分析工具(BATMAN-TCM)寻找对应的靶标;以“inflammation”作为关键词利用人类孟德尔遗传数据库(OMIM)获得炎症相关靶标;取成分靶标与疾病靶标交集,利用蛋白质-蛋白质相互作用(PPI)数据库(STRING)对共同靶标进行PPI分析,并采用Cytoscape软件对靶标进一步筛选,得到核心靶标;利用生物信息分析工具(DAVID数据库),对核心靶标进行京都基因和基因组百科全书(KEGG)通路富集分析,构建“药物-成分-靶标-通路”网络图;采用AutoDock 4.2.6软件,对关键靶标及相关药物分子进行分子对接验证,并针对前列腺素内过氧化物合酶2(PTGS2)这一靶标与其对应化学成分进行体外实验验证。结果:经高分辨质谱技术分析出牛黄解毒丸中236个成分,对应456个靶标,炎症相关靶标158个,二者交集靶标21个,PPI分析及网络拓扑分析后,获得核心靶标11个,活性成分14个,涉及肿瘤坏死因子(TNF)、核因子κB(NF-κB)、Toll样受体(TLR)和趋化因子等27条通路,分子对接结果显示,PTGS2、白细胞介素-1β(IL-1β)、白细胞介素-13(IL-13)和C-C基序趋化因子配体2(CCL2)等靶标与其对应的大黄酸,虫漆酸D,齐墩果酸等成分结合活性较高,在体外活性实验中,大黄酸对于其对应的PTGS2这一靶标具有较好的抑制作用。结论:本研究构建了牛黄解毒丸抗炎的调控网络,解析了牛黄解毒丸抗炎有效成分和作用机制,发现了大黄酸对PTGS2这一抗炎靶标具有较好的抑制作用。Objective:To analyze the potential mechanism of anti-inflammatory action of Bezoar Antidotal Pill and its active components based on the integrated research idea of“mass spectrometry analysis-network pharmacology prediction-activity verification”.Methods:High-resolution mass spectrometry was used to analyze the components in Bezoar Antidotal Pill,and the analyzed components were used to find the corresponding targets by using the bioinformatics analysis tool of molecular mechanism of traditional Chinese medicine(BATMAN-TCM).Inflammation-related targets were obtained from the Mendelian Inheritance Database(OMIM)database,and the intersection of component targets and disease targets was taken.The protein interaction analysis of common targets was performed using the Protein Interaction Database(STRING)database,and the targets were further screened by Cytoscape software.Using the DAVID database,the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis was performed on the core targets,and a“drug-component-target-pathway”network diagram was constructed;AutoDock 4.2.6 software was used to conduct molecular docking verification on key targets and related drug molecules,and for prostaglandin endoperoxide synthase 2(PTGS2)target and its corresponding chemical constituents for in vitro verification.Results:A total of 236 components in Bezoar Antidotal Pill were analyzed by high-resolution mass spectrometry,corresponding to 456 targets,158 inflammation-related targets,and 21 intersection targets.After protein interaction analysis and network topology analysis,11 core targets and 14 active ingredients were obtained,involving 27 pathways including tumor necrosis factor(TNF),nuclear factor kappa-B(NF-κB),Toll-like receptors(TLR)and chemokines.The docking results showed that the targets of PTGS2,interleukin-1β(IL-1β),interleukin-13(IL-13)and chemokine 2(CCL2)and their corresponding rhubarb Acid,laconic acid D,oleanolic acid and other components had higher binding activities.In vitro activity experi

关 键 词：牛黄解毒丸 抗炎 网络药理学 靶标 

分 类 号：R285[医药卫生—中药学] R284.1[医药卫生—中医学]