抗痨颗粒治疗结核病分子机制的网络药理和实验验证  

Mechanism of Anti-tuberculosis Particles(Kanglao Granule)Based on Network Pharmacology and Experimental Research

作　　者：肖四方 马小华[1] 石国民 喻容 潘建华 周婷 向延根 XIAO Si-fang;MA Xiao-hua;SHI Guo-min;YU Rong;PAN Jian-hua;ZHOU Ting;XIANG Yan-gen(The Affiliated Changsha Central Hospital Affiliated to University of South China,Changsha 410004,China)

机构地区：[1]南华大学附属长沙中心医院,长沙410004

出　　处：《中国实验方剂学杂志》2022年第4期205-211,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：湖南省自然科学基金项目(2020JJ4639);湖南省卫生健康经济管理科研计划项目(KJZX2021021)。

摘　　要：目的:通过网络药理学探索抗痨颗粒在抗结核分枝杆菌潜在的分子机制。方法:从中药复方数据平台中获得抗痨颗粒的活性成分,通过SwissTargetPrediction获取潜在的靶点,并与GeneCards和美国国家生物技术信息中心(NCBI)数据库筛选出药物治疗的疾病的靶点;采用STRING和Cytoscape 3.8.0构建"中药-疾病靶点-信号通路"网络和筛选关键靶点;然后进行基因本体(GO)和京都基因和基因组百科全书(KEGG)信号通路富集分析;运用AutoDock Vina对抗痨颗粒活性成分和关键蛋白进行分子对接,并通过蛋白免疫印迹法验证活性成分与关键靶蛋白的相互作用。结果:通过筛选去重得到潜在抗痨颗粒重要成分包括β-谷甾醇、芝麻素和山柰酚等化学成分29个;候选作用靶点包括蛋白激酶B1(Akt1),表皮生长因子受体(EGFR),低氧诱导因子-1A(HIF-1A),原癌基因酪氨酸蛋白激酶C(SRC)和基质金属蛋白酶-9(MMP-9)等关键蛋白28个;生物信息分析发现GO得到氧代谢反应、核酸转录和代谢酶途径等41个重要功能条目;KEGG富集分析结核分枝杆菌信号通路、磷脂酰肌醇3-激酶/蛋白激酶B通路等28条重点信号通路;分子对接的结果显示Akt1和芝麻素的结合力最强;体外实验验证芝麻素通过抑制Akt1的磷酸化来达到控制分枝杆菌的生长的作用。结论:抗痨颗粒治疗是通过多组分、多靶点和多途径共同作用间接提高杀菌和免疫反应水平,从而达到治疗结核病的功效,其中Akt1是其参与治疗结核的重要靶蛋白之一。Objective:To explore the potential anti-tuberculosis mechanism of Kanglao granule through network pharmacology.Method:The active components of Kanglao granule were retrieved from related databases and the potential targets of the components from SwissTargetPrediction.Targets of the tuberculosis were screened from GeneCards and National Center for Biotechnology Information(NCBI),and the antituberculosis targets of the prescription were further identified.STRING and Cytoscape 3.8.0 were employed to construct the Chinese medicinal-disease target-signaling pathway network and screen core targets.Then gene ontology(GO)term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were performed.Finally,AutoDock Vina was used for molecular docking between the active components of the prescription and key proteins and Western blotting for verifying the interaction between them.Result:A total of29 important chemical components in the prescription were screened out,includingβ-sitosterol,sesamin,and kaempferol.A total of 28 key anti-tuberculosis targets were retrieved,such as protein kinase B1(Akt1),epidermal growth factor receptor(EGFR),hypoxia inducible factor-1 A(HIF-1 A),proto-oncogene tyrosineprotein kinase(SRC),and matrix metalloproteinase-9(MMP-9).Bioinformatics analysis showed the 28 targets were involved in 41 GO terms such as oxygen metabolism,nucleic acid transcription,and metabolic enzyme pathway,and 28 key KEGG pathways,including Mycobacterium tuberculosis signaling pathway and phosphatidylinositol 3 kinase/protein kinase B pathway.Molecular docking results showed that Akt1 had the strongest binding affinity to sesamin.In vitro experiment indicated that sesamin inhibited the growth of M.tuberculosis by suppressing the phosphorylation of Akt1.Conclusion:Kanglao granule improved the sterilization level and immune response through multi-component,multi-target,and multi-pathway interactions,thereby achieving therapeutic effect on tuberculosis.Akt1 is one of the important targets involved in t

关 键 词：抗痨颗粒 结核病 网络药理学 分子机制 蛋白激酶B 芝麻素 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学]