昆仑雪菊黄酮类提取物抗脂多糖诱导的小胶质细胞活化作用及网络药理学机制分析  

Flavonoids Extract of Coreopsis tinctoria Nutt.Inhibit Activation of Microglia Induced by LPS and Its Potential Mechanism Based on Network Pharmacology

作　　者：滕李利 钟奕 戚舜尧 孙乐[1,2] 马培[1,2] 许利嘉[1,2] 肖培根 TENG Li-li;ZHONG Yi;QI Shun-yao;SUN Le;MA Pei;XU Li-jia;XIAO Pei-gen(Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100193,China;Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine,Ministry of Education,Beijing 100193,China)

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]中草药物质基础与资源利用教育部重点实验室,北京100193

出　　处：《中国现代中药》2023年第8期1745-1754,共10页Modern Chinese Medicine

基　　金：中国医学科学院医学与健康科技创新工程项目(2022-I2M-2-001);国家自然科学基金项目(82174046);广西科技基地和人才专项(AD22080012)。

摘　　要：目的:研究昆仑雪菊干预小胶质细胞功能的作用机制。方法:采用CCK-8法检测昆仑雪菊总黄酮对BV2细胞的增殖抑制活性,Griess法和ELISA检测细胞炎症因子一氧化氮(NO)、白细胞介素-6(IL-6)的分泌。借助自建昆仑雪菊成分-靶点数据集对昆仑雪菊中9个主要黄酮成分和相关作用靶点进行筛选。通过GeneCards、GENE、STRING数据库及R语言收集并筛选小胶质细胞功能靶点,筛选昆仑雪菊改善小胶质细胞功能的靶点,构建蛋白质-蛋白质相互作用(PPI)网络,并用Cytoscape软件的NetworkAnalyzer功能筛选核心靶点。利用基因集富集分析(GSEA)软件对分子特征数据库MSigDB中的相关靶点进行Hallmark基因分析。使用Bioconductor数据库进行基因本体(GO)功能注释和京都基因与基因组百科全书(KEGG)通路富集分析。结果:昆仑雪菊黄酮类提取物在4μg·mL–1时可以显著抑制小胶质细胞激活分泌NO和IL-6。昆仑雪菊黄酮类成分可能作用于蛋白激酶B(Akt)、肿瘤坏死因子(TNF)、IL-6、TP53、酪氨酸激酶C(SRC)、表皮生长因子受体(EGFR)、JUN、信号转导和转录激活因子3(STAT3)、丝裂原活化蛋白激酶3(MAPK3)、CTNNB1等靶点干预小胶质细胞的功能。涉及的通路包括磷脂酰肌醇3-激酶(PI3K)-Akt信号通路、MAPK信号通路、TNF信号通路、信号转导子和转录激活子(JAK)-STAT信号通路等。结论:昆仑雪菊黄酮类成分可能通过参与调控PI3K-Akt信号通路、MAPK信号通路、TNF信号通路、JAK-STAT信号通路等的相关靶点,实现调节胶质细胞活化的作用。Objective:To study the action mechanism of the intervention of Coreopsis tinctoria Nutt.(CT)on microglia function.Methods:The inhibitory activity of flavonoid extract of CT on BV2 cells was detected by the CCK8 method,and the inflammatory cytokines NO and IL-6 were detected by the Griess and ELISA.The active components and related targets of CT were screened by the self-built component and target data set.Through GeneCards,GENE,STRING database,and R language,the functional targets of microglia were collected and screened,and CT was screened to improve the functional targets of microglia.The protein-protein interaction(PPI)network was constructed.The core nodes were selected by the NetworkAnalyzer function of Cytoscape software.Gene set enrichment analysis(GSEA)software was used for Hallmark gene analysis of related targets in the molecular signature database(MSigDB).Gene ontology(GO)functional annotation and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were carried out by the Bioconductor database.Results:At a dose of 4μg·mL–1,CT significantly inhibited NO and IL-6 activation by microglia.CT may act on Akt,TNF,IL-6,TP53,SRC,EGFR,JUN,STAT3,MAPK3,and CTNNB1 targets,and PI3K-Akt,MAPK,TNF,and JAK-STAT signaling pathways were involved.Conclusion:The flavonoids of CT may regulate the activation of microglia via PI3K-Akt,MAPK,TNF,and JAK-STAT signaling pathways.

关 键 词：昆仑雪菊 小胶质细胞 网络药理学 作用机制 

分 类 号：R285[医药卫生—中药学]