中国中药杂志

2020, v.45(08) 1789-1799

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基于网络药理学探讨黄秦艽保肝作用机制
Study on hepatoprotective mechanisms of Veratrilla baillonii based on network pharmacology

何彩静;王晶;梁帅;李竣;黄先菊;
HE Cai-jing;WANG Jing;LIANG Shuai;LI Jun;HUANG Xian-ju;School of Pharmacy, South-Central University for Nationalities;

摘要(Abstract):

该研究旨在预测民族药黄秦艽防治非酒精性肝病作用的活性成分和作用靶点及信号通路,并通过细胞实验对活性成分和相关靶点进行初步验证,阐述黄秦艽保肝的作用机制。通过中药成分及中药药理数据库和分析平台检索,结合药动学参数中的口服利用度和类药性原则筛选黄秦艽候选活性成分,通过蛋白数据库预测候选活性成分靶点,并预测与非酒精性肝病相关的疾病靶点。利用Cytoscape软件构建"活性成分-靶点-疾病"网络,通过STRING数据库构建蛋白互作网络,推测核心靶点。通过DAVID生物信息注释数据库进行GO注释分析、KEGG通路分析以及富集分析。最后通过黄秦艽水提物(WVBF)干预H_2O_2诱导LO2肝细胞损伤模型实验进行核心靶点和通路的初步验证。通过MTT法和实时无标记法检测细胞活力,采用实时定量PCR法分析相关基因的表达量。首先,该文通过网络药理学从黄秦艽中获得14个活性成分,成分对应的潜在靶点共有287个,与非酒精性肝病相关靶点有587个,活性成分靶点与疾病靶点交集后获得13个核心靶点。其次,GO富集分析显示,这些基因主要影响核受体的活性、转录因子的活性、类固醇激素受体活性、泛素样蛋白连接酶结合、蛋白质异二聚化活性、转录辅助因子的结合等26个生物过程。KEGG富集分析显示,PI3K-AKT信号通路、HIF-1信号通路、MAPK信号通路、胰岛素信号通路、TNF信号通路和一些癌症相关通路基因富集较多。最后,通过体外肝细胞实验成功验证TNF-α和MAPK8是其重要靶点,这提示黄秦艽对肝损伤有显著改善作用。由上述结果得出,基于网络预测黄秦艽通过多成分-多靶点-多途径共同调控非酒精性肝病的物质基础和生物学机制,核心靶点通过细胞成功验证,这为民族药黄秦艽的深度开发提供了数据基础和科学依据。
The purpose of this study was to predict the active components, targets and signaling pathways of Veratrilla baillonii for the prevention and treatment of non-alcoholic liver diseases, preliminarily verify the active components and related targets through cell experiments, and elucidate the mechanism of V. baillonii on liver protection. The candidate active components of V. baillonii were screened by searching Chinese medicine ingredients and Chinese medicine pharmacology database and analysis platform, combined with the pharmacokinetic parameters(oral availability and drug-like principle); the target of candidate active ingredients were predicted by protein database, and the target of disease related to non-alcoholic liver disease was predicted. Cytoscape software was used to construct the network of "active component-target-disease", and the protein interaction network was constructed through the STRING database to infer the core target. GO annotation analysis, KEGG pathway analysis and enrichment analysis were conducted through DAVID bioinformatics annotation database. Finally, the core target and pathway of V. baillonii were preliminarily verified by the experimental model of H_2O_2-induced liver cell damage intervened by V. baillonii water extract(WVBF). The cell viability was detected by MTT assay and real-time unlabeled assay, and the expression of related genes was analyzed by Real-time quantitative polymerase chain reaction(PCR). Firstly, 14 active components were obtained from V. baillonii through network pharmacology. There were 287 potential targets corresponding to the components, 587 targets related to non-alcoholic liver disease, and 13 core targets after the interaction between active ingredient targets and disease targets. Secondly, GO enrichment analysis showed that these genes mainly affected 26 biological processes such as nuclear receptor activity, transcription factor activity, steroid hormone receptor activity, ubi-quitin-like protein ligase binding, protein heterodimerization activity, and transcription cofactor binding. KEGG enrichment analysis showed that PI3 K-AKT signaling pathway, HIF-1 signaling pathway, MAPK signaling pathway, insulin signaling pathway, TNF signaling pathway and some cancer-related pathways were more enriched. Finally, TNF-α and MAPK8 were successfully verified as important targets by hepatocytes in vitro, which suggested that V. baillonii could significantly improve liver damage. TNF-α and MAPK8 were one of the targets. Based on the above results, we systematically predicted the material basis and biological mechanism of V. baillonii through multi-component, multi-target and multi-pathway regulation of nonalcoholic liver disease, and the core targets were successfully verified by cells, providing data basis and scientific basis for the in-depth development of V. baillonii.

关键词(KeyWords): 黄秦艽;网络药理学;非酒精性肝病;黄酮类;环烯醚萜;机制;通路
Veratrilla baillonii;network pharmacology;non-alcoholic liver disease;flavonoids;cyclic ethers terpene;mechanism;pathway

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81873090,81374064);; 国家级重大重点项目培育专项(CZP20002)

作者(Author): 何彩静;王晶;梁帅;李竣;黄先菊;
HE Cai-jing;WANG Jing;LIANG Shuai;LI Jun;HUANG Xian-ju;School of Pharmacy, South-Central University for Nationalities;

Email:

DOI: 10.19540/j.cnki.cjcmm.20200115.402

参考文献(References):

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