摘要
目的采用网络药理学方法和分子对接技术探究苦参汤治疗隐孢子虫病的潜在作用靶标及协同作用机制。方法检索中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and AnalysisPlatform, TCMSP)及蛋白质信息数据库(The Universal Protein Resource, Uni Prot),获取苦参汤主要活性成分并预测其潜在靶标。检索中国知网、万方数据、中国生物医学文献数据库(CBM)、Pub Med和Web of Science数据库,对TCMSP数据库未收录的成分加以补充,利用Target Net数据库在线预测所补充活性成分的靶标基因。应用Cytoscape软件进行网络构建与分析,在比较毒物遗传学数据库和Gene Cards数据库中检索隐孢子虫病相关疾病靶标。基于STRING数据库构建靶标蛋白质-蛋白质相互作用网络(PPI),采用DAVID数据库进行基因本体(GO)功能富集分析及京都基因和基因组百科全书(KEGG)通路注释分析,采用Bio GPS数据库定位关键靶标的组织器官分布,应用Auto Dock Tools软件进行分子对接模拟验证。结果共筛选出苦参汤活性成分38种,其中核心成分为槲皮素、(+)-14α-羟基苦参碱、芹菜素等。预测得到831个苦参汤相关靶标及512个隐孢子虫病相关靶标。PPI网络分析发现AKT1、TNF、IL-6等69个关键靶标。苦参汤治疗隐孢子虫病可能与303个生物进程、46种分子功能、29种细胞组分相关,Toll样受体(TLR)、核因子kappa B(NF-?B)、核苷酸结合寡聚化结构域样受体(NLR)信号通路等13条KEGG通路在多靶标协同作用下发挥治疗作用。进一步分析发现,核心靶标主要分布于血液和免疫系统。分子对接显示,活性成分与关键靶标对接的结合能均小于0 k J/mol,配体与受体结合紧密。结论苦参汤中槲皮素、(+)-14α-羟基苦参碱、芹菜素等活性成分可能作用于AKT1、TNF、IL-6等靶标,进而调控TLR、NLR、NF-κB等信号通路,并主要在血液免疫系统中协同发挥对隐孢子虫病的治疗作用。
Objective To explore the potential targets and synergistic mechanisms of Kushen Decoction for the treatment of cryptosporidiosis using network pharmacology and molecular docking methods.Methods The main active ingredients of Kushen Decoction were captured from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and the Universal Protein Resource(UniProt) database, and the potential targets were predicted. In addition, the active ingredients of Kushen Decoction that were not included in the TCMSP database were retrieved in CNKI, WanFang Data, CBM,PubMed and Web of Science databases, and the target genes of all supplemented active ingredients were predicted using the online TargetNet database. Network construction and analysis were performed using the Cytoscape software, and cryptosporidiosisrelated targets were retrieved in the Comparative Toxicogenomics Database and GeneCards database. The protein-protein interaction(PPI) network was created using the STRING database, and the DAVID database was used for GO enrichment and KEGG pathway analyses. The tissue distribution of key targets was investigated using the BioGPS database, and the AutoDockTools software was employed to verify the molecular docking results.Results A total of 38 active ingredients of Kushen Decoction were screened, and the core ingredients included quercetin,(+)-14α-hydroxymatrine and apigenin. A total of 831 targets of Kushen Decoction and 512 cryptosporidiosis-related targets were predicted, and PPI network analysis revealed 69 key targets, including AKT1, TNF and IL-6. There were 303 biological processes, 46 molecular functions and 29 cellular components involved in the treatment of cryptosporidiosis with Kushen Decoction, and 13 KEGG pathways played a therapeutic role in the synergistic mechanisms of multiple targets, such as Toll-like receptor(TLR), nuclear factor kappa B(NF)-?B, nucleotide binding oligomerization domain like receptor(NLR) signal pathways. The core targets were mainly distributed in the hematologic and immune systems. Molecular docking analysis showed that the binding energy between active ingredients and key targets were all less than 0 kJ/mol, indicating the strong binding of ligands to receptors.Conclusions The active ingredients of Kushen Decoction, such as quercetin,(+)-14α-hydroxymatrine and apigenin, may act on targets like AKT1, TNF, IL-6 to modulate TLR, NLR and NF-κB signaling pathways to play a synergistic role in the treatment of cryptosporidiosis in the hematologic and immune system.
作者
詹娜
刘兴海
唐芳莹
张建永
ZHAN Na;LIU Xing-Hai;TANG Fang-Ying;ZHANG Jian-Yong(Zunyi Medical University,Zunyi 563000,China;School of Pharmacy,Zunyi Medical University,China)
出处
《中国血吸虫病防治杂志》
CAS
CSCD
北大核心
2021年第5期483-495,共13页
Chinese Journal of Schistosomiasis Control
基金
遵义医科大学大学生创新创业训练计划项目(ZYDC2020077)。
关键词
隐孢子虫病
苦参汤
网络药理学
分子对接
协同作用机制
Cryptosporidiosis
Kushen Decoction
Network pharmacology
Molecular docking
Synergistic mechanism