基于网络药理学探析五味子防治稽留流产的机制及验证  

Mechanism of Schisandrae Chinensis Fructus on Treating Missed Abortion Based on Network Pharmacology and Preliminary Verification

作　　者：梁婧[1] 乔林静 侯海燕 LIANG Jing;QIAO Lin-jing;HOU Hai-yan(Guang′anmen Hospital South Campus,China Academy of Chinese Medical Sciences,Beijing 102618,China;Guang′anmen Hospital(Baoding),China Academy of Chinese Medical Sciences,Baoding 071000,Hebei Province,China;The Second Hospital of Tianjin Medical University,Tianjin 300211,China)

机构地区：[1]中国中医科学院广安门医院(南区),北京102618 [2]中国中医科学院广安门医院保定医院 [3]天津医科大学第二医院

出　　处：《国际妇产科学杂志》2024年第2期167-175,共9页Journal of International Obstetrics and Gynecology

基　　金：中国中医科学院科技创新工程--重大攻关项目妇科学学科资助项目(CI2021A02405);国家自然科学基金青年项目(82104921);中国中医科学院广安门医院南区院级科研基金课题(Y2019-14)。

摘　　要：目的:基于网络药理学方法分析五味子防治稽留流产的潜在作用机制,并通过实验进行验证。方法:运用系统药理学方法查找并从中药系统药理学数据库和分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP,https://old.tcmsp-e.com/tcmsp.php)筛选五味子的活性成分及靶点,查找稽留流产相关基因,确定五味子防治稽留流产的靶点。利用Cytoscape构建“药物成分-靶点”网络,筛选关键化合物。利用String建立蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,并通过Cytoscape-CytoNCA拓扑分析筛选核心靶点。通过对靶基因进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)分析,以预测其可能的信号通路及机制。最后借助人绒毛膜外滋养层细胞系(HTR-8/SVneo)利用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,qRT-PCR)检测和蛋白质印迹(Western blotting)检测进行机制验证。结果:从TCMSP数据库中鉴定了7种五味子活性成分。PPI网络表明雄激素受体(androgen receptor,AR)、雌激素受体(estrogen receptor,ER)、环氧合酶-2(cyclooxygenase-2,COX-2)、核受体辅激活因子2(nuclear receptor coactivator 2,NCOA2)和乙酰胆碱酯酶(acetylcholine esterase,ACHE)可能是五味子防治稽留流产的核心作用靶点。GO富集分析获得44个细胞生物学过程,KEGG途径富集分析获得2个相关信号通路,主要包括甲状腺激素信号通路和雌激素信号通路;在利用细胞模型验证机制过程中发现,二羟环氧苯并芘[benzo(a)pyren-7,8-dihydrodiol-9,10-epoxide,BPDE]染毒后的细胞中AR、ER和COX-2的蛋白质和(或)mRNA水平高于正常细胞,而先使用五味子乙素(Schisandrin B,Sch B)预处理后再染毒BPDE的细胞中AR、ER和COX-2的蛋白质和(或)mRNA水平均较未用Sch B预处理的细胞降低,而NCOA2和ACHE的蛋白质和(�Objective:To explore the potential mechanism of Schisandrae chinensis fructus on the treatment of missed abortion based on a network pharmacology approach and verified experimentally.Methods:The system pharmacology approach was used to find and screen the active components and targets of Schisandrae chinensis from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP,https://old.tcmsp-e.com/tcmsp.php),find the genes related to missed abortion,and identify the targets of Schisandra chinensis for prevention and treatment of missed abortion.The"drug component-target"network was constructed by using Cytoscape to screen key compounds.The protein-protein interaction(PPI)network was established by String,and the core targets were screened by the Cytoscape-CytoNCA topology analysis.Gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG)analysis were performed on the target genes to predict their possible signal pathways and explore their molecular mechanisms.Finally,the mechanism was validated with the help of human chorionic villous trophoblast cell line(HTR-8/SVneo)using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)detection and Western blotting assays.Results:Seven active compounds of Schisandra chinensis were identified from TCMSP database.Schisandrae chinensis has five potential targets for the prevention and treatment of missed abortion.The PPI network shows that androgen receptor(AR),estrogen receptor(ER),cyclooxygenase-2(COX-2),nuclear receptor coactivator 2(NCOA2)and acetylcholine esterase(ACHE)may be the core targets of Schisandrae chinensis for the prevention and treatment of missed abortion.Forty-four cell biological processes were obtained by GO enrichment analysis,and two related signal pathways were obtained by KEGG pathway enrichment analysis,mainly including thyroid hormone signal pathway and estrogen signal pathway.During the process of validating the mechanism using a cell model,it was found that the protein and/or mRNA levels of

关 键 词：五味子 流产 稽留 机制 网络药理学 

分 类 号：R285[医药卫生—中药学]