目的:运用网络药理学和生物信息学技术探讨驻春胶囊治疗肾阳虚型骨质疏松症的作用机制.方法:使用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)筛选驻春胶囊组方中药的活性成分,并使用TCMSP和Drugbank数据库检索有效成分相关靶点.在GeneCards数据库、药物靶标数据库(therapeutic target database,TTD)和人类在线孟德尔遗传数据库(online mendelian inheritance in man,OMIM)中检索骨质疏松症致病靶点;在GEO数据库中筛选肾阳虚型骨质疏松症患者与正常人之间的差异表达基因;将两者取交集,得到肾阳虚型骨质疏松症致病靶点.将活性成分相关靶点与肾阳虚型骨质疏松症致病靶点取交集,得到驻春胶囊治疗肾阳虚型骨质疏松症的潜在靶点.利用Cytoscape 3.9.1软件构建"药物-活性成分-潜在靶点"网络.利用R软件对潜在靶点进行基因本体(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopediaof genes and genome,KEGG)信号通路富集分析.采用STRIN G数据库进行聚类关联分析,并通过CytoNCA分析筛选驻春胶囊治疗肾阳虚型骨质疏松症的关键靶基因.采用AutodockVina软件对活性成分和关键靶基因进行分子对接验证;采用CIBERSORT进行免疫细胞浸润分析.结果:本研究共筛选得到183个活性成分、2 732个活性成分相关靶点、897个肾阳虚型骨质疏松症的疾病靶点.将活性成分相关靶点与疾病靶点取交集,得到34个驻春胶囊治疗肾阳虚型骨质疏松症的潜在靶点."药物-活性成分-潜在靶点"网络分析得到木犀草素、8-异戊烯-山柰酚(8-isopentenyl-kaempferol)、黄豆黄素(glycitein)、柚皮素(naringenin)、槲皮素(quercetin)等有效活性成分和SIRT1、CDKN1B、TNFRSF1A等潜在靶基因.GO和KEGG通路富集分析显示,主要靶点富集在多个细胞组分、生物过程及信号通路上.PPI网络分析得到SIRT1、CDKN1B和TNFRSF1A为驻春胶囊治疗肾阳虚型骨质疏松症的关键靶基因.分子对接显示活性成分与关键靶点对接良好.CIBERSORT免疫浸润分析显示,与正常对照组相比,肾阳虚型骨质疏松症组中激活的NK细胞和静止的Mast细胞显著降低,NK细胞的激活与TNFRSF1A的表达呈显著的负相关.结论:驻春胶囊的多个有效活性成分可能通过多个靶点激活多条信号通路发挥改善肾阳虚型骨质疏松症的作用,其中通过TNFRSF1A调控NK细胞的激活可能是驻春胶囊治疗肾阳虚型骨质疏松症的关键环节之一.
Study on Zhuchun Capsules in Treating Kidney-Yang Deficiency Osteoporosis Through TNFRSF1A Regulation of NK Cell Activity
Objective:To explore the mechanism of Zhuchun Capsules in treating kidney-yang deficiency osteoporosis using network phar-macology and bioinformatics techniques.Methods:The active components of Zhuchun Capsules were screened using the Traditional Chi-nese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).Targets related to the active components were identi-fied from the TCMSP and DrugBank databases.Osteoporosis-related targets were retrieved from the GeneCards,Therapeutic Target Data-base(TTD),and Online Mendelian Inheritance in Man(OMIM)database,and differentially expressed genes were identified in kidney-yang deficiency osteoporosis patients from the GEO database.The intersection of active compound-related targets and disease targets was analyzed to identify potential targets for Zhuchun Capsules in treating kidney-yang deficiency osteoporosis.Cytoscape 3.9.1 software was used to construct a"drug-active component-potential target"network.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Ge-nomes(KEGG)pathway enrichment analyses were conducted using R software.STRING and CytoNCA analyses were performed to identify key genes involved in the treatment.Molecular docking was verified using Autodock Vina,and immune infiltration analysis was conducted using CIBERSORT.Results:A total of 183 active components,2 732 related targets and 897 disease-related targets were identified.The intersection revealed 34 potential targets for Zhuchun Capsules in treating kidney-yang deficiency osteoporosis.The"drug-active component-potential target"network analysis identified effective active components such as luteolin,8-isopentenyl-kaempferol,glycitein,naringenin,and quercetin,as well as potential target genes including SIRT1,CDKN1B,and TNFRSF1A,etc.GO and KEGG pathway enrichment analysis revealed that the main targets are enriched in various cellular components,biological processes,and signaling pathways.PPI network analysis identified SIRT1,CDKN1 B,and TNFRSF1A as the key target genes for Zhuchun Capsules in treating kidney-yang deficiency osteoporosis.Molecular docking showed that the active components had good binding affinity with the key targets.CIBERSORT immune infiltration analysis indicated that,compared to the normal control group,the activated NK cells and resting mast cells were significantly reduced in the kidney-yang deficiency osteoporosis group,and NK cell activation was significantly negatively correlated with TNFRSF1A.Conclusion:The multiple active components of Zhuchun Capsules may exert therapeutic effects on kidney-yang deficiency osteoporosis through multiple targets and pathways.The regulation of NK cell activation by TNFRSF1A may be one of the critical steps by which Zhuchun Capsules treat kidney-yang deficiency osteoporosis.
万方数据
维普
