基于网络药理学与体外实验探讨附子活性成分治疗血管痉挛的作用机制  

The mechanism of action of the active components of Aconitum Carmichaelii Debx Rhizoma in the treatment of vasospasm based on network pharmacology and in vitro experiments

作　　者：武博文 颜培宇 武密山 WU Bowen;YAN Peiyu;WU Mishan(Faculty of Chinese Medicine,Macao University of Science and Technology,State Key Laboratory of Quality Research in Chinese Medicines,Macao University of Science and Technology,Macao 999078,China)

机构地区：[1]澳门科技大学中医药学院中药质量研究国家重点实验室,中国澳门999078

出　　处：《环球中医药》2024年第8期1523-1536,共14页Global Traditional Chinese Medicine

基　　金：澳门科技大学研究基金(FRG-22-109-FC)。

摘　　要：目的运用网络药理学和体外细胞实验探究附子活性成分治疗血管痉挛的潜在作用机制。方法检索TCMSP数据库、SwissTargetPrediction数据库、Uniprot数据库、Genecards数据库、OMIM数据库、TTD数据库、Disgenet数据库和String数据库等,获得附子活性成分、作用靶点及血管痉挛相应疾病靶点,并将其作用靶点与获得的疾病靶点的交集作为潜在靶点;利用String平台,对潜在靶点构建蛋白—蛋白互作(protein-protein interaction network,PPI)网络;采用R语言软件包对潜在靶点进行基因本体(gene ontology,GO)功能及京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析,然后使用Cytoscape软件分析其网络拓扑结构,筛选其核心靶点,并进行分子对接验证。在此基础上,通过MTT法检测附子活性成分对血管紧张素Ⅱ所致血管平滑肌细胞增殖影响的机制。结果附子治疗血管痉挛的主要潜在活性成分有15个;药物—疾病核心靶点有27个;涉及的信号通路主要有脂质和动脉粥样硬化、钙信号通路、刺激神经组织的中的交互、鞘脂类信号通路、流体剪切应力与动脉粥样硬化、松弛素信号通路、血管平滑肌收缩等;关键核心靶点与活性成分次乌头碱、谷甾醇结合具有稳定构象。体外实验结果表明,附子活性成分次乌头碱能抑制血管紧张素Ⅱ所致血管平滑肌细胞增殖,其机制可能与其抑制丝裂原活化蛋白激酶信号转导通路的活化有关。结论附子多种活性成分可能通过作用于丝裂原活化蛋白激酶1、核受体亚家族3 C群成员1、血管紧张素I转化酶、一氧化氮合酶3等靶点治疗血管痉挛,为附子活性成分治疗血管痉挛的作用机制研究提供了科学参考。Objective To explore the potential mechanism of the active components of Aconitum Carmichaelii Debx Rhizoma in thetreatment of vasospasm by integrating network pharmacology and in vitro cell experiments.Methods The TCMSP,SwissTargetPrediction,Uniprot,Genecards,OMIM,TTD,Disgenet,String database,and other related literature were searched to obtain active components in Aconitum Carmichaelii Debx Rhizoma,action targets Rhizoma,and corresponding disease targets.The intersection of action and disease targets was taken as the potential targets.A protein-protein interaction network(PPI)for potential targets was constructed based on String platform.The R language software package was used to analyze the function of gene ontology(GO)and the enrichment of Kyoto Encyclopedia of genes and genomes(KEGG),and then Cytoscape software was used to analyze its network topology and further screen core targets.Molecular docking was carried out to verify the predicted results.Subsequently,the effect of active components in Aconitum Carmichaelii Debx Rhizoma on the proliferation of vascular smooth muscle cells(vSMCs)induced by angiotensinⅡ(AngⅡ)was detected by MTT method.Results There are 15 main potential active components of Aconitum Carmichaelii Debx Rhizoma in the treatment of vasospasm and 27 drug-disease targets.The signaling pathways involved mainly include lipid and atherosclerosis,calcium signaling pathway,neuroactive ligand-receptor interaction,sphingolipid signaling pathway,fluid shear stress and atherosclerosis,relaxin signaling pathway,vascular smooth muscle contraction,etc.The stable conformations were formed between the core target and theactive component,hypaconitine and sitosterol.The results in vitro showed that hypaconitine may inhibit AngⅡ-induced proliferation and phenotype transformation of vSMCs through inactivating mitogen-activated protein kinase(MAPK)signaling pathways.Conclusion Various active components of Aconitum Ccarmichaelii Debx Rhizoma may play a role in the treatment of vasospasm by acting on tar

关 键 词：附子 网络药理学 分子对接 血管痉挛 细胞增殖 血管平滑肌细胞 血管紧张素II 

分 类 号：R285[医药卫生—中药学]