基于网络药理学与体外实验探讨川芎活性成分治疗缺血性脑卒中的作用机制  

The Mechanism of Action of the Active Components of Chuanxiong Rhizoma in the Treatment of Ischemic Stroke Based on Network Pharmacology and In Vitro Experiments

作　　者：刘邦 詹锶楷 李克宁[1] 邝杰辉 贤明华 王淑美[1,2,3,4] LIU Bang;ZHAN Sikai;LI Kening;KUANG Jiehui;XIAN Minghua;WANG Shumei(Guangdong Pharmaceutical University,Guangzhou 510006 Guangdong,China;Engineering Technology Research Center for Chinese Materia Medica Quality of Guangdong Province,Guangzhou 510006 Guangdong,China;Key Laboratory of State Administration of TCM for Digital Quality Evaluation Techndogy of Traditional Chinese Medicine,Guangzhou 510006 Guangdong,China;Engineering Technology Research Center for Chinese Materia Medica Ouality of the Universities of Guangdong Province,Guangzhou 510006 Guangdong,China)

机构地区：[1]广东药科大学,广东广州510006 [2]广东省中药质量工程技术研究中心,广东广州510006 [3]国家中医药管理局中药数字化质量评价技术重点研究室,广东广州510006 [4]广东高校中药质量工程技术研究中心,广东广州510006

出　　处：《中药新药与临床药理》2022年第11期1536-1544,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(82004086,81473413)。

摘　　要：目的 运用网络药理学和体外细胞实验探究川芎活性成分治疗缺血性脑卒中的潜在作用机制。方法检索TCMSP数据库、Genecards数据库、Drugbank数据库等及相关文献,获得川芎活性成分、作用靶点及缺血性脑卒中相应疾病靶点,并将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(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CCK-8法检测川芎中的活性成分对缺氧缺糖/再灌注损伤细胞模型的作用,免疫印迹法(Western blot)验证川芎单体成分对相关靶点蛋白的影响。结果 川芎治疗缺血性脑卒中的主要潜在活性成分有16个;药物-疾病靶点有53个;涉及的信号通路主要有神经元神经活性配体-受体相互作用信号通路、钙离子信号通路、血管内皮生长因子信号通路等;关键核心靶点与活性成分藁本内酯[(Z)-Ligustilide]、叶酸(Folic acid)、川芎哚(Perlolyrine)结合具有稳定构象。体外实验结果表明,川芎活性成分藁本内酯能提高缺氧缺糖/再灌注损伤细胞的存活率,并上调转化生长因子β1(TGF-β1)蛋白表达水平。结论 川芎多种活性成分可能通过作用于血管内皮生长因子A(VEGFA)、前列素内环氧化物合成酶2(PTGS2)、雌激素受体(ESR1)、TGFB1等靶点治疗缺血性脑卒中,为川芎活性成分治疗缺血性脑卒中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 Chuanxiong Rhizoma in the treatment of ischemic stroke by integrating network pharmacology and in vitro cell experiments. Methods We searched the TCMSP,Genecards,Drugbank database,and other related literature to obtain active components in Chuanxiong 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 Chuanxiong Rhizoma on oxygenglucose deprivation/reperfusion-injured cell model was detected by CCK-8 method, and the effect of monomer composition in Chuanxiong Rhizoma on related target proteins was verified by Western blot. Results There are 16main potential active components of Chuanxiong Rhizoma in the treatment of ischemic stroke and 53 drug-disease targets. The signaling pathways involved mainly include neuroactive ligand-receptor interaction,calcium signaling pathway, VEGF signaling pathway, etc. The stable conformations were formed between the core target and the active component, such as Z-ligustilide, folic acid, and perlolyrine. The results in vitro showed that ligustilide could improve the survival rate of cells subjected to oxygen-glucose deprivation/reperfusion(ODG/R)and up-regulate the expression level of transforming growth factor β1(TGFβ1) protein. Conclusion Various active components of Chuanxiong Rhizoma may play a role in the treatment of ischemic stroke by acting on targets, such as vascular endothelial growth factor A(VEGFA),prostaglandin epoxide synthase

关 键 词：川芎 分子对接 缺血性脑卒中 网络药理学 缺氧缺糖/再灌注损伤细胞 

分 类 号：R285.5[医药卫生—中药学]