摘要
目的:采用网络药理学方法和分子对接技术并结合体内实验,分析并验证补肾启智方调控海马神经元铁死亡改善血管性痴呆(vascular dementia,VD)认知功能的作用机制。方法:在TCMSP数据库、BATMAN-TCM数据库、ETCM数据库筛选补肾启智方的活性成分及其作用靶点;在GeneCards数据库、OMIM数据库、DisGeNET数据库筛选血管性痴呆的潜在靶点;在FerrDB数据库筛选铁死亡的相关靶点,以上所有靶点均经Uniprot数据库规范,并在微生信-免费在线绘制韦恩图——Venn Diagrams网站获得三者的交集靶点。采用Cytoscape 3.9.1软件绘制“补肾启智方药物-活性成分-潜在靶点”网络、“补肾启智方-血管性痴呆-铁死亡靶点”的蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,并进行拓扑学分析以获得核心成分与核心靶点;在DAVID数据库进行交集靶点的基因本体(Gene Ontology,GO)功能富集分析、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析。采用AutoDock 4软件进行分子对接,并利用PyMol 2.5软件对分子对接结果进行可视化分析。通过动物实验验证关键靶点及信号通路。将32只SD雄性大鼠随机分为假手术组、模型组、补肾启智方高剂量组(14.4 g·kg^(-1))、补肾启智方低剂量组(3.6 g·kg^(-1)),每组8只。除假手术组外,其余大鼠采用双侧颈总动脉永久结扎(2-vessel occlusion,2-VO)法建立血管性痴呆模型。采用Morris水迷宫实验评估大鼠的空间学习与记忆能力;Western blot法检测大鼠海马组织中缺氧诱导因子1α(hypoxia inducible factor-1α,HIF-1α)、细胞溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)的蛋白表达;生化法检测大鼠海马组织中丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)水平。结果:筛选得到补肾启智方调控海马神经元铁死亡治�
Objective:To analyze and verify the mechanism of Bushen Qizhi Formula(BSQZF)in improving cognitive function of vascular dementia(VD)by regulating Ferroptosis in hippocampal neurons using network pharmacology and molecular docking technology combined with in vivo experiments.Methods:The active ingredients and their action targets of BSQZF were screened in TCMSP database,BATMAN-TCM database and ETCM database.Genecards database,OMIM database and DisGeNET database were used to screen potential targets of vascular dementia.FerrDB database was used to screen the Ferroptosis related targets,and all the above targets were standardized by Uniprot database.The intersection targets of the three were obtained from the website of Venn Diagrams.Cytoscape 3.9.1 software was used to draw the protein-protein interaction(PPI)network of"BSQZF drugs-active ingredients-potential targets"and"BSQZF-vascular dementia-Ferroptosis targets".Then the topological analysis was performed to obtain the core components and core targets.The DAVID database was used for Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.AutoDock4 software was used for molecular docking,and PyMol2.5 software was used to visualize the molecular docking results.The key targets and signaling pathways were verified by animal experiments.32 SD male rats were randomly divided into sham operation group,model group,BSQZF high-dose group(14.4 g·kg^(-1))and BSQZF low-dose group(3.6 g·kg^(-1)),with 8 rats in each group.Except the sham operation group,the vascular dementia model was established by permanent 2-vessel occlusion(2-VO).Morris water maze test was used to evaluate the spatial learning and memory ability of rats.Western blot was used to detect hypoxia inducible factor-1α(HIF-1α)and solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)in rat hippocampus.The levels of malondialdehyde(MDA)and glutathione(GSH)in hippocampus were detected by biochemical methods.Results:40 a
作者
巩俐
刘雪梅
刘云婷
王翎沣
刘孟涵
马承平
傅晨
GONG Li;LIU Xuemei;LIU Yunting;WANG Lingfeng;LIU Menghan;MA Chengping;FU Chen(Dongfang Hospital Affiliated to Beijing University of Chinese Medicine,Beijing China 100078)
出处
《中医学报》
CAS
2024年第8期1609-1621,共13页
Acta Chinese Medicine
基金
国家自然科学基金青年科学基金项目(82104808)。
