中医学报2024，Vol.39Issue(8)：1609-1621,13.DOI:10.16368/j.issn.1674-8999.2024.08.265
基于网络药理学和动物实验探讨补肾启智方对血管性痴呆铁死亡的作用机制
Mechanism of Bushen Qizhi Formula on Ferroptosis in Vascular Dementia Based on Network Pharmacology and Animal Experiments
摘要
Abstract
Objective:To analyze and verify the mechanism of Bushen Qizhi Formula(BSQZF)in improving cognitive function of vascu-lar 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 data-base,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 ratswererandomlydividedintoshamoperationgroup,modelgroup,BSQZFhigh-dosegroup(14.4 g·kg-1)and BSQZFlow-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 per-manent2-vesselocclusion(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 glu-tathione peroxidase 4(GPX4)in rat hippocampus.The levels of malondialdehyde(MDA)and glutathione(GSH)in hippocampus were detected by biochemical methods.Results:40 active ingredients and 30 effective targets of BSQZF for regulating Ferroptosis of hip-pocampal neurons in the treatment of VD were screened out.The core ingredients were quercetin,jasmonone and aristolochia,and the core targets were tumor protein P53(TP53),nuclear factor erythroid 2-related factor 2(Nrf2),mammalian target of rapamycin(MTOR),HIF-1α.According to the results of GO enrichment of function analysis,BSQZF could treat VD by improving the response of cells to hydrogen peroxide,hypoxia,oxidative stress and promoting angiogenesis.KEGG pathway enrichment analysis suggests that BS-QZF may regulate hypoxia inducing factor1(HIF-1)signaling pathway,cell senescence,reactive oxygen species,forkhead box protein O(FoxO)signaling pathway,epidermal growth factor receptor(EGFR)signaling pathway,etc.to treat VD.After treatment with Bushen Qizhi Decoction,the spatial learning and memory ability of VD rats was improved.The expression of MDA in hippocampus was de-creased significantly(P<0.01),while GSH level was increased significantly(P<0.01).Besides,the expression levels of HIF-1α and ferroptotic core regulatory proteins SLC7A11 as well as GPX4 were significantly increased(P<0.01,P<0.05,P<0.05)Conclu-sion:Bushen Qizhi Formula can improve the cognitive function of VD through multi-components and multi-targets,and its key mech-anism may be related to activation of HIF-1α/SLC7A11/GPX4 signaling pathway and inhibition of Ferroptosis in hippocampal neu-rons.关键词
补肾启智方/血管性痴呆/铁死亡/网络药理学/分子对接/HIF-1α/SLC7A11/GPX4信号通路/大鼠Key words
Bushen Qizhi Formula/vascular dementia/ferroptosis/network pharmacology/molecular docking/HIF1α/SLC7A11/GPX4 signaling pathway/rat分类
医药卫生引用本文复制引用
巩俐,刘雪梅,刘云婷,王翎沣,刘孟涵,马承平,傅晨..基于网络药理学和动物实验探讨补肾启智方对血管性痴呆铁死亡的作用机制[J].中医学报,2024,39(8):1609-1621,13.基金项目
国家自然科学基金青年科学基金项目(82104808) （82104808）
