浙江中医药大学学报2025，Vol.49Issue(10)：1220-1232,13.DOI:10.16466/j.issn1005-5509.2025.10.002
基于网络药理学和体外实验探讨大黄抗脑缺血的作用机制
Exploring the Mechanism of Rhubarb root and rhizome in the Treatment of Ischemic Stroke by Network Pharmacology Analy-sis and in Vitro Experiment
摘要
Abstract
[Objective]To explore the anti-cerebral ischemia mechanism of Rhubarb root and rhizome by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)combined with network pharmacology and in vitro experiment.[Methods]The active components of Rhubarb root and rhizome in rat serum samples were identified by UPLC-Q-TOF/MS.The potential targets of Rhubarb root and rhizome in blood components were obtained by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Swiss Target Prediction.The related targets of ischemic stroke(IS)were obtained from Genome Annotation Database Platform(GeneCards),Online Mendelian Inheritance in Man(OMIM)database and Therapeutic Target Database(TTD).The intersection targets were obtained by using Venny 2.1.0 software and imported into the STRING database for protein-protein interaction(PPI)network analysis.Cytoscape 3.7.2 software was employed to visualize and identify the core targets.Gene ontology(GO)enrichment function analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signal pathway enrichment analysis were conducted to predict the mechanism of action.Molecular docking was performed by AutoDock Vina.Finally,the network pharmacological prediction results were verified by in vitro experiments.[Results]A total of 15 active components were identified in serum containing Rhubarb root and rhizome(SR),and there were 87 intersection targets between Rhubarb root and rhizome and IS.KEGG enrichment analysis showed that Rhubarb root and rhizome may alleviate IS through phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)and nuclear factor-κB(NF-κB)signaling pathways.Molecular docking results showed that the core components had a strong binding ability to the key targets of Toll-like receptor 4(TLR4)/myeloid differentiation primary response gene 88(MyD88)/NF-κB signaling pathway.In vitro experiment demonstrated that SR reduced the production of reactive oxygen species(ROS)induced by oxygen-glucose deprivation/reoxygenation(OGD/R)in BV2 cells.It also increased the activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),while decreasing the levels of malondialdehyde(MDA)and nitric oxide(NO).Additionally,SR reduced the levels of pro-inflammatory factors,increased the levels of anti-inflammatory factors in BV2 cells induced by OGD/R,and decreased the protein expression of TLR4,MyD88 and NF-κB in BV2 cells.[Conclusion]Rhubarb root and rhizome protects BV2 microglia from OGD/R-induced injury by inhibiting oxidative stress and inflammatory response,and its mechanism may be related to regulation of TLR4/MyD88/NF-κB signaling pathway.关键词
大黄/缺血性脑卒中/超高效液相色谱-四极杆-飞行时间质谱联用/网络药理学/BV2小胶质细胞/氧化应激/炎症反应/TLR4/MyD88/NF-κB信号通路Key words
Rhubarb root and rhizome/ischemic stroke/ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry/network pharmacology/BV2 microglial cells/oxidative stress/inflammatory response/TLR4/MyD88/NF-κB signaling pathway分类
医药卫生引用本文复制引用
曹兴琴,张诗晴,毛明江,张倩文,郭莹..基于网络药理学和体外实验探讨大黄抗脑缺血的作用机制[J].浙江中医药大学学报,2025,49(10):1220-1232,13.基金项目
浙江中医药大学2023年附属医院科研专项资助项目(2023FSYYZZ18) （2023FSYYZZ18）
浙江省中医药科研基金项目(2020ZB064) （2020ZB064）
浙江中医药大学生命科学学院科研培育项目(2022BJ002) （2022BJ002）
浙江省自然科学基金项目(LY16H280007) Research Project of Affiliated Hospital of Zhejiang Chinese Medical University(2023FSYYZZ18) （LY16H280007）
Zhejiang Traditional Chinese Medicine Science Research Foundation Project(2020ZB064) （2020ZB064）
Science Research Cultivation Project from College of Life Sciences of Zhejiang Chinese Medical University(2022BJ002) （2022BJ002）
Zhejiang Natural Science Foundation Project(LY16H280007) （LY16H280007）
