文献类型：期刊文献

中文题名：基于网络药理学和实验验证探讨藏药二十味沉香丸治疗缺血性脑梗死的机制

英文题名：Exploring the Mechanism of Tibetan Medicine Ershiwai Chenxiang Wan in Treating Ischemic Stroke Based on Network Pharmacology and Experimental Verification

作者：卓玛草[1];贡保东知[2];曹旺杰[3];公保甲[2];加羊加措[2];尼玛次仁[1]

第一作者：卓玛草

机构：[1]西藏藏医药大学研究生处,拉萨850000;[2]甘肃中医药大学藏医学院,甘南747000;[3]甘肃中医药大学基础医学院,兰州730000

第一机构：西藏藏医药大学研究生处,拉萨850000

年份：2026

卷号：28

期号：5

起止页码：1546

中文期刊名：世界科学技术-中医药现代化

外文期刊名：Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

收录：;北大核心:【北大核心2023】；

基金：2023年度中医(藏医)博士点建设及中药学(藏药)博士点培育项目之博士研究生科研创新能力提高项目(BSDJS-BS-2023002),负责人:米玛;甘肃省教育厅青年博士基金(2025QB-071):基于数据挖掘技术整理和探究藏药止咳化痰方剂用药规律及剂型创新研究,负责人:贡保东知;甘肃(甘南)藏医药“医-药-护”研究与转化协同创新中心研究成果,负责人:黎昱民。

语种：中文

中文关键词：缺血性脑梗死;二十味沉香丸;网络药理;实验验证

外文关键词：Ischemic stroke;Ershiwei Chenxiang Wan;Network pharmacology;Experimental verification

摘要：目的研究藏药二十味沉香丸(Twenty-ingredient eaglewood pills,ECW)治疗缺血性脑梗死(Ischemic cerebral infarction)的机制。方法首先,基于前期超高效液相色谱法(Ultra-high performance liquid chromatography,UPLC)-四极杆(Quadrupole rod,Q)-飞行时间质谱仪(Time-of-flight mass spectrometer,TOFMS)鉴定的二十味沉香丸化学成分,利用SwissADME和TCMSP筛选具有良好类药性的活性成分,并通过SwissTargetPrediction预测其潜在作用靶点。从GeneCards数据库获取AICI相关靶点,取交集靶点作为二十味沉香丸治疗缺血性脑梗死的潜在靶点。利用STRING数据库构建蛋白质-蛋白质相互作用(Proteinprotein interaction,PPI)网络,筛选核心靶点。通过DAVID数据库对交集靶点进行基因本体(Gene ontology,GO)功能注释与京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析。采用分子对接技术对所选核心靶点和化合物进行分子对接验证。体外实验利用小鼠小胶质细胞(BV2)建立氧糖剥夺/复氧(Oxygen glucose deprivation/reoxygenation,OGD/R)损伤模型,通过CCK-8法检测细胞活力,流式细胞术检测细胞凋亡和活性氧(Reactive oxygen species,ROS)水平,蛋白质印迹法(Western blot)检测关键蛋白的表达变化,评估不同浓度含药血清对OGD/R损伤的保护作用。结果网络药理学分析筛选出27个活性成分和120个药物与疾病共同靶点。PPI网络及拓扑分析确定信号转导及转录激活蛋白3(Signal transducer and activator of transcription 3,STAT3)、促分裂原活化的蛋白激酶(Mitogenactivated protein kinase 3,MAPK3)、AKT1、表皮生长因子受体(Epidermal growth factor receptor,EGFR)、JAK、Cleaved-caspase3等为核心靶点。GO和KEGG富集分析表明这些靶点显著富集于炎症反应、凋亡过程负调控、谷氨酸能突触及帕金森病等通路。分子对接显示槲皮素、齐墩果酸、沉香四醇、愈创木酚基二氢松柏醇醚、四氢姜黄素等活性成分与JAK、STAT3、EGFR、Cleaved-caspase3等核心靶点具有较好的结合能。体外实验证实,与OGD/R模型组相比,二十味沉香丸含药血清能剂量依赖性地:①显著提高BV2细胞活力(CCK-8法);②显著降低OGD/R诱导的细胞凋亡率(流式细胞术);③显著减少细胞内ROS水平(流式细胞术);④显著下调p-JAK、p-STAT3、EGFR及Cleaved-caspase3(35 kDa和15 kDa)的蛋白表达(Western blot)。结论本研究提示,二十味沉香丸可能通过抑制JAK2/STAT3、EGFR信号通路活化及下游Caspase-3凋亡蛋白的激活,有效减轻OGD/R诱导的BV2小胶质细胞氧化应激和凋亡,从而在细胞水平发挥神经保护作用。
Objective To investigate the mechanism of the Tibetan medicine Ershiwai Chenxiang Wan(ECW Pills)in treating Acute ischemic cerebral infarction ischemic cerebral infarction(AICI).Methods Firstly,based on the chemical components of ECW previously identified by UPLC-Q-TOF/MS,active ingredients with favorable drug-likeness were screened using SwissADME and TCMSP.Their potential targets were predicted via SwissTargetPrediction.Diseaserelated targets for AICI were retrieved from the GeneCards database.Intersection targets between drug and disease targets were identified as potential therapeutic targets for ECW against AICI.A protein-protein interaction(PPI)network was constructed using the STRING database to screen core targets.Gene Ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis of the intersection targets were performed using the DAVID database.Molecular docking was employed to validate the binding between selected core targets and compounds.In vitro,an oxygen-glucose deprivation/reoxygenation(OGD/R)injury model was established using murine microglial cells(BV2).Cell viability was assessed by CCK-8 assay;apoptosis and reactive oxygen species(ROS)levels were detected by flow cytometry;expression changes of key proteins were measured by Western blotting.The protective effects of different concentrations of ECW-medicated serum against OGD/R injury were evaluated.Results Network pharmacology analysis screened 27 active components and 120 common drug-disease targets.PPI network and topological analysis identified STAT3,MAPK3,AKT1,EGFR,JAK,and Cleaved-caspase 3 as core targets.GO and KEGG enrichment analyses indicated that these targets were significantly enriched in pathways such as inflammatory response,negative regulation of apoptotic process,glutamatergic synapse,and Parkinson’s disease.Molecular docking demonstrated favorable binding affinities between active components(quercetin,oleanolic acid,agarotetrol,guaiacylglycerol-β-coniferyl ether and tetrahydrocurcumin)and core targets(JAK,STAT3,EGFR,Cleaved-caspase 3).In vitro experiments confirmed that,compared to the OGD/R model group,ECW-medicated serum dose-dependently:①Significantly increased BV2 cell viability(CCK-8 assay);②Significantly reduced the OGD/R-induced apoptosis rate(flow cytometry);③Significantly decreased intracellular ROS levels(flow cytometry);④Significantly downregulated the protein expression of phospho-JAK(p-JAK),phospho-STAT3(p-STAT3),EGFR and Cleavedcaspase 3(35 kDa and 15 kDa fragments)(Western blotting).Conclusion This study suggests that Ershiwai Chenxiang Wan may exert neuroprotective effects at the cellular level by mitigating OGD/R-induced oxidative stress and apoptosis in BV2 microglial cells.This protection is likely mediated through the inhibition of JAK2/STAT3 and EGFR signaling pathway activation and the subsequent suppression of downstream Caspase-3 apoptotic protein cleavage.