北京中医药2025，Vol.44Issue(12)：1531-1542,12.DOI:10.16025/j.1674-1307.2025.12.006
基于网络药理学与代谢组学探讨补气摄血方对脑梗死溶栓后脑紧密连接的保护机制
Protective mechanisms of Buqi Shexue Formula on brain tight junctions after thrombolysis in cerebral infarction based on network pharmacology and metabolomics
摘要
Abstract
Objective To investigate the protective effects and mechanisms of Buqi Shexue Formula(BQSXF)on blood-brain barrier(BBB)tight junctions in rats with cerebral ischemia/reperfusion(CIR)injury after intravenous recombinant human tissue-type plasminogen activator(rt-PA),using combined untargeted metabolomics and network pharmacology.Methods A total of 105 rats were randomly divided into seven groups(n=15 each),including blank,sham-operated,model,atorvastatin,and BQSXF low-,medium-,and high-dose groups.Except for the blank and sham-operated groups,the other five groups were subjected to middle cerebral artery occlusion/reperfusion modeling using the intraluminal filament method and received intravenous rt-PA to simulate post-thrombolytic cerebral ischemia/reperfusion injury.Neurological deficit scores were assessed.Brain tissue structure was observed by hematoxylin-eosin staining.BBB ultrastructure was examined by transmission electron microscopy,and protein expression of Occludin,Claudin-5,and ZO-1 in brain tissue was measured by Western blot.Differential metabolites and metabolic pathways after BQSXF intervention were analyzed by untargeted metabolomics.Potential therapeutic targets were predicted by network pharmacology,and a"metabolite-reaction-enzyme-gene"network was constructed using the Metscape plugin.Results Compared with the sham-operated group,the model group showed significantly increased neurological deficit scores(P<0.05),disordered neuronal arrangement with shrunken nuclei and increased intercellular spacing,swollen endothelial cells,disrupted or absent tight junctions,and reduced Claudin-5,Occludin,and ZO-1 protein expression in the brain(P<0.01).Compared with the model group,the atorvastatin and BQSXF groups showed reduced neurological deficit scores(P<0.05),more regular neuronal arrangement with reduced intercellular spacing,improved endothelial swelling and tight junction integrity,and significantly increased Claudin-5,Occludin,and ZO-1 protein expression(P<0.01)in a dose-dependent manner.Metabolomics identified 39 differential metabolites,including arachidonic acid,eicosapentaenoic acid,docosapentaenoic acid,erucic acid,and oxidized glutathione,and 18 related metabolic pathways,including unsaturated fatty acid biosynthesis,ferroptosis,and pantothenic acid and coenzyme A biosynthesis.Network pharmacology predicted 187 potential therapeutic targets,including cellular tumor antigens,interleukin-1,interleukin-6,protein kinase B1,and tumor necrosis factor.The"metabolite-reaction-enzyme-gene"network identified arachidonic acid and eicosapentaenoic acid as core metabolites,with key targets including PLA2G4A,PTGS2,and 5-lipoxygenase;major pathways involved were arachidonic acid metabolism,prostaglandin formation from arachidonic acid,formation of putative anti-inflammatory metabolites from eicosapentaenoic acid,and leukotriene metabolism.Conclusion BQSXF may protect BBB integrity and improve outcomes in acute cerebral infarction by regulating arachidonic acid metabolism-related pathways via modulation of PTGS2 and 5-lipoxygenase,thereby reducing tight junction disruption.关键词
急性脑梗死/补气摄血方/紧密连接/代谢组学/网络药理学/大鼠Key words
acute cerebral infarction/Buqi Shexue Formula/tight junction/metabolomics/network pharmacology/rats引用本文复制引用
赵聪,王革生,马涛,李文京,刘乙兴,袁健..基于网络药理学与代谢组学探讨补气摄血方对脑梗死溶栓后脑紧密连接的保护机制[J].北京中医药,2025,44(12):1531-1542,12.基金项目
国家自然科学基金面上项目(82374275) （82374275）
国家重点研发计划项目(2022YFC3501102) （2022YFC3501102）
