Protective mechanisms of <italic style="font-style: italic">Buqi Shexue Formula</italic> on brain tight junctions after thrombolysis in cerebral infarction based on network pharmacology and metabolomics

ZHAO Cong; WANG Gesheng; MA Tao; LI Wenjing; LIU Yixing; YUAN Jian

doi:10.16025/j.1674-1307.2025.12.006

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Protective mechanisms of Buqi Shexue Formula on brain tight junctions after thrombolysis in cerebral infarction based on network pharmacology and metabolomics

更新时间：2026-01-24

- Protective mechanisms of Buqi Shexue Formula on brain tight junctions after thrombolysis in cerebral infarction based on network pharmacology and metabolomics
- Beijing Journal of Traditional Chinese Medicine Vol. 44, Issue 12, Pages: 1531-1542(2025)
- 作者机构：
  
  1.北京中医药大学第二临床医学院，北京 100078
  2.北京中医药大学东方医院外三科，北京 100078
  3.北京中医药大学东方医院科研处，北京 100078
- 作者简介：
- 基金信息：
- DOI：10.16025/j.1674-1307.2025.12.006
  CLC：
- Received：05 December 2024，
  
  Published：25 December 2025
- 稿件说明：
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赵聪，王革生，马涛，等.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.

ZHAO Cong,WANG Gesheng,MA Tao,et al.Protective mechanisms of Buqi Shexue Formula on brain tight junctions after thrombolysis in cerebral infarction based on network pharmacology and metabolomics[J]. Beijing Journal of Traditional Chinese Medicine,2025,44(12):1531-1542.
赵聪，王革生，马涛，等.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. DOI： 10.16025/j.1674-1307.2025.12.006.

ZHAO Cong,WANG Gesheng,MA Tao,et al.Protective mechanisms of Buqi Shexue Formula on brain tight junctions after thrombolysis in cerebral infarction based on network pharmacology and metabolomics[J]. Beijing Journal of Traditional Chinese Medicine,2025,44(12):1531-1542. DOI： 10.16025/j.1674-1307.2025.12.006.

摘要

目的

基于非靶向代谢组学和网络药理学探讨补气摄血方对脑缺血再灌注大鼠应用重组人组织型纤溶酶原激活物（rt-PA）后的血脑屏障紧密连接的保护作用及机制。

方法

将105只SD大鼠随机分为空白组、假手术组、模型组、阿托伐他汀组、补气摄血方低剂量组、补气摄血方中剂量组、补气摄血方高剂量组，每组15只。除空白组及假手术组外，其余5组均采用线栓法建立大脑中动脉栓塞再灌注模型并静脉注射rt-PA，模拟临床脑梗死溶栓治疗后的脑缺血-再灌注损伤过程。观察并记录神经功能缺损评分，苏木精-伊红染色观察脑组织结构变化，透射电镜观察血脑屏障超微结构变化，Western blotting法检测脑组织大鼠脑组织闭合蛋白（Occludin）、密封蛋白-5（Claudin-5）、闭锁连接蛋白-1（ZO-1）蛋白表达。通过非靶向代谢组学分析补气摄血方干预后差异代谢产物及代谢通路，通过网络药理学预测补气摄血方干预急性脑梗死的潜在靶点，并通过Metscape插件构建“代谢物-反应-酶-基因”关系网络。

结果

与假手术组比较，模型组神经功能缺损评分明显升高（

0.05）；大鼠脑组织神经元细胞排列不规则，细胞核固缩且分离度增加；内皮细胞肿胀，紧密连接结构出现断裂和消失；脑组织Claudin-5、Occludin、ZO-1蛋白表达水平降低（

0.01）。与模型组比较，阿托伐他汀组、补气摄血方组大鼠神经功能缺损评分降低（

0.05）；神经元细胞排列较整齐，分离度减小；内皮细胞肿胀及紧密连接结构断裂、消失情况好转；脑组织Claudin-5、Occludin、ZO-1蛋白表达升高（

0.01），且升高幅度呈剂量依赖性。代谢组学筛选出花生四烯酸、二十碳五烯酸、二十二碳五烯酸、芥酸、氧化谷胱甘肽等39个差异代谢物和不饱和脂肪酸的生物合成、铁死亡、泛酸和辅酶A生物合成等18条代谢通路。网络药理学预测潜在治疗靶点为细胞肿瘤抗原、白细胞介素-1、白细胞介素-6、蛋白激酶B1、肿瘤坏死因子等共187个。“代谢物-反应-酶-基因”关系网络显示：核心代谢物为花生四烯酸和二十碳五烯酸。关键靶点包括磷脂酶A2组IVA、前列腺素内过氧化物合酶2及5-脂氧合酶；主要通路涉及花生四烯酸代谢、花生四烯酸衍生前列腺素、二十碳五烯酸来源的抗炎代谢产物形成以及白三烯代谢等。

结论

补气摄血方可能通过干预前列腺素内过氧化物合酶2、5-脂氧合酶等调节花生四烯酸代谢相关途径以减少紧密连接破坏，保护血脑屏障稳定性，进而改善急性脑梗死大鼠的预后。

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 （

=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 （

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 （

0.01）. Compared with the model group， the atorvastatin and BQSXF groups showed reduced neurological deficit scores （

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 （

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 eicosape

ntaenoic 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

Keywords

references

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⁰

Protective mechanisms of Buqi Shexue Formula on brain tight junctions after thrombolysis in cerebral infarction based on network pharmacology and metabolomics

DOI：10.16025/j.1674-1307.2025.12.006

摘要

Abstract

关键词

Keywords

references