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- 北京中医药 2025年44卷第12期 页码：1531-1542
- 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：
赵聪，男，29岁，博士。研究方向：中西医结合防治脑血管病。
王革生，E-mail：wanggesheng@bucm.edu.cn
- 基金信息：国家自然科学基金面上项目(82374275);国家重点研发计划项目(2022YFC3501102)
DOI：10.16025/j.1674-1307.2025.12.006
中图分类号：收稿：2024-12-05，
纸质出版：2025-12-25
- 稿件说明：
移动端阅览
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
2
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
2
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
2
与假手术组比较，模型组神经功能缺损评分明显升高（
P
<
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（
P
<
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（
P
<
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（
P
<
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
2
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
2
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
2
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
2
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 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
2
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
中华医学会神经病学分会 , 中华医学会神经病学分会脑血管病学组 . 中国急性缺血性卒中诊治指南2023 [J ] . 中华神经科杂志 ， 2024 , 57 ( 6 )： 523 - 559 .
ROTH JM . Recombinant tissue plasminogen activator for the treatment of acute ischemic stroke [J ] . Proc (Bayl Univ Med Cent) , 2011 , 24 ( 3 ): 257 - 259 .
KHEDR EM , ABDELWARITH A , MOUSSA G , et al . Recombinant tissue plasminogen activator (rTPA) management for first onset acute ischemic stroke with covid -19 and non-covid -19 patients [J ] . J Stroke Cerebrovasc Dis , 2023 , 32 ( 4 ): 107031 .
ZHANG S , WANG D , LI L . Recombinant tissue-type plasminogen activator (rt-PA) effectively restores neurological function and improves prognosis in acute ischemic stroke [J ] . Am J Transl Res , 2023 , 15 ( 5 ): 3460 - 3467 .
王维 , 房砚文 , 宫萍 . 急性轻型缺血性卒中患者静脉溶栓后早期神经功能恶化的危险因素分析 [J ] . 解放军医学杂志 , 2024 , 49 ( 6 ): 617 - 622 .
陈燕 , 张秋 , 尤年兴 , 等 . 急性缺血性脑卒中静脉溶栓后神经功能恢复及脑出血转化发生的影响因素分析 [J ] . 中华危重病急救医学 , 2020 , 32 ( 11 ): 1340 - 1345 .
孔令雷 , 马寅仲 , 李莉 , 陈燕霞 , 杜冠华 . 脑卒中溶栓后出血转化的发生机制及治疗药物研究进展 . 药学学报 , 2018 , 53 ( 9 ): 1467 - 1476 .
LOCHHEAD JJ , YANG J , RONALDSON PT , et al . Structure, Function, and Regulation of the Blood-Brain Barrier Tight Junction in Central Nervous System Disorders [J ] . Front Physiol , 2020 , 11 : 914 .
ZHENG X , REN B , GAO Y . Tight junction proteins related to blood-brain barrier and their regulatory signaling pathways in ischemic stroke [J ] . Biomed Pharmacother , 2023 , 165 : 115272 .
中国中西医结合学会 , 中华中医药学会 , 中华医学会 . 脑梗死中西医结合诊疗指南 [J ] . 中国中医药信息杂志 , 2023 , 40 ( 5 ): 1 - 82 .
陈可冀 . 血瘀证与活血化瘀治疗的研究 [J ] . 中国中医药现代远程教育 , 2005 , 3 ( 11 ): 10 - 12 .
付于 , 黄燕 , 黄培新 , 等 . 急性缺血性脑血管病的中药溶栓治疗与代谢组学的研究趋势 [J ] . 辽宁中医杂志 , 2006 , 33 ( 6 ): 662 - 664 .
吴婧 , 张志辰 , 李中浩 , 等 . 急性脑梗死患者静脉溶栓后中医证候要素演变 [J ] . 北京中医药大学学报 , 2020 , 43 ( 1 ): 74 - 78 .
曹华政 , 马登峰 , 李双成 , 等 . 从气虚血瘀理论研究益气活血法防治急性缺血性脑卒中的出血转化机制 [J ] . 四川中医 , 2023 , 41 ( 10 ): 26 - 29 .
黄世敬 . 基于神经血管单元失稳态探讨气虚留滞病机 [J ] . 世界中西医结合杂志 , 2016 , 11 ( 6 ): 855 - 858，862 .
刘深 , 张兆旭 , 高颖 . 补气摄血法防治脑梗死溶栓后出血转化探析 [J ] . 北京中医药 , 2022 , 41 ( 7 ): 725 - 727 .
张翔 , 刘从国 , 张继红 , 等 . 益气活血中药联合西医治疗急性缺血性脑卒中的Meta分析 [J ] . 中国老年学杂志 , 2017 , 37 ( 4 ): 873 - 876 .
徐央丽 . 中药煎煮基本原则及操作规程 [J ] . 浙江中医药大学学报 , 2007 , 31 ( 5 ): 624 - 625 .
LONGA EZ , WEINSTEIN PR , CARLSON S , et al . Reversible middle cerebral artery occlusion without craniectomy in rats [J ] . Stroke , 1989 , 20 ( 1 ): 84 - 91 .
ULUç K , MIRANPURI A , KUJOTH GC , et al . Focal cerebral ischemia model by endovascular suture occlusion of the middle cerebral artery in the rat [J ] . J Vis Exp , 2011 ( 48 ): 1978 .
陈奇 ， 孙建宁 ， 林志彬 ， 等 . 中药药理研究方法学 [M ] . 3版 . 北京 ： 人民卫生出版社 ， 2011 : 45 – 47 .
LU M , CHEN J , LU D , et al . Global test statistics for treatment effect of stroke and traumatic brain injury in rats with administration of bone marrow stromal cells [J ] . J Neurosci Methods , 2003 , 128 ( 1-2 ): 183 - 190 .
周玉英 , 顾勇 . 组织型纤溶酶原激活剂溶栓后出血转化的机制及中医药防治的研究进展 [J ] . 中西医结合心脑血管病杂志 , 2023 , 21 ( 24 ): 4549 - 4554 .
李菁晶 , 梁德君 , 陈立云 , 等 . rt-PA联合亚低温治疗对缺血性脑卒中出血转化的干预研究 [J ] . 中国康复理论与实践 , 2008 , 14 ( 6 ): 521 - 523 .
张玉鹏 , 李芳 , 陈竹 , 等 . PJ34对脑缺血再灌注大鼠应用rt-PA后血脑屏障完整性的影响 [J ] . 中国临床解剖学杂志 , 2019 , 37 ( 3 ): 287 - 291,298 .
刘明亮 , 杨森林 , 马超 . 阿替普酶联合红花提取物调节TLR4-NLRP3信号通路对大鼠急性脑梗塞的影响 [J ] . 河北医学 , 2023 , 29 ( 9 ): 1426 - 1432 .
冯晓燕 , 李蓉 , 马涛 . 低剂量坎地沙坦对小鼠脑梗死使用重组组织型纤溶酶原激活剂溶栓的出血并发症的影响及机制 [J ] . 临床神经病学杂志 , 2020 , 33 ( 1 ): 52 - 56 .
贺涓涓 , 洪华 , 杨世亮 . 阿托伐他汀对脑缺血再灌注大鼠血脑屏障的保护作用 [J ] . 中国病理生理杂志 , 2015 , 31 ( 2 ): 219 - 223 .
DJURICIC I , CALDER PC . Beneficial outcomes of Omega-6 and Omega-3 polyunsaturated fatty acids on human health: an update for 2021 [J ] . Nutrients , 2021 , 13 ( 7 ).
BAZINET RP , LAYÉ S . Polyunsaturated fatty acids and their metabolites in brain function and disease [J ] . Nat Rev Neurosci , 2014 , 15 ( 12 ): 771 - 785 .
KAGAN VE , MAO G , QU F , et al . Oxidized arachidonic and adrenic PEs navigate cells to ferroptosis [J ] . Nat Chem Biol , 2017 , 13 ( 1 ): 81 - 90 .
SERHAN CN , LEVY BD . Resolvins in inflammation: emergence of the pro-resolving superfamily of mediators [J ] . J Clin Invest , 2018 , 128 ( 7 ): 2657 - 2669 .
HANNA VS , HAFEZ E . Synopsis of arachidonic acid metabolism: A review [J ] . J Adv Res , 2018 , 11 : 23 - 32 .
蒋欢 , 白文娅 , 邵建林 . 铁死亡在脑缺血再灌注损伤机制中的研究进展 [J ] . 中国比较医学杂志 , 2024 , 34 ( 7 ): 101 - 109 .
李光硕 , 赵性泉 . 《中国急性缺血性卒中诊治指南2023》解读 [J ] . 中国卒中杂志 , 2024 , 19 ( 8 ): 956 - 961 .
都一鸣 , 陈鑫 , 赵世光 . 急性缺血性脑卒中氧化应激机制的研究进展 [J ] . 中华神经创伤外科电子杂志 , 2021 , 7 ( 2 ): 121 - 124 .
BARRITT SA , DUBOIS-COYNE SE , DIBBLE CC . Coenzyme A biosynthesis: mechanisms of regulation, function and disease [J ] . Nat Metab , 2024 , 6 ( 6 ): 1008 - 1023 .
LI Y , LIAO J , XIONG L , et al . Stepwise targeted strategies for improving neurological function by inhibiting oxidative stress levels and inflammation following ischemic stroke [J ] . J 空白 Release , 2024 , 368 : 607 - 622 .
WANG B , WU L , CHEN J , et al . Metabolism pathways of arachidonic acids: mechanisms and potential therapeutic targets [J ] . Signal Transduct Target Ther , 2021 , 6 ( 1 ): 94 .
SARKAR C , JONES JW , HEGDEKAR N , et al . PLA2G4A/cPLA2-mediated lysosomal membrane damage leads to inhibition of autophagy and neurodegeneration after brain trauma [J ] . Autophagy , 2020 , 16 ( 3 ): 466 - 485 .
CHU LY , LIOU JY , WU KK . Prostacyclin protects vascular integrity via PPAR/14-3-3 pathway [J ] . Prostaglandins Other Lipid Mediat , 2015 , 118-119 : 19 - 27 .
LI L , SLUTER MN , YU Y , et al . Prostaglandin E receptors as targets for ischemic stroke: novel evidence and molecular mechanisms of efficacy [J ] . Pharmacol Res , 2021 , 163 : 105238 .
LEI B , WU H , YOU G , et al . Silencing of ALOX15 reduces ferroptosis and inflammation induced by cerebral ischemia-reperfusion by regulating PHD2/HIF2α signaling pathway [J ] . Biotechnol Genet Eng Rev , 2024 , 40 ( 4 ): 4341 - 4360 .
BORSINI A , NICOLAOU A , CAMACHO-MUñOZ D , et al . Omega-3 polyunsaturated fatty acids protect against inflammation through production of LOX and CYP450 lipid mediators: relevance for major depression and for human hippocampal neurogenesis [J ] . Mol Psychiatry , 2021 , 26 ( 11 ): 6773 - 6788 .
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