中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (31): 8174-8180.doi: 10.12307/2026.488

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

电针干预脑缺血再灌注损伤诱导学习记忆障碍大鼠突触相关蛋白的变化

王慧灵1，2，郭长胜1，3，高  静1，黄  金4，申  昕1，赵  薇1，苏凯奇1，刘飞来1，冯晓东1，2   

  1. 1河南中医药大学第一附属医院康复诊疗中心，河南省郑州市   450000；2河南中医药大学第一临床医学院，河南省郑州市  450000 ；3黑龙江中医药大学，黑龙江省哈尔滨市   150040；4河南中医药大学康复医学院，河南省郑州市   450046
  • 收稿日期:2025-11-06 接受日期:2026-03-17 出版日期:2026-11-08 发布日期:2026-05-25
  • 通讯作者: 冯晓东，博士，主任医师，河南中医药大学第一附属医院康复诊疗中心，河南省郑州市 450000；河南中医药大学第一临床医学院， 河南省郑州市 450000
  • 作者简介:王慧灵，女，1989年生，河南省郑州市人，汉族，河南中医药大学在读博士，主治医师，主要从事常见功能障碍中医康复的临床与基础研究。
  • 基金资助:
    国家自然科学基金面上项目(82575189，82174473)，项目负责人：冯晓东；国家自然科学基金青年项目(82305364)，项目负责人：苏凯奇；河南省科技攻关项目(232102311210)，项目负责人：王慧灵；河南省科技攻关项目(222102310648)，项目负责人：申昕；中国博士后科学基金面上项目(2025M773962)，项目负责人：苏凯奇；河南省自然科学基金项目(262300421574)，项目负责人： 刘飞来

Alterations in synapse-associated proteins following electroacupuncture intervention in rats with learning and memory deficits induced by cerebral ischemia-reperfusion injury

Wang Huiling1, 2, Guo Changsheng1, 3, Gao Jing1, Huang Jin4, Shen Xin1, Zhao Wei1, Su Kaiqi1, Liu Feilai1, Feng Xiaodong1, 2   

  1. 1Rehabilitation Department of the First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000, Henan Province, China; 2The First Clinical Medical College of Henan University of Chinese Medicine, Zhengzhou 450000, Henan Province, China; 3Heilongjiang University of Chinese Medicine, Harbin 150040, Heilongjiang Province, China; 4College of Rehabilitation Medicine, Henan University of Chinese Medicine, Zhengzhou 450046, Henan Province, China
  • Received:2025-11-06 Accepted:2026-03-17 Online:2026-11-08 Published:2026-05-25
  • Contact: Feng Xiaodong, PhD, Chief physician, Rehabilitation Department of the First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000, Henan Province, China; The First Clinical Medical College of Henan University of Chinese Medicine, Zhengzhou 450000, Henan Province, China
  • About author:Wang Huiling, PhD candidate, Attending physician, Rehabilitation Department of the First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000, Henan Province, China; The First Clinical Medical College of Henan University of Chinese Medicine, Zhengzhou 450000, Henan Province, China
  • Supported by:
    National Natural Science Foundation of China (General Program), Nos. 82575189 and 82174473 (both to FXD); National Natural Science Foundation of China (Young Scientist Program), No. 82305364 (to SKQ); Science and Technology Research Project of Henan Province, No. 232102311210 (to WHL); Science and Technology Research Project of Henan Province, No. 222102310648 (to SX); China Postdoctoral Science Foundation (General Program), No. 2025M773962 (to SKQ); Natural Science Foundation of Henan Province, No. 262300421574 (to LFL)

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摘要：

文题释义：

脑缺血再灌注：是指脑组织在经历一定时间的缺血缺氧(血流中断或显著减少，导致氧气和营养供应不足)后，血流重新恢复(再灌注)的病理生理过程。
学习记忆障碍：指个体在获取(学习)、巩固(存储)或再现(提取)信息的过程中，出现持续性、显著性的功能减退或异常，表现为新信息学习能力下降、已存储记忆模糊或丢失、回忆准确性/速度降低，且这种障碍超出正常年龄相关衰退范围，并干扰日常生活或社会活动的病理性认知功能缺陷综合征。

摘要
背景：基础研究证实电针能改善大脑中动脉阻塞/再灌注模型大鼠的学习记忆能力，但其起效机制尚不清晰。
目的：观察电针“神庭”“百会”对大脑中动脉阻塞/再灌注后学习记忆障碍大鼠腺苷酸活化蛋白激酶(AMPK)/动力相关蛋白1(Drp1)通路相关蛋白及突触相关蛋白SYN，PSD95及GAP43表达的影响。
方法：采用随机化方法，将48只雄性SD大鼠分为空白组、假手术组、模型组和电针组，模型组和电针组大鼠以线栓法建立大脑中动脉阻塞/再灌注后学习记忆障碍大鼠模型。造模成功24 h后，电针干预电针组大鼠“神庭”“百会”，予疏密波，频率2 Hz/10 Hz，电流
1 mA，留针30 min，每日1次，共干预14 d。分别在造模后、干预第7，14天观察神经功能缺损评分；Morris水迷宫实验评估大鼠学习记忆能力；苏木精-伊红染色观察大鼠海马CA1区神经元形态；试剂盒测定三磷酸腺苷评估线粒体功能；透射电镜观察海马神经元中线粒体超微结构；Western blot检测海马组织AMPK、p-AMPK、Drp1、OPA1、Mfn1、Mfn2、SYN、PSD95、GAP-43蛋白表达。
结果与结论：①相较于空白组和假手术组，模型组和电针组大鼠的神经功能缺损评分均升高(P < 0.01)，干预7，14 d后，电针组大鼠神经功能缺损评分低于模型组(P < 0.05，P < 0.01)。②Morris水迷宫测试：相较于空白组及假手术组，模型组和电针组大鼠逃避潜伏期延长(P < 0.01)，穿越平台次数减少(P < 0. 01)；相对于模型组，电针组大鼠在电针干预后逃避潜伏期减少 (P < 0. 05)，穿越平台次数增加(P < 0. 01)。③苏木精-伊红染色显示，相对于模型组，电针组海马CA1区表现出神经元数目增多与排列更规则，结构趋于正常，细胞形态、大小较为均一；电镜观察可见，线粒体形态改善，结构破坏减轻，线粒体嵴数量增多。④ATP测试盒结果显示，与假手术组比较，模型组大鼠海马组织ATP含量降低，电针组大鼠海马组织ATP含量较模型组升高(P < 0.01)。⑤Western blot检测结果显示，模型组较假手术组海马的Drp1蛋白水平升高(P < 0.05)，p-AMPK/AMPK、OPA1、MFN1、MFN2、SYN、PSD95及GAP43蛋白水平下降(P < 0.01)；与模型组相比，电针组Drp1蛋白水平降低，而p-AMPK/AMPK、OPA1、MFN1、MFN2、SYN、PSD95和GAP43蛋白表达升高(P < 0.05)。⑥结果表明，电针能减轻大脑中动脉阻塞/再灌注大鼠的脑损伤，提高学习记忆能力，可能与腺苷酸活化蛋白激酶/动力相关蛋白1通路介导的线粒体动力学有关。  

关键词: 脑缺血再灌注损伤, 学习记忆障碍, 电针, 腺苷酸活化蛋白激酶/动力相关蛋白1, 线粒体动力学, 信号通路

Abstract: BACKGROUND: Basic research has confirmed that electroacupuncture can improve the learning and memory ability of rats with middle cerebral artery occlusion/reperfusion. However, its underlying mechanism remains unclear.  
OBJECTIVE: To observe the effects of electroacupuncture at “Shenting” and “Baihui” on the expression of adenosine monophosphate-activated protein kinase (AMPK)/dynamin-related protein 1 (Drp1) pathway-related proteins and synaptic proteins (SYN, PSD95, and GAP43) in rats with learning and memory impairment after middle cerebral artery occlusion/reperfusion.  
METHODS: A total of 48 male Sprague-Dawley rats were randomly assigned into four groups: control group, sham group, model group, and electroacupuncture group. The middle cerebral artery occlusion/reperfusion injury model was established by thread occlusion in the model group and the electroacupuncture group. At 24 hours after modeling, the rats in the electroacupuncture group were treated at “Shenting” and “Baihui” with disperse-dense wave, 2 Hz/10 Hz in frequency and 1 mA in current intensity, and the needles were retained for 30 minutes, once a day for 14 days. The neurological deficit scores were observed after modeling and on the 7th and 14th days of intervention. The learning and memory abilities of the rats were assessed using the Morris water maze test. Neuronal morphology in the rat hippocampal CA1 region was observed by hematoxylin-eosin staining. Adenosine triphosphate was measured by a kit to evaluate mitochondrial function. Transmission electron microscopy was used to examine the ultrastructure of neuronal mitochondria in the hippocampal region. The protein expressions of AMPK, p-AMPK, Drp1, OPA1, Mfn1, Mfn2, SYN, PSD95 and GAP-43 in the hippocampus were detected by western blot assay.  
RESULTS AND CONCLUSION: (1) Compared with the control group and sham group, the neurological deficit scores were increased in the model group and the electroacupuncture group (P < 0.01). After 7 and 14 days of intervention, the neurological deficit scores in the electroacupuncture group were lower than those in the model group (P < 0.05, P < 0.01). (2) The results of Morris water maze test: Compared with the control group and sham group, the model group and the electroacupuncture group showed increased escape latency (P < 0.01) and decreased platform crossings (P < 0.01). Furthermore, electroacupuncture treatment obviously decreased escape latency (P < 0.05) and increased platform crossings (P < 0.01) compared with the model groups. (3) Hematoxylin-eosin staining results showed that compared with the model group, the electroacupuncture group exhibited an increased number of neurons with a more regular arrangement in the hippocampal CA1 region. The structure appeared more normal, with relatively uniform neuronal cell size and morphology. Under the transmission electron microscopy, mitochondrial morphology was improved, characterized by reduced structural damage and an increased number of mitochondrial cristae. (4) Results from the adenosine triphosphate assay kit revealed a significant decrease in adenosine triphosphate content in the hippocampal tissue of the model group relative to the sham group. Meanwhile, the electroacupuncture group exhibited significantly elevated adenosine triphosphate levels compared with the model group (P < 0.01). (5) As revealed by western blot assay, the model group exhibited a significant increase in the protein level of Drp1 (P < 0.05), while showing significant decreases in the protein expressions of p-AMPK/AMPK, OPA1, MFN1, MFN2, SYN, PSD95 and GAP43 in the hippocampal tissue compared with the sham group (P < 0.01). In contrast, the electroacupuncture group displayed an opposite pattern compared with the model group, with decreased Drp1 level and increased expression of p-AMPK/AMPK, OPA1, MFN1, MFN2, SYN, PSD95 and GAP43 (P < 0.05). (6) To conclude, electroacupuncture treatment mitigates brain injury and enhances learning and memory ability in rats subjected to middle cerebral artery occlusion/reperfusion, likely through modulating mitochondrial dynamics mediated by the AMPK/Drp1 pathway.  

Key words: cerebral ischemia/reperfusion injury, learning and memory disorders, electroacupuncture, adenosine monophosphate-activated protein kinase/dynamic-related protein 1, mitochondrial dynamics, signaling pathway

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