中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (36): 9519-9525.doi: 10.12307/2026.904

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

电针调控线粒体铜稳态干预缺血性脑卒中大鼠的神经炎性损伤

杨  娇1，王之枫2，施  婷2，徐双凤2，田大满2，蓝浚峯2，和鹏芬2，郝志慧2，潘  攀1，王  健2   

  1. 1云南中医药大学第二临床医学院，云南省昆明市  650500；2云南中医药大学第一临床医学院，云南省昆明市  650500
  • 收稿日期:2025-10-15 修回日期:2026-03-06 出版日期:2026-12-28 发布日期:2026-05-22
  • 通讯作者: 王健，博士，副教授，云南中医药大学第一临床医学院，云南省昆明市 650500 共同通讯作者：潘攀，博士，讲师，云南中医药大学第二临床医学院，云南省昆明市 650500
  • 作者简介:杨娇，女，2000年生，云南省丽江市人，汉族，云南中医药大学第二临床医学院在读硕士，主要从事脑卒中疾病的诊疗及临床研究。
  • 基金资助:
    云南省应用基础研究计划一般项目(202201AT070214)，项目负责人：王健；云南省科技厅-中医药应用基础研究联合专项资金项目(202001AZ070001-020，202301AZ070001-016)，项目负责人：王健；云南省两类人才项目(202205AD160024)，项目负责人：王健；云南省科学技术厅基础研究计划(202101AU070204，202301AZ070001-026)，项目负责人：潘攀；云南省针灸推拿防治脑病创新团队项目(ZTNB404)，项目负责人：王之枫；云南省针灸推拿防治脑病创新团队(ZTNB212)，项目负责人：杨娇；国家自然科学基金项目(82560932)，项目负责人：王健

Electroacupuncture regulates mitochondrial copper homeostasis to modulate neuroinflammatory injury in rats with ischemic stroke

Yang Jiao¹, Wang Zhifeng², Shi Ting², Xu Shuangfeng², Tian Daman², Lan Junfeng², He Pengfen², Hao Zhihui², Pan Pan¹, Wang Jian²   

  1. 1Second Clinical Medical College, Yunnan University of Chinese Medicine, Kunming 650500, Yunnan Province, China; 2First Clinical Medical College, Yunnan University of Chinese Medicine, Kunming 650500, Yunnan Province, China
  • Received:2025-10-15 Revised:2026-03-06 Online:2026-12-28 Published:2026-05-22
  • Contact: Wang Jian, PhD, Associate professor, First Clinical Medical College, Yunnan University of Chinese Medicine, Kunming 650500, Yunnan Province, China Co-corresponding author: Pan Pan, PhD, Lecturer, Second Clinical Medical College, Yunnan University of Chinese Medicine, Kunming 650500, Yunnan Province, China
  • About author:Yang Jiao, MS candidate, Second Clinical Medical College, Yunnan University of Chinese Medicine, Kunming 650500, Yunnan Province, China
  • Supported by:
    General Program of Yunnan Provincial Applied Basic Research Plan, No. 202201AT070214 (to WJ); Joint Special Fund for Applied Basic Research in Traditional Chinese Medicine, Yunnan Provincial Department of Science and Technology, Nos. 202001AZ070001-020 and 202301AZ070001-016 (both to WJ); Yunnan Provincial Two-Type Talent Project, No. 202205AD160024 (to WJ); Basic Research Program of Yunnan Provincial Department of Science and Technology, Nos. 202101AU070204 and 202301AZ070001-026 (both to PP); Yunnan Provincial Innovation Team for Acupuncture and Tuina in the Prevention and Treatment of Encephalopathy, Nos. ZTNB404 (to WZF) and ZTNB212 (to YJ); National Natural Science Foundation of China, No. 82560932 (to WJ) 

PDF

1

摘要：



文题释义：
缺血性脑卒中：一种常见的急性脑血管疾病，由于脑动脉阻塞导致局部脑组织血液供应中断，造成神经细胞缺血、缺氧性坏死。该病致死率与致残率极高，在幸存者中，高达75%的人面临后遗症问题。其核心病理机制包括兴奋性毒性、氧化应激、炎症反应及线粒体功能障碍等，这些过程相互交织，最终导致血脑屏障破坏、神经元死亡和神经功能缺损，为临床治疗带来巨大挑战。
铜稳态：指机体内铜元素的吸收、转运、分布、储存及排泄维持动态平衡的状态。铜是线粒体功能、抗氧化防御等关键生理过程的必需微量元素，参与能量代谢、抗氧化防御及信号转导。然而，在病理状态下铜稳态失衡会导致铜离子异常蓄积，通过芬顿反应产生活性氧自由基，诱发严重氧化应激与线粒体功能障碍，从而加剧细胞损伤，在神经退行性疾病及脑卒中过程中扮演重要角色。

背景：电针作为传统与现代结合的疗法，已被证实可改善脑卒中预后，但其能否通过调控线粒体铜稳态干预神经炎性损伤，机制尚不明确。
目的：观察电针刺激“百会”“大椎”对脑缺血再灌注损伤的治疗效应，并探讨其改善脑缺血再灌注损伤中铜稳态失衡的可能机制。
方法：将雄性SD大鼠随机分为假手术组、模型组和电针组，每组12只，后两组采用改良大脑中动脉闭塞法制备脑缺血再灌注损伤大鼠模型，电针组予针刺“百会”“大椎”，每日电针刺激1次，每次30 min，连续治疗14 d。Zea-Longa评分评估大鼠神经功能缺损程度；三苯基氯化四氮唑染色法测定脑组织梗死体积；苏木精-伊红染色和尼氏染色观察脑组织病理结构变化及红氨酸染色观察铜离子沉积情况；透射电镜观察大鼠皮质神经元超微结构；比色法检测梗死区脑组织中活性氧、铁硫簇以及亚铜离子水平；免疫组化染色和蛋白免疫印迹观察Kelch样ECH关联蛋白1、核因子红系2相关因子2、ATP酶铜转运蛋白β和二氢硫辛酰胺S-转乙酰基酶蛋白表达水平。
结果与结论：①与假手术组相比，模型组大鼠Zea-Longa评分升高(P < 0.001)，脑梗死体积百分比明显增加(P < 0.001)；梗死区神经元数量减少，排列松散，神经元呈明显病理变化，细胞器数量减少；脑组织中活性氧、亚铜离子含量显著升高(P < 0.001)，铁硫簇含量显著减少(P < 0.001)；皮质区核因子红系2相关因子2、ATP酶铜转运蛋白β和二氢硫辛酰胺S-转乙酰基酶蛋白表达显著降低(P < 0.05，P < 0.001，P < 0.01)，Kelch样ECH关联蛋白1蛋白表达显著上升(P < 0.001)；②与模型组相比，电针组大鼠Zea-Longa评分降低(P < 0.001)，脑梗死体积百分比降低(P < 0.05)；梗死区神经元数量增多，排列较规整，神经元核膜相对完整，线粒体、内质网等细胞器数量增多；脑组织中活性氧、亚铜离子含量显著降低(P < 0.05，P < 0.001)，铁硫簇含量升高(P < 0.01)；皮质中核因子红系2相关因子2、ATP酶铜转运蛋白β和二氢硫辛酰胺S-转乙酰基酶的蛋白表达显著升高(P < 0.05，P < 0.001，P < 0.05)，Kelch样ECH关联蛋白1蛋白表达显著下降(P < 0.05)；③提示电针可通过调控Kelch样ECH相关蛋白1-核因子红系2相关因子2信号通路，促进ATP酶铜转运蛋白β介导的线粒体铜离子转运，维持线粒体铜稳态与正常能量代谢，从而保护线粒体结构与功能完整性，最终改善缺血再灌注后的神经功能损伤，促进脑组织修复。
https://orcid.org/0009-0004-9206-0904(杨娇)


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 电针, 缺血性脑卒中, 铜稳态, Kelch样ECH关联蛋白1, Kelch样ECH相关蛋白1-核因子红系2相关因子2信号通路, 脑缺血再灌注损伤

Abstract: BACKGROUND: Electroacupuncture, a therapy that integrates traditional and modern elements, has been shown to improve the prognosis of stroke. However, whether it can intervene in neuroinflammatory injury by regulating mitochondrial copper homeostasis remains unclear.  
OBJECTIVE: To observe the therapeutic effects of electroacupuncture stimulation at “Baihui” and “Dazhui” on cerebral ischemia-reperfusion injury, and to explore the possible mechanisms by which it improves copper homeostasis imbalance in cerebral ischemia-reperfusion injury.  
METHODS: Male Sprague-Dawley rats were randomly divided into a sham-operated group, a model group, and an electroacupuncture group, with 12 rats in each group. The latter two groups were used to establish a rat model of cerebral ischemia-reperfusion injury using the modified middle cerebral artery occlusion method. The electroacupuncture group received acupuncture at “Baihui” and “Dazhui” acupoints, once daily for 30 minutes each time, for 14 consecutive days. Zea-Longa scores were used to assess the degree of neurological deficits. Triphenyltetrazolium chloride staining was used to measure cerebral infarct volume. Hematoxylin-eosin staining and Nissl staining were employed to observe pathological structural changes in brain tissue, and rhodanine staining was used to observe copper ion deposition. Transmission electron microscopy was used to observe the ultrastructure of rat cortical neurons. Colorimetric assays were used to detect the levels of reactive oxygen species, iron-sulfur clusters, and cuprous ions in the infarcted brain tissue. Immunohistochemical staining and western blot assay were used to observe the protein expression levels of Kelch-like ECH-associated protein 1 (Keap1), nuclear factor erythroid 2-related factor 2 (Nrf2), ATPase copper transporting beta (ATP7B), and dihydrolipoamide S-acetyltransferase (DLAT).  
RESULTS AND CONCLUSION: (1) Compared with the sham-operated group, the model group showed significantly increased Zea-Longa scores (P < 0.001) and a significantly increased percentage of cerebral infarct volume (P < 0.001); in the infarcted area, the number of neurons was reduced, their arrangement was loose, neurons showed obvious pathological changes, and the number of organelles was decreased; the contents of reactive oxygen species and cuprous ions in brain tissue were significantly increased (P < 0.001), while the content of iron-sulfur clusters was significantly decreased (P < 0.001); in the cortex, the protein expressions of Nrf2, ATP7B, and DLAT were significantly decreased (P < 0.05, P < 0.001, P < 0.01, respectively), and the protein expression of Keap1 was significantly increased (P < 0.001). (2) Compared with the model group, the electroacupuncture group showed significantly decreased Zea-Longa scores (P < 0.001) and a decreased percentage of cerebral infarct volume (P < 0.05); in the infarcted area, the number of neurons was increased, their arrangement was relatively regular, the nuclear membrane of neurons was relatively intact, and the number of organelles such as mitochondria and endoplasmic reticulum was increased; the contents of reactive oxygen species and cuprous ions in brain tissue were significantly decreased (P < 0.05, P < 0.001), and the content of iron-sulfur clusters was increased (P < 0.01); in the cortex, the protein expressions of Nrf2, ATP7B, and DLAT were significantly increased (P < 0.05, P < 0.001, P < 0.05, respectively), and the protein expression of Keap1 was significantly decreased (P < 0.05). To conclude, these findings suggest that electroacupuncture can protect mitochondrial structural and functional integrity by regulating the Keap1-Nrf2 signaling pathway, promoting ATP7B-mediated mitochondrial copper transport, and maintaining mitochondrial copper homeostasis and normal energy metabolism, ultimately improving neurological function impairment after ischemia-reperfusion and promoting brain tissue repair.  

Key words: electroacupuncture, ischemic stroke, copper homeostasis, Kelch-like ECH-associated protein 1, Kelch-like ECH-associated protein 1-nuclear factor erythroid 2-related factor 2 signaling pathway, cerebral ischemia-reperfusion injury

中图分类号: