中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (14): 2186-2190.doi: 10.12307/2024.278

• 骨组织构建 bone tissue construction • 上一篇    下一篇

电针对脑缺血再灌注损伤大鼠Toll样受体4/核因子κB信号通路的影响

罗  敷1，2，3，舒象忠4，刘丹妮1，2，3，谭金曲1，2，3，彭  婷1，2，3，黄夏荣1，2，3，孙光华1，2，3，彭昕珂1，2，3，王金玲1，2，3，周  君1，2，3   

  1. 南华大学衡阳医学院附属第一医院，1康复医学科，2康复医学中心，3康复医学实验室，4放射医学科，湖南省衡阳市  421001
  • 收稿日期:2023-02-02 接受日期:2023-03-22 出版日期:2024-05-18 发布日期:2023-07-28
  • 通讯作者: 周君，博士后，主任医师，南华大学衡阳医学院附属第一医院，康复医学科，康复医学中心，康复医学实验室，湖南省衡阳市 421001
  • 作者简介:罗敷，女，1991年生，汉族，2017年湖南中医药大学毕业，硕士，主治医师，主要从事神经系统疾病中医传统康复治疗基础及临床研究。
  • 基金资助:
    湖南省自然科学基金(2023JJ40590)，项目负责人：罗敷；国家自然科学基金(81973917)，项目负责人：周君；湖南省研究生科研创新项目(CX20210973)，项目负责人：刘丹妮

Electroacupuncture reduces inflammatory factor expression by suppressing Toll-like receptor 4/nuclear factor-kappa B signaling in rats with cerebral ischemia-reperfusion injury

Luo Fu1, 2, 3, Shu Xiangzhong4, Liu Danni1, 2, 3, Tan Jinqu1, 2, 3, Peng Ting1, 2, 3, Huang Xiarong1, 2, 3, Sun Guanghua1, 2, 3, Peng Xinke1, 2, 3, Wang Jinling1, 2, 3, Zhou Jun1, 2, 3   

  1. 1Department of Rehabilitation, 2Rehabilitation Medicine Center, 3Rehabilitation Laboratory, 4Department of Radiological Medicine, The First Affiliated Hospital of Hengyang Medical School, University of South China, Hengyang 421001, Hunan Province, China
  • Received:2023-02-02 Accepted:2023-03-22 Online:2024-05-18 Published:2023-07-28
  • Contact: Zhou Jun, MD, Chief physician, Department of Rehabilitation, Rehabilitation Medicine Center, Rehabilitation Laboratory, The First Affiliated Hospital of Hengyang Medical School, University of South China, Hengyang 421001, Hunan Province, China
  • About author:Luo Fu, Master, Attending physician, Department of Rehabilitation, Rehabilitation Medicine Center, Rehabilitation Laboratory, The First Affiliated Hospital of Hengyang Medical School, University of South China, Hengyang 421001, Hunan Province, China
  • Supported by:
    Hunan Province Natural Science Foundation, No. 2023JJ40590 (to LF); National Natural Science Foundation of China, No. 81973917 (to ZJ); Hunan Provincial Postgraduate Research Innovation Project, No. CX20210973 (to LDN)

PDF

34

摘要：


文题释义：

脑缺血再灌注损伤：是指在缺血脑组织供血恢复后发生的继发性脑损伤。它的机制可能包括自由基损伤、神经炎症反应、胞内钙离子超载、血脑屏障破坏、神经细胞程序性死亡等。神经炎症反应是脑缺血再灌注损伤时经常发生的事件，表现为脑内的星形胶质细胞和小胶质细胞过度激活、炎性细胞因子和趋化因子产生过多、血液中的中性粒细胞等浸润到受损脑组织内的一系列事件。最终，过度的炎症级联反应导致神经元死亡并加重脑损伤。
Toll样受体4/核因子κB信号通路：Toll样受体是一种可介导免疫炎症反应发生的模式识别受体。Toll样受体4是一种Toll同源体，可介导核转录因子κB激活，诱导促炎性因子的大量产生，启动炎性应答。核转录因子κB参与众多免疫细胞应答、炎症因子激活及细胞退变与凋亡，对于启动基因的转录及与靶点位的特异性结合发挥着重要作用，与炎症反应程度呈正相关。


背景：炎症反应是诱发脑缺血再灌注损伤的重要因素之一。研究表明电针可有效降低缺血性脑卒中后炎症反应，改善神经功能缺损症状，但机制尚未明确。

目的：观察电针对脑缺血再灌注损伤大鼠Toll样受体4/核因子κB信号通路及炎性因子表达的影响。
方法：48只雄性SD大鼠随机分为假手术组、模型组和电针组，每组16只，采用大脑中动脉线栓阻断法制备脑缺血再灌注损伤大鼠模型。造模24 h后对电针组大鼠行电针干预，每天1次，每次20 min，共5 d；假手术组和模型组不做任何干预。干预5 d后根据Longa法评定各组大鼠神经功能损伤程度；TTC染色和苏木精-伊红染色分别观察大鼠脑梗死体积及脑组织病理形态，荧光定量PCR和Western blot分别检测大脑皮质中Toll样受体4、核因子κB mRNA和蛋白表达；酶联免疫法检测血清中炎性因子白细胞介素6、白细胞介素18和肿瘤坏死因子α水平。

结果与结论：①与假手术组比较，模型组大鼠神经功能评分、血清白细胞介素6、白细胞介素18、肿瘤坏死因子α水平、大脑皮质Toll样受体4、核因子κB mRNA及蛋白表达水平明显升高(P < 0.01)；与模型组比较，电针组大鼠神经功能评分、大脑皮质Toll样受体4、核因子κB mRNA及蛋白表达，血清白细胞介素6、白细胞介素18、肿瘤坏死因子α水平明显降低(P < 0.05，P < 0.01)；②与假手术组比较，模型组大鼠脑梗死体积明显增加(P < 0.01)；与模型组比较，电针组大鼠脑梗死体积减小(P < 0.05)；③模型组神经元排列紊乱，部分神经细胞消失，胞核固缩，结构不完整；电针组大脑皮质神经元变性及神经细胞数量丢失程度均有不同程度减轻；④结果表明，电针能明显改善脑缺血再灌注损伤模型大鼠的神经行为学，减轻脑组织损伤，有效降低血清炎性因子水平，其机制可能与抑制Toll样受体4/核因子κB信号通路有关。

https://orcid.org/0000-0003-4127-3358(罗敷)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 脑缺血再灌注损伤, 电针, 炎症, Toll样受体4, 核因子κB

Abstract: BACKGROUND: Inflammation is one of the important factors that induce cerebral ischemia-reperfusion injury. Studies have shown that electroacupuncture can effectively reduce inflammation after ischemic stroke and improve the symptoms of neurological deficits, but the mechanism is not clear.
OBJECTIVE: To observe the effect of electroacupuncture on Toll-like receptor 4/nuclear factor-κB in rats with cerebral ischemia-reperfusion injury.
METHODS: Forty-eight male Sprague-Dawley rats were randomly divided into sham operation group, model group and electroacupuncture group, with 16 rats in each group. The rat model of cerebral ischemia-reperfusion injury was prepared by middle cerebral artery occlusion. At 24 hours after modeling, the rats in the electroacupuncture group were treated with electroacupuncture, once a day, 20 minutes each time, for a total of 5 days. The sham operation group and the model group did not do any intervention. After 5 days of intervention, Longa method was used to evaluate the degree of neurological injury in rats. Triphenyl tetrazolium chloride staining and hematoxylin-eosin staining were used to measure the volume of cerebral infarction and the pathological changes of brain tissue in rats. Serum interleukin-6, interleukin-18 and tumor necrosis factor-α were detected by ELISA. Expressions of Toll-like receptor 4 and nuclear factor-κB in the cerebral cortex at mRNA and protein levels were detected by fluorescence quantitative PCR and western blot, respectively. 
Results and conclusion: Compared with the sham operation group, the neurological function scores, serum interleukin-6, interleukin-18, and tumor necrosis factor-α levels, Toll-like receptor 4 and nuclear factor-κB mRNA and protein expression levels were significantly higher in the model group (P < 0.01). Compared with the model group, electroacupuncture significantly reduced the neurological function scores, serum interleukin-6, interleukin-18, and tumor necrosis factor-α levels, Toll-like receptor 4 and nuclear factor-κB mRNA and protein expression levels (P < 0.05, P < 0.01). Compared with the sham operation group, the volume of cerebral infarction in the model group increased significantly (P < 0.01). Compared with the model group, the volume of cerebral infarction in the electroacupuncture group decreased (P < 0.05). In the model group, the arrangement of neurons was disordered, some nerve cells disappeared, nuclei presented with pyknosis and incomplete structure. After electroacupuncture intervention, the degree of neuronal degeneration and neuronal loss in the cerebral cortex of rats were reduced compared with those in the model group. To conclude, electroacupuncture can significantly improve the neurobehavior of rats with cerebral ischemia-reperfusion injury, reduce brain tissue injury, and effectively reduce the level of serum inflammatory factors. The mechanism may be related to the inhibition of Toll-like receptor 4/nuclear factor-κB signaling pathway. 

Key words: cerebral ischemia-reperfusion injury, electroacupuncture, inflammation, Toll-like receptor 4, nuclear factor-κB

中图分类号: