中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (11): 2252-2260.doi: 10.12307/2025.363

• 脊柱组织构建 spinal tissue construction • 上一篇    下一篇

重复性经脊髓磁刺激促进脊髓损伤小鼠运动功能恢复

宋海旺1，姜冠华1，穆盈盈2，付善羽2，孙宝飞1，李玉美1，余资江1，杨  丹1   

  1. 贵州医科大学，1基础医学院人体解剖学教研室，2基础医学院，贵州省贵安新区  561113


  • 收稿日期:2024-03-27 接受日期:2024-05-09 出版日期:2025-04-18 发布日期:2024-08-10
  • 通讯作者: 杨丹，博士，副教授，研究生导师，贵州医科大学基础医学院人体解剖学教研室，贵州省贵安新区 561113 共同通讯作者：余资江，博士，教授，研究生导师，贵州医科大学基础医学院人体解剖学教研室，贵州省贵安新区 561113
  • 作者简介:宋海旺，男，2000年生，贵州省思南县人，土家族，贵州医科大学在读硕士，主要从事神经系统损伤修复再生方面的研究。
  • 基金资助:
    贵州省科技计划项目(黔科合基础-ZK[2024]一般120，黔科合基础-ZK[2021]一般472)，项目负责人：杨丹；贵州省教育厅青年科技人才成长项目(黔科合KY字[2022]226号)，项目负责人：杨丹；贵州医科大学博士启动基金项目(校博合J字[2020]009号)，项目负责人：杨丹；贵州省卫生健康委员会科技基金项目(gzwkj-2021-509)，项目负责人：余资江

Repetitive trans-spinal magnetic stimulation promotes motor function recovery in mice after spinal cord injury

Song Haiwang1, Jiang Guanhua1, Mu Yingying2, Fu Shanyu2, Sun Baofei1, Li Yumei1, Yu Zijiang1, Yang Dan1   

  1. 1Human Anatomy Teaching and Research Laboratory, School of Basic Medical Sciences, Guizhou Medical University, Guian New District 561113, Guizhou Province, China; 2School of Basic Medical Sciences, Guizhou Medical University, Guian New District 561113, Guizhou Province, China
  • Received:2024-03-27 Accepted:2024-05-09 Online:2025-04-18 Published:2024-08-10
  • Contact: Yang Dan, MD, Associate professor, Master’s supervisor, Human Anatomy Teaching and Research Laboratory, School of Basic Medical Sciences, Guizhou Medical University, Guian New District 561113, Guizhou Province, China Co-corresponding author: Yu Zijiang, MD, Professor, Master’s supervisor, Human Anatomy Teaching and Research Laboratory, School of Basic Medical Sciences, Guizhou Medical University, Guian New District 561113, Guizhou Province, China
  • About author:Song Haiwang, Master candidate, Human Anatomy Teaching and Research Laboratory, School of Basic Medical Sciences, Guizhou Medical University, Guian New District 561113, Guizhou Province, China
  • Supported by:
    the Science and Technology Planning Foundation of Guizhou Province, Nos. QKH-ZK-[2024]-General 120 and QKH-ZK-[2021]-General 472 (to YD); the Technology Personnel Training Foundation of Guizhou Provincial Education Department, No. QKH-KY-[2022]-226 (to YD); the PhD Research Start-Up Foundation of Guizhou Medical University, No. XBH-J-[2020]-009 (to YD); the Guizhou Provincial Health Commission’ s Scientific and Technological Fund Project, No. gzwkj-2021-509 (to YZJ)

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摘要：


重复性经脊髓磁刺激(repetitive trans-spinal magnetic stimulation，rTSMS)：一种重复性脉冲磁场作用于局部脊髓节段，改变神经细胞的膜电位，使之产生感应电流，影响脊髓代谢和神经电活动，从而引起一系列生理生化反应的治疗技术。
Toll样受体4/核因子κB/NLRP3信号通路：Toll样受体4(Toll-like receptor 4，TLR4)活化后，促使核因子κB(nuclear factor kappa-B，NF-κB)进入细胞核，调控炎症相关基因的转录，引起NLRP3通路的过度激活。活化的NLRP3形成炎性小体，导致促炎因子的产生和炎症反应的进一步加剧。

背景：研究表明重复性经脊髓磁刺激能够抑制脊髓损伤炎症反应，在脊髓区域施加磁场刺激，以调节神经元的兴奋性和突触传递，从而促进神经系统的可塑性和修复。
目的：观察重复性经脊髓磁刺激对脊髓损伤后小鼠Toll样受体4/核因子κB/NLRP3信号通路的影响，探讨其对运动功能恢复的机制。
方法：将SPF级雄性C57BL/6J小鼠随机分为假手术组、脊髓损伤组、重复性经脊髓磁刺激组，后2组小鼠麻醉后采用咬骨钳剔除T9椎板，暴露脊髓，使用小型动脉瘤夹夹持脊髓20 s建立脊髓损伤模型。重复性经脊髓磁刺激组小鼠脊髓损伤的第1天开始进行为期21 d的重复性经脊髓磁刺激干预。每天持续10 min，每周刺激5 d，休息2 d。于脊髓损伤后1，3，7，14，21 d进行小鼠运动功能BMS评分，利用Western Blot检测脊髓损伤处的水通道蛋白AQP4、凋亡因子Bax、Bcl-2、CL-Caspase-3、炎症因子肿瘤坏死因子α、干扰素γ、白细胞介素6、白细胞介素4和Toll样受体4/核因子κB/NLRP3信号通路相关蛋白的表达，氧化应激试剂盒检测脊髓损伤处超氧化物歧化酶、谷胱甘肽过氧化物酶的活性及丙二醛浓度，免疫荧光染色法检测神经元核抗原的表达。
结果与结论：①重复性经脊髓磁刺激组小鼠的BMS评分高于脊髓损伤组(P < 0.05)；②与脊髓损伤组相比，重复性经脊髓磁刺激组脊髓含水量降低、AQP4蛋白表达降低；丙二醛浓度、Bax、CL-Caspase-3、肿瘤坏死因子α、干扰素γ、白细胞介素6与Toll样受体4/核因子κB/NLRP3信号通路相关蛋白的表达均降低(P < 0.05)，而超氧化物歧化酶活性、谷胱甘肽过氧化物酶活性、Bcl-2、白细胞介素4与神经元核抗原的表达均升高(P < 0.05)；③结果说明，重复性经脊髓磁刺激能下调Toll样受体4/核因子κB/NLRP3信号通路相关蛋白的表达，缓解脊髓损伤后的脊髓水肿、氧化应激、凋亡反应和炎症反应等，发挥神经保护作用，从而促进脊髓损伤后小鼠后肢运动功能的恢复。   

关键词: 脊髓损伤, 重复性经脊髓磁刺激, TLR4/NF-κB/NLRP3信号通路, 脊髓水肿, 氧化应激, 凋亡反应, 炎症反应, 神经保护, 运动功能

Abstract: BACKGROUND: Repetitive trans-spinal magnetic stimulation (rTSMS) can inhibit inflammatory responses following spinal cord injury. rTSMS applies magnetic field stimulation to the spinal cord region to modulate neuronal excitability and synaptic transmission, thereby promoting plasticity and repair of the nervous system.
OBJECTIVE: To observe the effects of rTSMS on the Toll-like receptor 4 (TLR4)/nuclear factor (NF)-κB/NLRP3 signaling pathway after spinal cord injury and explore its mechanism in promoting motor function recovery.
METHODS: Male C57BL/6J mice, SPF grade, were randomly divided into sham surgery group, spinal cord injury group, and rTSMS group. The latter two groups of mice were anesthetized and the T9 vertebral plate was removed using rongeur forceps to expose the spinal cord, and the spinal cord was clamped using a small aneurysm clip for 20 seconds to establish the spinal cord injury model. Mice in the rTSMS group underwent a 21-day rTSMS intervention starting on day 1 after spinal cord injury. The stimulation lasted 10 minutes per day, 5 days per week with an interval of 2 days. Basso Mouse Scale scores were used to assess motor function recovery in mice after spinal cord injury at 1, 3, 7, 14, and 21 days after spinal cord injury. Western blot was employed to detect the expression of AQP4, apoptotic factors Bax, Bcl-2, CL-Caspase-3, inflammatory factors tumor necrosis factor-α, interferon-γ, interleukin-6, interleukin-4, and the TLR4/NF-κB/NLRP3 signaling pathway related proteins in the injured spinal cord. Oxidative stress assay kit was used to measure the activity of superoxide dismutase, glutathione peroxidase, and malondialdehyde content at the site of spinal cord injury. Immunofluorescence staining was performed to detect the expression of neuronal nuclei (NeuN).
RESULTS AND CONCLUSION: The Basso Mouse Scale score in the rTSMS group was significantly higher than that in the spinal cord injury group (P < 0.05). 
Compared with the spinal cord injury group, the rTSMS group showed a reduction in spinal cord water content. The expression of AQP4 protein, malondialdehyde content, and expression of Bax, Bcl-2, CL-Caspase-3, tumor necrosis factor-α, interferon-γ, interleukin-6, and TLR4/NF-κB/NLRP3 signaling pathway related proteins were all decreased in the rTSMS group, while the activities of superoxide dismutase and glutathione peroxidase, as well as the expression of Bcl-2, interleukin-4, and NeuN, were all increased (P < 0.05). These results suggest that rTSMS downregulates the expression of proteins related to the TLR4/NF-κB/NLRP3 signaling pathway, alleviating symptoms after spinal cord injury such as spinal cord edema, oxidative stress, apoptosis, and inflammation, exerting neuroprotective effects, and thereby promoting the recovery of hindlimb motor function after spinal cord injury.

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

Key words: spinal cord injury, repetitive trans-spinal magnetic stimulation, TLR4/NF-κB/NLRP3 signaling pathway, spinal cord edema, oxidative stress, apoptosis, inflammation, neuroprotection, motor function

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