解剖学报

解剖学报 ›› 2024, Vol. 55 ›› Issue (2): 143-149.doi: 10.16098/j.issn.0529-1356.2024.02.003

• 神经生物学 • 上一篇    下一篇

脱细胞异体神经移植物联合电针对坐骨神经损伤大鼠脊神经节的保护作用及机制

周泽宇1 马蕴涵1 李佳瑞1 胡余梦1 袁博1 张银娟2 于晓敏3 付秀美1,4*   

  1. 1.承德医学院基础医学院人体解剖学教研室，河北 承德067000; 2.承德医学院中医系针灸推拿教研室，河北 承德 067000; 3.承德医学院教务处，河北 承德067000; 4.河北省神经损伤与修复重点实验室，河北 承德067000
  • 收稿日期:2023-01-28 修回日期:2023-03-02 出版日期:2024-04-06 发布日期:2024-04-06
  • 通讯作者: 付秀美 E-mail:fuxiumei2012@163.com
  • 基金资助:
    施万细胞样细胞来源的NGF通过PI3K/Akt信号通路促进周围神经再生修复的分子机制;施万样细胞来源的BDNF调控大鼠坐骨神经再生修复的分子机制

Protective effect and mechanism of acellular nerve allografts combined with electroacupuncture on spinal ganglia in rats with sciatic nerve injury

ZHOU  Ze-yu1  MA  Yun-han1  LI  Jia-rui1  HU  Yu-meng1  YUAN  Bo1  ZHANG  Yin-juan2  YU  Xiao-min3  FU  Xiu-mei1,4*   

  1. 1.Department of Human Anatomy, College of Basic Medicine, Chengde Medical College, Hebei Chengde 067000, China; 2.Department of Acupuncture and Massage, Traditional Chinese Medicine, Chengde Medical College, Hebei Chengde 067000, China; 3.Office of Academic Affairs, Chengde Medical College, Hebei Chengde 067000, China; 4.Hebei Key Laboratory of Nerve Injury and Repair, Hebei Chengde 067000, China
  • Received:2023-01-28 Revised:2023-03-02 Online:2024-04-06 Published:2024-04-06
  • Contact: FU Xiu-mei E-mail:fuxiumei2012@163.com

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摘要：

目的 探讨脱细胞异体神经移植物（ANA）联合电针对坐骨神经损伤（SNI）大鼠脊神经节的保护作用及机制。  方法  选取50只雄性SD大鼠，10只制备ANA；40只随机分为正常组、模型组、ANA组和联合组，每组10只。分离右侧坐骨神经，于梨状肌下缘5 mm处去除10 mm制备SNI模型。ANA组将制备好的ANA连接在损伤神经的两断端处。联合组于ANA连接术后2 d采用电子针疗仪电针“环跳穴”和“阳陵泉穴”，15 min/d，7 d为1个疗程，共4个疗程。电生理检测各组大鼠坐骨神经传导速度，评估受损轴突再生情况；尼氏染色观察脊神经节中神经元的形态结构；Western blotting和免疫荧光法检测脊神经节中神经生长因子（NGF）和脑源性神经营养因子（BDNF）的表达。  结果  与正常组比较，模型组大鼠坐骨神经传导速度明显降低（P<0.01）；神经节神经元中的尼氏体因肿胀、溶解导致结构不完整，数量显著减少（P<0.01）；NGF和BDNF蛋白表达量明显降低（P<0.01）。与模型组比较，ANA组和联合组大鼠坐骨神经传导速度明显增加（P<0.01）；神经节神经元中尼氏体损伤减弱，数量明显增加（P<0.01）；NGF和BDNF蛋白表达量显著升高（P<0.01）。与ANA组比较，联合组大鼠坐骨神经传导速度显著增加（P<0.01）；神经节神经元中尼氏体形态较规则，数量明显增加（P<0.01）；NGF蛋白表达量显著升高（P<0.01）。  结论  ANA联合电针可提高大鼠坐骨神经传导速度，改善神经节中神经元的形态结构，发挥对脊神经节神经元的保护作用，其机制可能与提高神经元中NGF和BDNF蛋白的表达，尤其是NGF蛋白的表达相关。

关键词: 脱细胞异体神经移植物, 电针, 脊神经节, 神经生长因子, 脑源性神经营养因子, 坐骨神经损伤, 免疫印迹法, 大鼠

Abstract:

Objective To investigate the protective effect and mechanism of acellular nerve allografts (ANA) combined with electroacupuncture on spinal ganglia in rats with sciatic nerve injury (SNI).   Methods Totally 50 male adult SD rats were randomly selected for this experiment. Ten rats were prepared for the ANA. Forty male SD rats were randomly divided into normal group, model group, ANA group and combinational group, with 10 rats in each group. The SNI model was established by cutting off the nerves 10 mm at the 5 mm on the inferior border of piriformis after separating the right sciatic nerves. The rats in the ANA group were bridged with ANA to the two broken ends of injured nerves. The rats in the combinational group were treated with electroacupuncture 2 days after ANA bridging, Huantiao (GB30) and Yanglingquan (GB34) were performed as the acupuncture points, each electroacupuncture lasted 15 minutes and 7 days as a course of treatment, 4 courses in all. Sciatic nerve conduction velocity was measured by electrophysiology to evaluate the regeneration of damaged axons. Morphology of spinal ganglia was observed by Nissl staining. The expression of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) were detected by Western blotting and immunofluorescent staining.   Results Compared with the normal group, the sciatic nerve conduction velocity in model group decreased significantly (P<0.01), Nissl bodies in neurons of spinal ganglia were swollen and dissolved, with incomplete structure and the number decreased dramatically (P<0.01), while the level of NGF and BDNF also decreased significantly(P<0.01). Compared with the model group, the sciatic nerve conduction velocity in ANA and combinational groups strongly increased (P<0.01), the damage of Nissl bodies in neurons of spinal ganglia reduced and the number obviously increased (P<0.01), the level of NGF and BDNF increased considerably (P<0.01). Compared with the ANA group, the sciatic nerve conduction velocity in combinational group increased significantly (P<0.01), the morphology of Nissl bodies in neurons of spinal ganglia were more regular and the number increased (P<0.01), moreover, the level of NGF also increased significantly (P<0.01).   Conclusion ANA combined with electroacupuncture can enhance the sciatic nerve conduction velocity, improve the morphology of neurons in spinal ganglia and play a protective effect on spinal ganglia. The mechanism can be related to the higher expression of NGF and BDNF proteins, especially the expression of NGF protein. 

Key words: Acellular nerve allograft, Electroacupuncture, Spinal ganglion, Nerve growth factor, Brainderived neurotrophic factor, Sciatic nerve injury, Western blotting, Rat 

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