中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (5): 671-676.doi: 10.3969/j.issn.2095-4344.2016.05.011

• 周围神经损伤动物模型 Animal models of peripheral nerve injury • 上一篇    下一篇

急性马尾神经压迫后脊髓前角运动神经元细胞凋亡相关蛋白的表达规律

王 展1，李浩鹏2，贺西京2，姬 刚3，张 军1，年跃文1，张 堃1   

  1. 1西安交通大学医学院附属红会医院，陕西省西安市 710054；2西安交通大学附属第二医院骨科，陕西省西安市 710004；3西安市中心医院骨外科，陕西省西安市 710054
  • 收稿日期:2015-11-03 出版日期:2016-01-29 发布日期:2016-01-29
  • 通讯作者: 张堃，主任医师，西安交通大学医学院附属红会医院，陕西省西安市 710054
  • 作者简介:王展，男，陕西省西安市人，西安交通大学医学院毕业，硕士，副主任医师，主要从事脊柱与脊髓损伤、创伤骨科、矫形方面的研究。
  • 基金资助:
    国家自然科学基金面上资助项目(30271310)

Expression of apoptosis-related protein in motor neurons of anterior horn of the spinal cord after acute cauda equina compression

Wang Zhan1, Li Hao-peng2, He Xi-jing2, Ji Gang3, Zhang Jun1, Nian Yue-wen1, Zhang Kun1   

  1. 1Xi’an Honghui Hospital, Health Science Center, Xi’an Jiaotong University, Xi’an 710054, Shaanxi Province, China; 2Department of Orthopedics, Second Hospital, Xi’an Jiaotong University, Xi’an 710004, Shaanxi Province, China; 3Department of Bone Surgery, Xi’an Central Hospital, Xi’an 410054, Shaanxi Province, China
  • Received:2015-11-03 Online:2016-01-29 Published:2016-01-29
  • Contact: Zhang Kun, Chief physician, Xi’an Honghui Hospital, Health Science Center, Xi’an Jiaotong University, Xi’an 710054, Shaanxi Province, China
  • About author:Wang Zhan, Master, Associate chief physician, Xi’an Honghui Hospital, Health Science Center, Xi’an Jiaotong University, Xi’an 710054, Shaanxi Province, China
  • Supported by:

     the General Program of National Natural Science Foundation of China, No. 30271310

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863

被引次数

4

摘要：

文章快速阅读：

文题释义：

急性马尾神经压迫：常继发于腰椎骨折或腰椎间盘突出，即使在8 h以内手术彻底解除马尾神经压迫，也往往遗留有较多的后遗症状，包括会阴区皮肤感觉减退，尿便控制功能降低，甚至出现男性功能受损。马尾神经纤维受压后通过负反馈作用可能会导致脊髓前角运动神经元的病变，也许是马尾综合征疗效不尽人意的根本所在。
细胞凋亡：是一个主动过程，它涉及一系列蛋白的激活、表达以及调控等的作用。其中Caspase家族蛋白、Bcl-2家族蛋白、Bax等在调亡的信号转导中扮演重要角色。

 

背景：马尾综合征往往遗留有会阴区皮肤感觉减退，尿便控制功能降低，甚至出现男性功能受损。实验研究发现外周神经纤维损伤后，神经元胞体会发生凋亡，这与损伤的性质、神经元种类、动物种属、年龄、与神经元距离等因素有关。
目的：探讨急性马尾神经压迫后脊髓前角运动神经元细胞凋亡及相关蛋白的表达规律。
方法：27只家犬随机分为3组，压迫组及对照组制备马尾神经压迫模型，正常组不造模。压迫组又分为水囊压迫持续4，8，12，24，48，72，168 h组，每组3只；对照组只置入水囊，不注水。采用TUNEL标记法检测脊髓前角运动神经元凋亡情况，免疫组化SABC法检测 Bax、Bcl-2、Caspase-3蛋白的表达，用Qwin550Cw型图像采集与分析系统检测 Bax、Bcl-2、Caspase-3蛋白表达阳性细胞灰度值。
结果与结论：急性马尾神经压迫后脊髓前角运动神经元细胞发生凋亡，压迫12 h即有阳性细胞检出，72 h神经元凋亡达到高峰。Bax、Bcl-2蛋白在正常组即有少量表达，Caspase-3蛋白在正常组、对照组无表达。Bax、Bcl-2蛋白表达于压迫持续8 h开始显著增加，72 h达高峰，168 h降至正常；Bax蛋白表达增幅较Bcl-2大。Caspase-3蛋白于压迫持续12 h开始表达，72 h达高峰，168 h仍有少量表达；Bax、Caspase-3蛋白表达在72 h达高峰，Bcl-2蛋白表达则无明显上升。提示急性马尾神经压迫后脊髓前角运动神经元发生凋亡；Bax、Bcl-2蛋白表达呈相互拮抗作用，在Bax/Bcl-2复合体中，Bax蛋白占优势时促进凋亡，诱发Caspase-3蛋白表达，导致神经元凋亡发生。 

ORCID: 0000-0002-8010-455X(王展)

关键词: 实验动物, 神经损伤与修复动物模型, 马尾神经, 急性压迫, 脊髓前角运动神经元, 凋亡, 凋亡相关蛋白, Bax, Bcl-2, 国家自然科学基金

Abstract:

BACKGROUND: Cauda equina syndrome often induces skin hypoesthesia in the perineal area, poor urine-stool control, and impairs male function. After peripheral nerve fiber injury, apoptosis of neurons appeared. This is associated with the nature of the injury, the types of neurons, the species of animals, the age, and the distance between neurons.
OBJECTIVE: To explore the motor neuron apoptosis and expression of apoptosis-associated protein in the anterior horn of the spinal cord after acute cauda equina compression. 
METHODS: A total of 27 canines were randomly divided into three groups. In the compression and control groups, models of cauda equina compression were established. In the normal group, no models were established. Compression group received water sac compression for 4, 8, 12, 24, 48, 72 and 168 hours, with three models in each group. In the control group, only water sac was implanted, but water was not injected. Terminal deoxynucleotidyl transferase TdT-mediated biotin dUTP nick end-labeling assay was used to detect the apoptosis of neurons in the anterior horn of the spinal cord. Bcl-2, Bax and Caspase-3 protein expressions were measured by immunohistochemical staining (strept avidin-biotin complex). Gray values of positive cells of Bax, Bcl-2 and Caspase-3 protein expressions were detected using Qwin550Cw image collection and analysis system.
RESULTS AND CONCLUSION: The apoptosis of motor neuron occurred in the compression groups. At 12 hours of compression, positive cells were detected, and the number of positive cells reached a peak at 72 hours. Bax and Bcl-2 protein expression was small in the normal group. Caspase-3 protein expression was not detected in the normal and control groups. Bax and Bcl-2 protein expression was significantly increased at 8 hours, peaked at 72 hours and reduced to a normal level at 168 hours. The increased range of Bax protein expression was bigger than that of Bcl-2. Caspase-3 protein began to express at 12 hours, peaked at 72 hours and reduced to a low level at 168 hours. Bax and Caspase-3 protein expression peaked at 72 hours, and Bcl-2 protein expression was not obviously increased. These findings verified that after acute cauda equina compression, the apoptosis of neurons occurred in the anterior horn of the spinal cord. Bax and Bcl-2 protein expression showed an antagonistic action. In the Bax/Bcl-2 complex, Bax protein in a high expression promoted apoptosis, induced Caspase-3 protein expression, and neuronal apoptosis.