神经损伤诱导面神经运动神经元死亡及caspase相关蛋白的表达

上海口腔医学 ›› 2015, Vol. 24 ›› Issue (1): 23-29.

神经损伤诱导面神经运动神经元死亡及caspase相关蛋白的表达

魏海刚¹,李蜀光¹,陈玉婷¹,蔡超雄¹,许彪²

1.广东省佛山市第二人民医院,广东佛山 528000;
2.昆明医科大学附属口腔医院,云南昆明 650031

收稿日期:2013-12-15 出版日期:2015-02-20 发布日期:2015-07-24
通讯作者: 李蜀光,E-mail:lishuguang1964@126.com
作者简介:魏海刚 (1974-),男,硕士,副主任医师,E-mail:whgqy437@sina.com
基金资助:
广东省医学科学技术研究基金(B2003144); 佛山市科技局医学专项基金(200508062）; 佛山市科技创新平台项目（2013AG10012）

Facial motoneurons death and caspase regulatory gene expression following facial nerve injury

WEI Hai-gang¹,LI Shu-guang¹,CHEN Yu-ting¹,CAI Chao-xiong¹,XU Biao²

1.Foshan Second People’s Hospital. Foshan 528000, Guangdong Province;
2.The Hospital of Stomatology of Kunming Medical University. Kunming 650031, Yunnan Province, China

Received:2013-12-15 Online:2015-02-20 Published:2015-07-24
Supported by:
Medical Science and Technology Research Foundation of Guangdong Province (B2003144), Medical Fund of Science and Technology Bureau of Foshan City (200508062) and Science and Technology Innovation Special Projects of Foshan City (2013AG10012)

摘要/Abstract

摘要： 目的研究大鼠面神经压榨伤及低位切断伤后,面神经运动神经元的形态学改变和死亡相关基因caspase 3、caspase 8、cyto-c的表达变化及其相关性。方法手术制作大鼠右侧面神经的压榨伤及低位切断伤模型,左侧为正常对照侧。用甲苯胺蓝染色及透射电镜技术观测面神经运动神经元形态学变化及死亡情况。免疫组织化学（S-P）法、图像分析技术检测切断伤及压榨伤后caspase 3、8和cyto-c的表达变化。采用SPSS10.0软件包对数据进行统计学分析。结果① 面神经切断伤及压榨伤均可引起面运动神经元死亡,且于伤后4周时达到高峰,切断伤组死亡数显著多于压榨伤组。死亡形式以凋亡为主。② caspase 3、caspase 8、cyto-c蛋白表达阳性神经元分布于正常面神经核各亚核,部分胶质细胞也有染色。切断伤组损伤侧细胞染色重于压榨伤组。损伤后3d时各蛋白表达开始增强,caspase 3、8表达于伤后14 d而cyto-c则于伤后7 d时达到高峰。Caspase 8、cyto-c蛋白表达与caspase 3蛋白表达相关。结论①不同形式神经损伤均可引起面神经运动神经元死亡,死亡数目与损伤形式相关。② 损伤后caspase 3、8和cyto-c蛋白表达变化与面神经损伤形式以及损伤时间有关,caspase 8、cyto-c表达与caspase 3表达相关。提示caspase 8、cyto-c可能参与激活caspase 3的过程。Caspase 级联反应在面神经运动神经元凋亡过程中有重要作用。

关键词: 面神经, 损伤, 细胞死亡, 调控基因, Caspase

Abstract: PURPOSE: To investigate the morphology of facial motoneurons and its death course as well as caspase 3, caspase 8, cyto-c death proteins’ expression and their correlation following facial nerve distal transection or crush in rats. METHODS: The right facial nerve underwent distal transaction and crush as experimental group, while the left facial nerve acted as normal control. We observed the morphology and the death course of motoneurons by light microscope and transmission electron microscope. Expressions of caspase 3, 8, and cyto-c protein were studied by immunohistochemistry (S-P) and image analysis. SPSS 10.0 software package was used for statistical analysis. RESULTS: ① Both axon distal transection and axon crush resulted in death of facial motoneurons. The motoneurons’ loss reached peak 28 days after injuries and were mainly through apoptotic pathway. The number of motoneurons’ loss in the distal transection group were more than that in the crush group. ② Caspase 3, caspase 8 and cyto-c protein expressions were observed in wide spread areas of normal rat facial nucleus. In addition to neurons, glial cells were also stained. Cells of the distal transection group stained more strongly than that of crush group. Expressions of the proteins began to increase 3 days after the injuries. Caspase 3 and caspase 8 protein expression reached peak 14 days whereas cyto-c protein expression reached peak 7 days after the injuries. Expression of caspase 8 and protein were correlated with expression of caspase 3 protein. CONCLUSIONS: ①Different facial nerve injuries result in death of facial motoneurons. The loss of motoneurons is related with the injuries’ patterns. Clinical nerve reparation should be performed as early as possible within 4 weeks after the transection. ②The expression of caspase 3, 8 and cyto-c protein were related with facial nerve injuries’ patterns. Caspase 8 and cyto-c protein expressions were correlated with caspase 3 protein expression, indicating that caspase 8 and cyto-c may take part in activating caspase 3. Caspase cascade reaction may have an important role in the apoptosis of facial motoneurons.

Key words: Facial nerve, Injuries, Cell death, Regulatory genes, Caspase

中图分类号:

R739.8

魏海刚,李蜀光,陈玉婷,蔡超雄,许彪. 神经损伤诱导面神经运动神经元死亡及caspase相关蛋白的表达[J]. 上海口腔医学, 2015, 24(1): 23-29.

WEI Hai-gang,LI Shu-guang,CHEN Yu-ting,CAI Chao-xiong,XU Biao. Facial motoneurons death and caspase regulatory gene expression following facial nerve injury[J]. Shanghai Journal of Stomatology, 2015, 24(1): 23-29.

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