解剖学报 ›› 2013, Vol. 44 ›› Issue (6 ): 756-760.doi: 10.3969/j.issn.0529-1356.2013.06.005

• 神经生物学 • 上一篇    下一篇

大鼠脊髓损伤后神经细胞凋亡及相关调控蛋白Caspase-3表达的时空分布

刁尧1*  刘新宁1,2  葛瑞3  焦英3  朱悦3   

  1. 1.中国医科大学实验技术中心,沈阳 110001; 2.阜新市第二人民医院神经内科,辽宁 阜新 123000;3. 中国医科大学附属第一医院骨科,沈阳 110001
  • 收稿日期:2012-10-09 修回日期:2012-11-14 出版日期:2013-12-06 发布日期:2013-12-06
  • 通讯作者: 刁尧 E-mail:diaoyao@yahoo.com
  • 基金资助:

    省自然科学基金资助项目;自然科学基金资助项目

Temporal and spatial expression pattern of apoptosis and its associated protein in neurons after spinal cord injury in rats

DIAO Yao 1* LIU Xin-ning 1,2 GE Rui3 JIAO Ying3 ZHU Yue3   

  1. 1. Center of Experimental and Technology, China Medical University, Shenyang 110001,China;2. Department of Neurology, the Second people’s Hospital, Liaoning Fuxin 123000,China; 3.Department of Orthopedics,the First Affiliated Hospital, China Medical University, Shenyang 110001,China
  • Received:2012-10-09 Revised:2012-11-14 Online:2013-12-06 Published:2013-12-06
  • Contact: DIAO Yao E-mail:diaoyao@yahoo.com

摘要:

目的 观察大鼠脊髓损伤(SCI)后神经元细胞凋亡和相关调控蛋白Caspase-3表达的时间、空间分布规律。方法 SD大鼠50只,分为假手术组和SCI组,钳夹法建立SCI模型,用结晶紫(CV)染色法、TUNEL法和免疫组织化学方法观察SCI后6h、12h、24h、3d和7d损伤中心到头端0~5.00mm空间范围内脊髓神经细胞存活、凋亡以及相关调控蛋白Caspase-3的表达状况。 结果 CV染色发现,SCI后6h~7d在0~0.50mm空间范围内均未发现存活的神经细胞,在1.00~3.50mm距离内,存活的神经细胞数逐渐增加,4.00mm处达峰值;TUNEL结果发现,SCI后6h~7d在1.05~4.55mm范围内,神经细胞出现凋亡,3d时在2.55mm处,细胞凋亡达高峰,7d时显著减少;免疫组织化学结果发现,SCI后6h~7d在1.10~4.60mm范围内,神经元细胞出现Caspase-3阳性表达,3d在3.10mm处Caspase-3表达达到高峰,7d显著减少。 结论 SCI后,凋亡的神经细胞及其相关调控蛋白Caspase-3的表达有一定的时空分布规律。

关键词: 脊髓损伤, 凋亡, 半胱氨酸天冬氨酸蛋白酶3, 原位缺口末端标记, 大鼠

Abstract:

Objective The purpose of this study is to evaluate temporal and spatial expression pattern of neuron apoptosis and its associated protein in neurons after spinal cord injury in rats. Methods Fifty SD rats were divided into a shamoperated and a spinal cord injury group, 25 rats per group. SCI model was made by clamps methods. The expression profiles were evaluated by counting the cresyl violet-stained surviving cells and the number of TUNEL-positive neurons. Caspase-3 expression was determined by immunohistochemical analysis in sections 0-5.00 mm rostral to the epicenter at 6,12,24 hours, 3 and 7 days after injury. Results Neurons continued to disappear over 7 days after injury and neuronal loss extended to the areas far away from the epicenter over times except 0-0.50mm. TUNEL-positive and Caspase-3 positive neurons in gray matter appeared after 6 hours, gradually increased to a peak level after 3 days at 2.55mm and 3.10mm, and declined by 7 days postinjury in the section 1.10-4.60mm rostral to the epicenter. Conclusion Temporal and spatial distribution of neurons apoptosis and Caspase-3 protein expression after spinal cord injury are regulated.

Key words: Spinal cord injury, Apoptosis, Caspase-3, TUNEL, Rat

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