解剖学报

解剖学报 ›› 2014, Vol. 45 ›› Issue (1): 20-25.doi: 10.3969/j.issn.0529-1356.2014.01.004

• 神经生物学 • 上一篇    下一篇

表没食子儿茶素没食子酸酯对大鼠脊髓损伤后神经细胞凋亡及Caspase-3 表达的影响

刘新宁1,2 刁尧1* 兰振和2 马婧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 1100015.
  • 收稿日期:2013-01-10 修回日期:2013-02-16 出版日期:2014-02-06 发布日期:2014-02-06
  • 通讯作者: 刁尧 E-mail:diaoyao@yahoo.com
  • 基金资助:

    No:20092118;省自然科学基金资助项目;自然科学基金资助项目

Effect of epigallocatechin gallate on temporal and spatial profiles of neurons apoptosis and Caspase-3 protein expression after spinal cord injury in rats

LIU Xin-ning 1,2 DIAO Yao 1* LAN Zhen-he2 MA Jing2 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:2013-01-10 Revised:2013-02-16 Online:2014-02-06 Published:2014-02-06
  • Contact: DIAO Yao E-mail:diaoyao@yahoo.com

摘要：

目的 观察表没食子儿茶素没食子酸酯（EGCG）对脊髓损伤(SCI)大鼠神经元凋亡和凋亡调控蛋白Caspase-3时间、空间表达的影响，探讨其脊髓损伤保护机制。
方法 SD大鼠75只，分为假手术组、SCI组、EGCG组（n=25只）；钳夹法制作脊髓损伤模型，EGCG组和SCI组损伤后即刻和1h后腹腔注射EGCG（50 mg/kg）和等量生理盐水；用结晶紫（CV）染色法、TUNEL和免疫组织化学方法，观察SCI后6h、12h、24h、3d和7d时间范围内，从损伤中心到头端0～5.00mm空间范围内，EGCG对脊髓神经元存活、凋亡以及相关调控蛋白Caspase-3表达的影响。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元，在1.00～4.60mm范围内，EGCG和SCI组相比存活的神经元数明显增加（P＜0.01）；TUNEL结果发现，SCI后6h～7d，在1.05～3.05mm范围内, EGCG与SCI组相比凋亡神经元数明显减少（P＜0.01）；免疫组织化学发现，SCI后6h～7d，在1.10～3.10mm范围内，EGCG与SCI组相比Caspase-3阳性表达的神经元数明显减少（P＜0.01）。结论 EGCG在一定时间、空间范围内能下调Caspase-3表达，减少神经元凋亡，对继发性SCI有拮抗作用。

关键词: 脊髓损伤, 表没食子儿茶素没食子酸酯, 凋亡, Caspase-3, 原位缺口末端标记, 大鼠

Abstract:

Objective To evaluate the effect of epigallocatechin gallate(EGCG) on temporal and spatial profiles of neuron apoptosis and its associated Caaspase-3 protein expression after spinal cord injury in rats. Methods SD rats were randomly divided into three groups (25 rats each): sham-operated group, SCI group (normal saline solution intraperitoneal injection, immediately and 1 hour after SCI), and EGCG treatment groups (50mg/kg, intraperitoneal injection, immediately and 1 hour after SCI ). SCI model was made by clamps methods. The rats were sacrificed at 6, 12, 24 hours, 3 and 7days; 5 rats at each time point after injury. Spinal cord tissue in the injured area was harvested, sectioned 0-5.00 mm rostral from the epicenter post-injury and examined by cresyl violet (CV) staining and terminal-deoxynucleoitidyl transferase (TdT)-mediated deoxy uridine triphosphate-biotin nick end labeling (TUNEL) staining method and immunohistochemical method. Results At all the time points observed, CV-staining result showed that the neurons was not found in section 0-0.50 mm for SCI and EGCG group, the neurons was increased in EGCG group than in the SCI group in the sections 1.00-4.60 mm. The number of TUNEL positive neurons and Caspase-3 neurons were decreased in EGCG group than in SCI group in the section 1.05-3.05 mm and 1.10-3.10 mm rostral from the epicenter. Conclusion EGCG can decrease the expression of Caspase-3 and inhibit neuronal cell apoptosis in the rats after spinal cord injury on temporal and spatial profiles, which may play a significant neuroprotective role in the secondary SCI.

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