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。
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.
出处
《解剖学报》
CAS
CSCD
北大核心
2014年第1期20-25,共6页
Acta Anatomica Sinica
基金
辽宁省自然科学基金资助项目(20092118)
