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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 observe the effect of epigallocatechin-3-gallate (EGCG) on the neuronal apoptosis and the expression of Caspase-3 in the spinal cord during spinal cord injury (SCI). Methods 75 Sprague-Dawley rats were randomly divided into sham operation group, SCI group and EGCG group (n = 25). Spinal cord injury models were made by clamp method. EGCG and 50 mg / kg) and the same amount of normal saline. The time from injury center to the tip 0 ~ 5.00mm was observed within 6h, 12h, 24h, 3d and 7d after SCI by crystal violet (CV) staining, TUNEL and immunohistochemistry Effect of EGCG on Survival, Apoptosis and Caspase-3 Expression in Spinal Cord Neurons in Spatial Area. Results CV staining showed that no surviving neurons were found in the spatial scale of 0 ~ 0.50mm after 6h ~ 7d after SCI. In the range of 1.00 ~ 4.60mm, the number of surviving neurons in EGCG and SCI groups increased significantly (P <0.01). The results of TUNEL showed that the number of apoptotic neurons in EGCG and SCI groups was significantly decreased from 6h to 7d after SCI (1.05-3.05mm) (P <0.01) Compared with SCI group, the number of Caspase-3-positive neurons in EGCG significantly decreased in the range of 1.10-3.10mm (P <0.01). Conclusions EGCG can down-regulate the expression of Caspase-3 and decrease the apoptosis of neurons within a certain time and space, and antagonize the secondary SCI.