Xu Kan, Chen Qi-xin, Li Fang-cai, Chen Wei-shan, Lin Min, Wu Qiong-hua
Department of Orthopaedic Surgery, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China.
J Zhejiang Univ Sci B. 2009 Mar;10(3):180-7. doi: 10.1631/jzus.B0820161.
To determine whether spinal cord decompression plays a role in neural cell apoptosis after spinal cord injury.
We used an animal model of compressive spinal cord injury with incomplete paraparesis to evaluate neural cell apoptosis after decompression. Apoptosis and cellular damage were assessed by staining with terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) and immunostaining for caspase-3, Bcl-2 and Bax.
Experiments were conducted in male Sprague-Dawley rats (n=78) weighing 300-400 g. The spinal cord was compressed posteriorly at T10 level using a custom-made screw for 6 h, 24 h or continuously, followed by decompression by removal of the screw. The rats were sacrificed on Day 1 or 3 or in Week 1 or 4 post-decompression. The spinal cord was removed en bloc and examined at lesion site, rostral site and caudal site (7.5 mm away from the lesion).
The numbers of TUNEL-positive cells were significantly lower at the site of decompression on Day 1, and also at the rostral and caudal sites between Day 3 and Week 4 post-decompression, compared with the persistently compressed group. The numbers of cells between Day 1 and Week 4 were immunoreactive to caspase-3 and B-cell lymphoma-2 (Bcl-2)-associated X-protein (Bax), but not to Bcl-2, correlated with those of TUNEL-positive cells.
Our results suggest that decompression reduces neural cell apoptosis following spinal cord injury.
确定脊髓减压在脊髓损伤后神经细胞凋亡中是否起作用。
我们使用了一种伴有不完全性截瘫的压迫性脊髓损伤动物模型来评估减压后的神经细胞凋亡。通过末端脱氧核苷酸转移酶(TdT)介导的脱氧尿苷三磷酸缺口末端标记(TUNEL)染色以及对半胱天冬酶-3、B细胞淋巴瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)进行免疫染色来评估凋亡和细胞损伤。
实验在体重300 - 400克的雄性Sprague-Dawley大鼠(n = 78)中进行。使用定制的螺钉在T10水平向后压迫脊髓6小时、24小时或持续压迫,随后通过移除螺钉进行减压。在减压后第1天、第3天或第1周、第4周处死大鼠。将脊髓整体取出,在损伤部位、损伤部位头侧和尾侧(距损伤7.5毫米处)进行检查。
与持续压迫组相比,减压后第1天减压部位以及减压后第3天至第4周损伤部位头侧和尾侧的TUNEL阳性细胞数量显著减少。第1天至第4周细胞对半胱天冬酶-3和Bcl-2相关X蛋白(Bax)呈免疫反应,但对Bcl-2无免疫反应,其数量与TUNEL阳性细胞数量相关。
我们的结果表明减压可减少脊髓损伤后的神经细胞凋亡。