摘要
目的:通过切断及切断后再修复坐骨神经,观察在其相应脊髓节段中Nogo-A表达的变化规律,了解在周围神经损伤及修复过程中相应中枢部位Nogo-A的作用规律,为合理的选择周围神经损伤的修复方法和修复时间,有效避免Nogo-A对周围神经损伤修复的影响提供基础理论依据。
方法:选用健康Sprague Dawley(SD)大鼠150只,体重约200克,雌雄不拘,按照实验目的随机分为6组,每组25只。第一组:切断坐骨神经。第二组:切断坐骨神经,随即在手术显微镜下用10/0无创伤缝合线缝合神经外膜以修复切断的坐骨神经。第三组:切除坐骨神经1cm。第四组:切除坐骨神经1cm,随即用第三组切除之坐骨神经行同种异体神经移植。第五组:切除坐骨神经1cm,随即将切除之坐骨神经原位缝合神经外膜修复坐骨神经,模拟自体神经移植。第六组(假手术对照组):参照以上各组手术方法显露坐骨神经但不损伤。以上各组大鼠术后每一组随机分为5组,每组5只,分别于术后24小时、48小时、1周、2周、32天处死。经左心室—升主动脉插管灌注固定,然后迅速取出相应节段脊髓,将取出的脊髓多聚甲醛固定、脱水、浸蜡、包埋、切片、免疫组织化学处理,光镜观察,并对脊髓前角运动神经元行光密度测定。
结果:光镜下可见,Nogo-A在脊髓前角运动神经元胞浆内有明显
坐骨神经损伤后Nog,A在相应脊翻节段的表达
表达,而细胞核内未见表达。平均光密度测定显示,坐骨神经切断、
切除、切除后自体神经移植组及异体神经移植组脊髓灰质前角坐骨神
经损伤侧Nogo一A表达阳性的运动神经元平均光密度较未损伤侧明显
增高。其中,坐骨神经切断、切除及切除后自体神经移植组,相应损
伤侧脊髓前角Nogo一A表达阳性的运动神经元平均光密度增强的规律
基本相似,于伤后1、2天即出现增高,l周时达高峰,之后逐渐降低,
32天时趋向正常。坐骨神经切断后立即修复组,相应损伤侧脊髓前角
Nogo一A表达阳性的运动神经元平均光密度较未损伤侧无明显增强,且
各观察时间点平均光密度无明显差别、而坐骨神经切除后行异体神经
移植组,术后第1、2天相应损伤侧脊髓前角Nog。一A表达阳性的运动
神经元平均光密度即较未损伤侧明显增高,1周时增高幅度减小,2周
时达高峰,后虽逐渐降低,但至32天时仍处于较高水平。
结论:坐骨神经损伤后,脊髓相应节段损伤侧前角运动神经元胞
浆内Nogo一A的表达较未损伤侧明显增强。伤后立即行端一端吻合术修
复,脊髓相应节段损伤侧前角运动神经元胞浆内Nog。一A的表达基本
正常,无明显表达增强改变。伤后立即行自体神经移植术,脊髓相应
节段损伤侧前角运动神经元胞浆内Nogo一A的表达变化规律与不治疗
组相同;而立即行异体神经移植术,早期即可加重Nogo一A表达的增强,
在术后32天Nogo一A表达的增强仍维持于较高水平。
Objectives: To investigate the regularity of the expression of Nogo-A in spinal cord corresponding to traumatic sciatic nerve in the course of injury and repair.
Methods: 150 healthy adult Sprague Dawley(SD)rats were randomly divided into 6 groups, 25 per group. Group 1: sciatic nerves were severed without repair; Group 2: sciatic nerves were cut followed by immediate anastomosis of nerval adventitia using 10/0 non-traumatic suture; Group 3: 1 cm of sciatic nerve was resected in every subjects; Group 4: 1 cm resection of sciatic nerve was replaced by the resected sciatic nerve in group 3; Group 5: 1 cm of sciatic nerve was severed followed by epineurial suture in situ; Group 6: Visualization of sciatic nerve was performed without injury to it. The rats of each group were randomly divided into 5 subgroups, 5 per subgroup. The five subgroups were sacrificed at different intervals, 1 day, 2 day, 1 week, 2 week, 32day after operation. The expression of Nogo-A in spinal cord were detected by immunohistochemical staining and quantified by mean optical density (MOD).
Results: The expression of Nogo-A was positive in the cytochylema of motoneuron in anterior gray column of spinal cord. The MOD of traumatic side was evidently higher than non-traumatic side in group 1, 3, 5 at 1, 2 day after operation. The MOD of the severed side reached the top at 1 week.and then descented gradually, and almost normal at 32
days after operation. The MOD of traumatic side was not statistically different from non-traumatic side in the group 2. The MOD of traumatic side raised remarkable at 1 ,2 day after operation in the group of altogeneic transplantation, descented a little at 1 week, reach top at 2 week and maintained high level afterwards.
Conclusions: The expression of Nogo-A in motoneuron on traumatic side was higher than that on non-traumatic side. The expression of Nogo-A in traumatic side was different from non-traumatic side and came to normal on the condition that the traumatic nerve was immediately repaired by end-end anastomosis after injury. If the the traumatic nerve was replaced by allogeneic transplantation, the expression of Nogo-A in traumatic side were remarkable higher than that in non-traumatic side, and still maintained high level even after 1 month.
引文
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