Lin Yaofa, Zong Haiyang, Hu Xianteng, Yu Ronghua, Shao Wanwan, Hou Chunlin, Lin Haodong
Department of Orthopaedics, Shanghai Changzheng Hospital, the Second Military Medical University, Shanghai, 200040, P.R.China.
Department of Orthopaedics, Shanghai Changzheng Hospital, the Second Military Medical University, Shanghai, 200040,
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2017 Jan 15;31(1):80-84. doi: 10.7507/1002-1892.201610093.
To investigate the expression change of endogenous Spastin after sciatic nerve injury in rats, and to discuss the role and significance in the peripheral nerve regeneration.
Thirty-six adult male Sprague Dawley rats weighing 180-220 g were randomly divided into the experimental group ( =30) and the control group ( =6). Sciatic nerve compression damage model was established in the experimental group, and the sciatic nerve was only exposed in the control group. The L spinal cord tissue was obtained to detect Spastin mRNA and protein levels by real-time fluorescence quantitative PCR and Western blot at 1, 3, 7, 14, and 28 days after operation in the experimental group ( =6) and at 7 days in the control group. Meanwhile, the sciatic nerve at 5 mm distal to the injured site was obtained to observe the ultrastructure of the distal axon by transmission electron microscope (TEM).
The expression trends of Spastin gene and Spastin protein in L spinal cord tissue of 2 groups were basically identical. In the experimental group, the expressions of Spastin gene and protein decreased at the beginning, and then increased; the expressions reduced to the minimum at 7 days after operation, and came back to the initial level at 28 days. The expression levels of Spastin mRNA and protein at 3, 7, and 14 days were significantly lower in the experimental group than the control group ( <0.05), but no significant difference was noted between 2 groups at 1 and 28 days ( >0.05). The expression levels of Spastin mRNA and protein at 3, 7, and 14 days were significantly lower than those at 1 and 28 days in the experimental group ( <0.05), but no significant difference was noted between at 1 day and 28 days ( >0.05). At 1, 3, and 7 days after operation, the myelin damage was observed by TEM; at 14 days, there were regenerating Schwann cells; at 28 days, a large number of myelinated nerve fibers were seen, which were closed to normal form.
In the process of sciatic nerve regeneration after injury, a complex succession of changes take place in the expression of endogenous Spastin protein in rats, indicating that Spastin protein plays an important role in the process.
探讨大鼠坐骨神经损伤后内源性Spastin的表达变化，探讨其在周围神经再生中的作用及意义。
将36只体重180 - 220 g的成年雄性Sprague Dawley大鼠随机分为实验组（n = 30）和对照组（n = 6）。实验组建立坐骨神经压迫损伤模型，对照组仅暴露坐骨神经。实验组于术后1、3、7、14和28天（n = 6）及对照组于术后7天取L₄脊髓组织，采用实时荧光定量PCR和Western blot检测Spastin mRNA和蛋白水平。同时，取损伤部位远端5 mm处的坐骨神经，用透射电子显微镜（TEM）观察远端轴突的超微结构。
两组L₄脊髓组织中Spastin基因和Spastin蛋白的表达趋势基本一致。实验组中，Spastin基因和蛋白表达开始下降，随后上升；术后7天表达降至最低，28天恢复至初始水平。实验组术后3、7和14天Spastin mRNA和蛋白表达水平显著低于对照组（P < 0.05），但术后1天和28天两组间无显著差异（P > 0.05）。实验组术后3、7和14天Spastin mRNA和蛋白表达水平显著低于术后1天和28天（P < 0.05），但术后1天和28天之间无显著差异（P > 0.05）。术后1、3和7天，TEM观察到髓鞘损伤；14天，有再生的雪旺细胞；28天，可见大量有髓神经纤维，形态接近正常。
在大鼠坐骨神经损伤后的再生过程中，内源性Spastin蛋白表达发生复杂的连续变化，表明Spastin蛋白在此过程中起重要作用。
