目的:观察神经生长导向因子Slit2在大鼠急性脊髓损伤后表达变化和分布情况,探讨轴索再生导向机制。方法:成年SD大鼠用Allen’s方法制作脊髓挫伤(SCI)模型,分别于挫伤后第2,4,7,14天取材,半定量RT—PCR分析Slit2 mRNA的表达变化,免疫荧光激光共聚焦扫描观察Slit2蛋白在脊髓损伤区的分布。结果:RT—PCR显示SCI后出现Slit2转录上调,表达量持续增高并于伤后第7天达到顶峰,之后逐步下降;免疫荧光则显示Slit2阳性颗粒先后出现在脊髓损伤区灰质胶质细胞中及神经元胞膜上,以前角、中央管及中线周围和后角较为聚集。结论:成年大鼠脊髓急性损伤后存在Slit2表达一过性上调并在损伤区灰质内广泛分布,可能对再生轴索的导向性生长发挥重要作用。
Objective: To observe the Slit2 expression distribution and expression changes during spinal cord injury(SCI) of grown rats, and to investigate the molecular biological reason for the regenerative guidance of growth cone. Methods: The model of SCI was made with Allen's method. Seventy-two adult SD rats were randomly divided into SCI groups, sham operation group and normal control group. After the spinal cords of rat were injured, semi-quantitative reverse transcription polymerase chain reaction was applied to detect the expression changes of Slit2 mRNA; immunofluorescence was performed to detect the location of Slit2 protein expression in the gray matter. Results: The expression of Slit2 mRNA in damaged spinal cord appeared at the 2nd day post-trauma at a low level and increased significantly afterwards, which reached the peak at the 7th day post-trauma, then began to decrease during the following phase. The Slit2 immunofluorescence positive signal appeared at the extensive area of spinal cord gray, located in the astrocyte. Conclusion: This study demonstrates that Slit2 mRNA was expressed in the damaged local and vicinity part of gray matter, and it is likely to play a role in the axon regeneration after SCI.