大鼠脊髓损伤后不同时期轴突导向因子slit-2 mRNA表达及形态学变化
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国家自然科学基金资助项目(NO653BAO603)


Expression of slit-2 mRNA and morphology changes during the spinal cord hemisection injury of rats
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    目的:研究脊髓损伤后轴突导向因子slit-2的表达变化,同时观察大鼠行为学及脊髓形态学的变化特点-方法:40只成年SD大鼠随机分为正常组,假手术组和手术1-3-7-14-21-28天组,制作脊髓右侧半横断损伤模型,BBB法进行右下肢功能评分,HE染色观察损伤脊髓不同时期的形态学变化,原位杂交法观察slit-2 mRNA的表达变化,计算阳性细胞灰度值,并进行统计学分析-结果: 大鼠脊髓损伤后,肢体瘫痪从第7天开始明显恢复直至第28天;形态学显示了从局部血肿形成,到周围炎性细胞浸润再到损伤位点远端和近端的脊髓组织逐渐发生空泡变性的过程;原位杂交结果为正常组和假手术组脊髓前角运动细胞slit-2 mRNA的表达较低,脊髓损伤后第3天,损伤位点近端前角运动神经元中slit-2 mRNA的表达明显增加,第7天与第14天前角运动神经元中slit-2 mRNA的表达较前一时间组明显下降,并持续表达至28天,各组较假手术组表达明显增高,同时,第7天组损伤位点周围反应性增生的细胞中slit-2 mRNA的表达明显上升- 结论:成年哺乳动物脊髓损伤是一个动态变化的复杂过程,轴突导向因子slit-2可能通过介导炎性细胞的迁移及与炎性细胞分泌的若干介质和因子相互作用排斥新生轴突的生长,阻止其穿过损伤部位与远端重建突触联系;损伤切口周围反应性增生的细胞中slit-2 mRNA的表达上升提示了其可能参与损伤后的胶质瘢痕及抑制性微分子环境的形成-

    Abstract:

    Objective:To investigate the expression of axon guidance cue slit-2 mRNA, and the ethology and morphology changes in different phases of spinal cord injury(SCI) development. Methods:Forty Sprague-Dawley rats were randomly divided into normal group, sham operation group and operation group which include 6 subgroups(1,3,7,14,21,28 days). The spinal cord hemisection injury model was made by cutting the posterior right side spinal cord. Function changes in the right posterior limb were evaluated by BBB method, the general morphology changes were observed by HE stain, and the expression of slit-2 mRNA was detected through in situ hybridization methods. The positive cells were analyzed by a computer image analysis system. Results:The limb function of experimental rats significantly recovered from 7 d to 28 d. The morphology gradually changed from local hematoma to inflammatory cell infiltrate, scar formation and vacuolar degeneration of the distal as well as the proximal spinal cord tissue. The in situ hybridization results indicated that expression of slit-2 mRNA in the angulus anterior was low in either normal or sham operation group, but significantly increased at 3 d(P < 0.01) in the operation group, then it decreased significantly at 7 d and 14 d(P < 0.05), and continued expression until 28 d. Meanwhile, in 7 d group, the expression of slit-2 mRNA was obviously rising up in the reactive hyperplasia cells around the lesion incision. Conclusion:The injury of adult mammalian spinal cord is a dynamic complex course. The axon guidance cue slit may act as a inhibitory role to repel axon regeneration via mediating migration of inflammatory cells, interacting with several neurogen or cues, participating formation of the scar and other probable mechanisms.

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刘源,李文磊,丁新生,王京娥,朱东亚,江珺.大鼠脊髓损伤后不同时期轴突导向因子slit-2 mRNA表达及形态学变化[J].南京医科大学学报(自然科学版),2007,(7):680-683

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  • 收稿日期:2006-12-09
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