机构地区:[1]海南医学院人体解剖学教研室,海口市571199
出 处:《中国脊柱脊髓杂志》2016年第5期441-446,共6页Chinese Journal of Spine and Spinal Cord
基 金:海南省自然科学资金(814299);海南医学院科研培育资金(HY2010-008);海南省教育厅科研项目(Hjkj2013-32)
摘 要:目的 :探讨脊神经前根损伤后,大鼠脊髓组织中Slit1的表达变化,为进一步研究Slit1在神经再生中的作用提供依据。方法:应用2月龄SD大鼠共100只,体重250±20g,其中80只SD大鼠实施左侧L5及L6脊神经前根切断,分别在伤后1d、3d、7d、14d时处死(每个时间点20只),取L5~L6脊髓节段,标记左右;左侧半L5~L6脊髓节段为实验组;右侧半为自身对照组。假手术大鼠(20只)实施麻醉及暴露L5及L6脊神经手术,未行L5及L6脊神经前根切断术,取其L5~L6脊髓节段为空白对照组。采用免疫组化、Western blotting及RT-PCR法检测大鼠脊髓组织中Slit1的变化。结果:脊神经前根切断后1d、3d、7d、14d,实验组中Slit1阳性细胞数分别为5.78±1.53、15.85±2.65、23.93±1.53、8.78±1.78;自身对照组中分别为2.31±1.63、2.57±1.89、3.20±2.16、3.02±2.15。各时间点实验组中Slit1阳性细胞数显著高于空白对照组(2.89±1.26)及自身对照组(P〈0.05)。Western blotting示脊神经前根切断后1d、3d、7d、14d,实验组Slit1相对表达量分别为0.326±0.09、0.448±0.05、0.638±0.07、0.304±0.07;自身对照组分别为0.038±0.02、0.038±0.01、0.035±0.02、0.046±0.03。各时间点实验组Slit1相对表达量与空白对照组(0.038±0.03)及自身对照组比较,差异有显著性(P〈0.05)。RT-PCR示脊神经前根切断后1d、3d、7d、14d,实验组Slit1 m RNA相对量分别为0.380±0.03、0.425±0.04、0.768±0.05、0.605±0.04;自身对照组分别为0.210±0.04、0.265±0.03、0.292±0.02、0.261±0.02。各时间点实验组Slit1 m RNA相对量与空白对照组(0.231±0.03)及自身对照组比较,差异有显著性(P〈0.05)。结论 :脊神经前根切断后,同侧脊髓灰质前角内Slit1的表达显著增加。Objectives: To investigate the changes of Slit1 expression in spinal cord after the section of dorsal root in rats, and to provide evidence for the role of Slit1 in nerve regeneration. Methods: The left dorsal roots of L5 and L6 spinal nerve were transected from 80 2-month-old Sprague-Dawley rats, and the changes of Slit1 expression were detected with RT-PCR, Western blotting and immunohistochemical methods in the related spinal cord at 1d, 3d, 7d and 14 d, respectively. The left halves of these L5/6 spinal cords were included in test group, the right halves of these L5/6 spinal cords were included in self-control group and the L5/6spinal cords from sham operation rats(which were anaesthetized and the L5 and L6 spinal nerves were exposed, but without section operation, n =20) in blank control group. Then the number of Slit1 positive cells and the average integral optical density value(AIODV) were determined with image analysis. Results: The numbers of Slit1 positive cells in test group at 1d, 3d, 7d and 14 d were 5.78±1.53, 15.85±2.65, 23.93±1.53 and 8.78±1.78; the numbers in self-control group were 2.31±1.63, 2.57±1.89, 3.20±2.16 and 3.02±2.15, respectively. The number of Slit1 positive cells in test group was significantly higher than that in blank control groups(2.89 ±1.26) or that in self-control group(P 0.05). Western blotting showed the relative expressions of Slit1 in test group at 1d, 3d, 7d and 14 d were 0.326 ±0.09, 0.448 ±0.05, 0.638 ±0.07 and 0.304 ±0.07; the numbers in self-control group were 0.038 ±0.02, 0.038 ±0.01, 0.035 ±0.02 and 0.046 ±0.03 respectively. Compared with blank control group(0.038±0.03) and self-control group, the relative expression of Slit1 in test group were significant(P〈0.05). RT-PCR showed the relative expressions of Slit1 m RNA in test group at 1d,3d, 7d and 14 d were 0.380±0.03, 0.425±0.04, 0.768±0.05 and 0.605±0.04; the numbers in self-control group were 0.210±0.04, 0.265±0.03, 0.292±0.02 and 0.261±0.02 respectively.
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