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为检测大鼠肌源神经营养因子对体外培养大鼠脊髓运动神经元划痕损伤后 Bcl-2表达的影响 ,本实验从成鼠和乳鼠骨骼肌中提取出肌源神经营养因子。在胎鼠脊髓运动神经元培养第 7d时将其随机分成对照组和实验组 (成鼠组和乳鼠组 ) ,取 10 0μl肌源神经营养因子 ( 0 .1μg/ ml)加入实验组 ,对照组加入等量的 PBS,同时进行划痕损伤。损伤后第 3d计数各组存活的运动神经元 ,行 Nissl染色和免疫组化反应 ,计数 Bcl-2免疫反应 ( Bcl-2 -IR)阳性神经元数量 ,并在图像分析仪上对 Bcl-2 -IR阳性神经元作光密度分析。结果发现 ,损伤后第 3d,实验组运动神经元存活数 ,Bcl-2 -IR阳性神经元数和平均光密度均明显高于同期对照组 ,而乳鼠肌源神经营养因子组功效优于成鼠肌源神经营养因子组。结果提示 ,大鼠肌源神经营养因子对体外培养的受损运动神经元具有增强 Bcl-2表达、减少凋亡和损伤修复等作用 ,且乳鼠的此因子效能尤佳
In order to investigate the effect of myogenic neurotrophic factor (BDNF) on the expression of Bcl-2 in rat spinal cord motoneurons induced by scratch injury, myogenic neurotrophic factor was extracted from the skeletal muscle of adult and suckling mice. At the 7th day after culture, fetal motor neurons of fetal rat spinal cord were randomly divided into control group and experimental group (adult rats and neonatal rats), and 100 μl myogenic neurotrophic factor (0.1 μg / ml) was added to the experimental group. The same amount of PBS was added to the group while scratch damage was performed. The number of Bcl-2-IR positive neurons counted by Nissl staining and immunohistochemistry was counted on the 3rd day after injury, and the expression of Bcl-2 -IR positive neurons for optical density analysis. The results showed that on the 3rd day after injury, the number of motoneurons, the number of Bcl-2-IR positive neurons and the mean optical density in the experimental group were significantly higher than those in the control group, while the effect of the muscle-derived neurotrophic factor group Murine myelogenous neurotrophic factor group. The results suggest that the rat myogenic neurotrophic factor can enhance the expression of Bcl-2, reduce apoptosis and repair the damaged motor neurons cultured in vitro, and this factor is particularly effective in neonatal rats