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目的评价Nogo-A基因在视神经损伤后修复机制中的作用。方法Nogo-A基因敲除联合视神经损伤鼠为实验组(20只),C57BL/6小鼠联合视神经损伤作为对照组(20只)。制备视网膜、视神经冰冻切片,采用免疫荧光技术检测视网膜神经节细胞和视神经中生长相关蛋白(GAP)-43的表达。进行视网膜神经节细胞体外培养,进行GAP-43染色,采用图像分析系统计算细胞轴突长度。结果视神经中GAP-43表达:夹伤后1、3、7d对照组中表达少量GAP-43,实验组中GAP-43表达明显高于对照组(t=2·12,3·56、2·63,均P<0·01)。GAP-43抗体染色可见着色在体外培养RGC轴突,实验组3d有较长轴突生长,对照组3d有较短轴突生长。RGC于培养1、3及7d,经自动图像分析系统处理,得出细胞突起长度,实验组突起长度明显高于对照组(F=41·36、31·23,均P<0·01)。结论Nogo-A基因在抑制视神经损伤后轴突再生机制中起重要作用。
Objective To evaluate the role of Nogo-A gene in the repair mechanism of optic nerve injury. Methods Nogo-A gene knockout combined optic nerve injury rats were experimental group (20 rats), C57BL / 6 mice combined with optic nerve injury as control group (20 rats). The retina and optic nerve were frozen sectioned, and the expression of growth associated protein (GAP) -43 in retinal ganglion cells and optic nerve was detected by immunofluorescence technique. Retinal ganglion cells were cultured in vitro and stained with GAP-43. Cell axon length was calculated using an image analysis system. Results GAP-43 expression in the optic nerve: A small amount of GAP-43 was expressed in the control group at 1, 3 and 7 days after wounding, and the expression of GAP-43 in the experimental group was significantly higher than that in the control group (t = 2 · 12,3 · 56,2 · 63, all P <0.01). GAP-43 antibody staining staining RGC axons cultured in vitro, the experimental group had longer axons growth 3d, control group 3d short axons growth. RGCs cultured for 1, 3, and 7 days were processed by automatic image analysis system to obtain the cell protrusion length. The length of protrusion in the experimental group was significantly higher than that in the control group (F = 41.36,31.23, P <0.01). Conclusion Nogo-A gene plays an important role in inhibiting axonal regeneration after optic nerve injury.