Nogo-A受体在早期神经元细胞分化过程中的表达

Expression of Nogo-A Receptors during Early Neuronal Differentiation Process

目的:探讨在早期神经元细胞生长发育过程中Nogo-A受体(NG-R)的表达变化。方法:体外培养PC12细胞,实验组加入50 ng/m L细胞生长因子(NGF)进行诱导分化1、3、5、7 d,对照组中不加入NGF诱导分化。于不同时间点镜下观察细胞轴突发育及细胞分化情况,免疫荧光法观察NG-R蛋白在PC12细胞中的表达与定位,RT-PCR法检测NG-R m RNA在PC12细胞中的表达变化,western blot法检测NG-R蛋白在PC12细胞中的表达变化。结果:随着诱导分化时间的增加,实验组PC12细胞轴突发育及细胞分化增加;对照组PC12细胞未检测出NG-R m RNA及蛋白表达;实验组随着NGF诱导刺激时间延长,PC12细胞内NG-R m RNA及蛋白表达量逐步增加,组间差异有统计学意义,且均高于对照组(P<0.05或0.01),但实验组诱导1 d PC12细胞内NG-R蛋白表达量与对照组差异无统计意义(P>0.05)。结论:在神经元发育早期NG-R的表达随着轴突生长逐渐升高。

Objective： To explore the expression of Nogo-A receptors（NG-R） during the early neuronal differentiation process. Methods： PC12 cells were cultured in vitro, and 50 ng/mL nerve growth factor（NGF） was added into medium to induce neuronal differentiation for 1, 3, 5, and 7 days individually in the experimental group but not in the control group. The cell differentiation and axon growth were observed by inverse microscopy. The expression of NG-R m RNA and protein in PC12 cells were detected by RT-PCR, western blot and immunofluorescence staining. Results： In the experimental group, the axon growth and differentiation of PC12 cells increased from day 1 to day 7. NG-R m RNA and protein were not detected in the control group. In the experimental group, the expression of NG-R m RNA and protein were increased from day 1 to day 7（P〈0.05 or 0.01）. But there was no difference of expression of NG-R protein at day 1 between the experimental and control groups（P 〉0.05）. Conclusion： The expression of NG-R increased during the early neuronal differentiation process.