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目的探讨NOGO-A在PC12细胞生长发育过程中的表达及意义。方法培养大鼠嗜铬细胞瘤细胞系PC12细胞,用神经生长因子诱导其分化,并于倒置显微镜下随机取20视野计数观察细胞增值和轴突生长情况。采用免疫荧光染色、逆转录酶聚合酶链反应(RT-PCR)及免疫印迹法等方法检测诱导后第1d、第3d、第5d、第7dPC12细胞中NOGO-A mRNA及蛋白的表达及变化,并留取细胞培养液检测多巴胺水平。结果未分化的PC12细胞中未检测到NOGO-A mRNA及蛋白表达。经神经生长因子诱导的PC12细胞,细胞轴突不断生长,NOGO-A mRNA及蛋白的表达逐渐增高(P<0.05)。PC12细胞在分化过程中多巴胺(DA)分泌水平无明显差别。结论PC12细胞向交感神经元分化的过程中NOGO-A的表达逐渐增强,推测NOGO-A在神经元发育早期可能促进轴突生长。但对多巴胺激素释放的调节不明显。
Objective To investigate the expression and significance of NOGO-A during the growth and development of PC12 cells. Methods The rat pheochromocytoma cell line PC12 was cultured and induced to differentiate by nerve growth factor. Cell proliferation and axon growth were observed under an inverted microscope. The expression and changes of NOGO-A mRNA and protein in PC12 cells on day 1, 3d, 5d and 7d after induction were detected by immunofluorescence staining, reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting, And keep cell culture medium to detect dopamine levels. Results NOGO-A mRNA and protein expression were not detected in undifferentiated PC12 cells. The expression of NOGO-A mRNA and protein in PC12 cells induced by nerve growth factor increased gradually (P <0.05). PC12 cells in the process of differentiation of dopamine (DA) secretion levels no significant difference. CONCLUSION: The expression of NOGO-A in PC12 cells gradually increases during sympathetic neuron differentiation. It is speculated that NOGO-A may promote axon growth in early neuronal development. However, the regulation of dopamine release is not obvious.