NEP1-40促进高苯丙氨酸损伤大鼠神经元Nogo A表达

摘要

Objective The study is designed to investigate the effect by NEP1-40 on Nogo A in high phenyalanine (Phe) in vitro. Methods Cells was dissected from the cerebral cortex of El6 ～ El8 embryos of SD rat. Real-time PCR and Western blot were used to quantify mRNA and protein of Nogo A respectively. The growth cones, axon and the Nogo A location were performed by immunofluorescence and immunohistochemistry respectively. Results Compared to the control, the mRNA and protein level of Nogo A with Phe for 12 h, 24 h and 48 h were slightly reduced. NEP1-40 can upregulate expression of Nogo A on neuron in the high Phe condition with high level, but has no effect on normal neuron. Conclusion NEP1-40 can promote expression of Nogo A on neurons in high Phe,which facilitate the neuron grow.%目的 了解神经元NgR受体拮抗剂NEP1-40对高苯丙氨酸(Phe)作用下大鼠大脑皮质层神经元Nogo A蛋白表达的影响.方法 原代培养胎龄16～18 d的SD大鼠皮质神经元,体外模拟高Phe(0.9 mmol/L)环境,并用不同浓度NEP1-40干预;用免疫荧光方法检测神经元轴突生长和生长锥塌陷情况,实时荧光定量PCR和Western blot检测Nogo A mRNA和蛋白表达.结果 Nogo A可以在神经元胞体和突起表达;高Phe作用12、24和48 h神经元NogoA mRNA与α微管蛋白(α-Tub)的相对表达量明显减低(P<0.05);高Phe作用后神经元Nogo A蛋白明显降低(P<0.05);NEP1-40对正常神经元无促进作用,但对高phe作用下的神经元呈浓度依赖性地促进其Nogo A的表达.结论 NEP1-40可以促进高Phe损伤时神经元Nogo A的表达增高,从而促进神经元轴突的生长.