SH－SY5Y 神经细胞中α7神经型尼古丁受体基因过表达对 CaMKⅡ和 CREB的影响

摘要

目的：研究 SH－SY5Y 神经细胞中α7 nAChR 基因过表达对 CaMKⅡ和 CREB的影响。方法复苏稳定转染α7 nAChR－pcDNA3．1质粒及空载质粒的 SH－SY5Y 神经细胞后，用含 G418的培养液进行筛选培养；应用实时荧光定量 PCR 法和蛋白印迹法（Western blot）检测α7 nAChR 基因过表达组、空载质粒组和正常对照组细胞中CaMKⅡ、CREB mRNA 及蛋白表达水平的变化。结果与对照组相比，α7 nAChR 基因过表达细胞组的 CaMKⅡ、CREB mRNA 表达水平分别增加了116．8％和114．7％（P ＜0．01）；及 CaMKⅡ、CREB 蛋白表达水平分别增加了8．7％（P ＞0．05）和41．4％（P ＜0．05）。结论α7 nAChR 的神经保护作用可能与上调细胞 CaMKⅡ、CREB 水平有关。%Objective To study the effects of α7 nAChR gene overexpression on the levels of CaMKⅡand CREB in SH-SY5Y cells.Method SH-SY5Y cells were transfected with α7 nAChR-pcDNA3.1 plasmid,the stable clones were cul-tivated with culture medium containing G418.Real-time PCR and Western blot were used to detect the mRNA and protein levels of CaMKⅡand CREB,respectively;Results Compared to negative controls and controls,SH-SY5Y with α7 nAChR gene overexpression increased the mRNA levels of CaMKⅡand CREB by 116.8% and 114.7%,respectively;increased the protein levels of CaMKⅡand CREB by 8.7% and 41.4%,respectively.Conclusion Neuroprotective effect of α7 nAChR may be related to up-regulation of CaMKⅡand CREB levels.