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

Effects of α7 neuronal nicotinic receptor gene overexpression in SH-SY5Y cells on the levels of CaMK Ⅱ and CREB

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-pc DNA3.1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。

b 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.