α7尼古丁受体基因抑制对SH-SY5Y细胞α-分泌酶水平的影响

Influence of inhibited α7 nicotinic acetylcholine receptor gene expression on level of α-secretase in SH-SY5Y Cells

目的采用RNA干扰技术抑制神经母细胞瘤细胞(SH-SY5Y细胞)中α7神经型尼古丁乙酰胆碱受体(Neuronal nicotinic acetylcholine receptor,nAChR)基因表达,了解对APP代谢中的关键酶α-分泌酶水平的影响。方法 SH-SY5Y细胞转染针对α7 nAChR的小干扰RNA(small interference RNA,siRNA),用Real-time PCR法和Western blotting法分别测定细胞中α7 nAChR、α-分泌酶(ADAM10)在mRNA和蛋白表达水平的变化。结果转染α7 nAChR siRNA后,与对照组相比,α7 nAChR mRNA及蛋白表达水平明显降低;ADAM10mRNA及蛋白表达水平分别减少了73%和66%。结论α7 nAChR基因表达抑制后α-分泌酶水平明显降低,这种改变可能与α7 nAChR基因表达抑制后β-淀粉样蛋白(Aβ)的产生增多有关,可能与阿尔茨海默病(AD)的发病有一定的关系。

Objective To investigate the influence of inhibited α7 neuronal nicotinic acetylcholine receptor（nAChR）by small interference RNA（siRNA）on the level of α-secratase,the key enzyme in APP metabolism,in SH-SY5Y cells.Method The siRNA of α7 nAChR was transfected into SH-SY5Y cells,and the expressions of α7 nAChR and α-secretases（ADAM10）at mRNA and protein levels were studied by real-time PCR and Western blotting,respectively.Result As compared with controls,the expression of α7 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with the α7 nAChR siRNA were decreased significnatly.The expressions of ADAM10 were decreased by 73%and 66%,respectively.Conclusion The inhibited α7 nAChR mRNA induced by siRNA may obviously decrease the level of α-secretase which may connected to the increased production of Aβ induced by the inhibiting of α7 nAChR,and might be connnected to the pathogenesis of AD.