α7尼古丁受体基因抑制对SH-SY5Y细胞胆碱酯酶及胆碱乙酰化酶水平的影响  

Influence of inhibited α7 nicotinic acetylcholine receptor gene expression on the levels of acetylcholinesterase and choline acetyltransferase in SH-SY5Y cells

作　　者：李毅[1] 王凡[2] 吴昌学[1] 官志忠[1,3] 齐晓岚[1] 

机构地区：[1]贵阳医学院分子生物学重点实验室,贵州贵阳550004 [2]贵阳医学院附属医院神经外科,贵州贵阳550004 [3]贵阳医学院病理学教研室及附院病理科,贵州贵阳550004

出　　处：《中风与神经疾病杂志》2013年第10期874-876,共3页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金资助项目(81360178);贵州省科技厅社会发展攻关项目[黔科合SY字(2013)3020号]及优秀青年人才项目[黔科合人字(2013)44号];贵州省教育厅优秀科技人才项目[黔教合KY字(2012)089号]

摘　　要：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(nAChR)基因表达,了解对细胞胆碱酯酶(AChE)及胆碱乙酰化酶(ChAT)水平的影响。方法 SH-SY5Y细胞转染针对α7 nAChR的小干扰RNA(siRNA),用real-time PCR法和Western blotting法分别测定细胞中α7nAChR、AChE及ChAT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,α7nAChR mRNA及蛋白表达水平明显降低;α7 nAChR基因表达抑制能明显升高AChE及ChAT mRNA和蛋白表达水平,尤其是AChE的表达水平。结论α7 nAChR基因表达抑制可能通过增加细胞AChE水平而增加神经递质乙酰胆碱的分解,而ChAT的表达升高可能为一种代偿机制,这可能与阿尔茨海默氏病(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 acetylcholinesterase（ACHE） and choline aeetyhransferase （CHAT） in SH-SY5Y cells. Method The siRNA of α7 nAChR was transfected into SH-SYSY cells, and the expressions of α7 nAChR, ChE and ChAT at mRNA and protein levels were mesured by real-time PCR and Western blotting, respectively. Result As compared with controls,the mRNA and protein levels of α7 nAChR in the SH-SYSY cells transfected with the α7 nAChR siRNA were signiflcnatly decreased. And the expressions of AChE and ChAT at mRNA and protein levels were increased,especially the AChE level. Conclusion The inhibited α7 nAChR mRNA induced by siRNA may obviously in- crease the level of AChE,which increase the acetylcholine decomposition and the increased expression of ChAT may be a compensatory mechanism, and might be connnected to the pathogenesis of AD.

关 键 词：α7尼古丁乙酰胆碱受体 胆碱酯酶 胆碱乙酰化酶 RNA干扰 

分 类 号：R749.16[医药卫生—神经病学与精神病学]