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《Journal of Guiyang Medical College》 2006-01
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The Inhibitory Interference of RNA to α7 Neuronal Acetylcholic Receptor Gene Expression in SH-SY5Y Cells

QI Xiao-lan,GUAN Zhi-zhong(Molecular Biological Key Laboratory of Guiyang Medical College,Guiyang 550004,China)  
Objective: To inhibit α7 neuronal acetylcholic receptor(α7 nAChR) gene expression in human neuroblastoma cells and study the effects of inhibited expression of α7 nAChR on cell MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reduction and the level of lipid peroxidation.Methods: The α7 nAChR gene specific siRNAs were designed and synthesized by molecular biological method,and transfected into neuroblastoma cells.The cells were incubated for 48 hours before harvesting.The α7 nAChR mRNA and protein levels inside were monitored with RT-PCR and western blotting.For the measuring of lipid peroxidation and MTT reduction,the transfected neuroblastoma cells were treated with 1μmol/L Aβ_(25-35),and then,spectrophotometry and MTT method were used respectively.Results: Comparing with control group,in experimental group,α7 nAChR mRNA and protein levels decreased with the inhibitory rates of 52% and 36% respectively,and the product of lipid peroxidation increased,and MTT decreased.The α7 nAChR siRNA also increased the toxicity of Aβ_(25-35) on the cultured cells.Conclusions: The α7 nAChR siRNA can specifically inhibit the expression of α7 nAChR gene in neuroblastoma cells.Inhibition of α7 nAChR gene expression can enhance the toxicity of Aβ.α7 nAChR plays an important role in the neuroprotective function and its expression inhibiton may be connected to the pathogenesis of Alzheimer disease.
【Fund】: 国家自然科学基金资助项目(30460045)
【CateGory Index】: R749.161
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