淀粉样前体蛋白和烟碱型乙酰胆碱受体α7亚型稳定共表达细胞模型的建立

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1、上海交通大学硕士学位论文淀粉样前体蛋白和烟碱型乙酰胆碱受体α7亚型稳定共表达细胞模型的建立姓名：沈颖华申请学位级别：硕士专业：药理学指导教师：殷明20070201上海交通大学硕士学位论文本工作构建了β淀粉样前体蛋白的真核表达载体并以之转染已稳定表达α7nAChR的SH-EP1，筛选蛋白稳定表达细胞株，从而构建了β淀粉样前体蛋白和α7nAChR稳定共表达的细胞模型，使独立研究α7nAChR激动在β淀粉样前体蛋白加工修饰中的作用机理成为可能，也为体外筛选高效能的烟碱受体亚型激动剂奠定了基础。关键词：阿尔茨海默病，烟碱型乙酰胆碱受体α7亚型，β淀粉样前体

2、蛋白，SH-EP1上海交通大学硕士学位论文CONSTRUCTIONOFCELLMODELSTABLYCO-EXPRESSINGAMYLOIDPRECURSORPROTEINANDα7NICOTINICACETYLCHOLINERECEPTORABSTRACTAlzheimer’sdisease(AD)isakindofneurodegenerationdisease,whichiscommonamongtheoldpeopleandischaracterizedbyprogressingdementia.Theaccumulationofbeta-

3、amyloidprotein(Aβ)isthoughttobethehallmarkandcontributemosttothegenerationofADwhichcanbesporadicorfamilial.MostofADcasesaresporadicwhichresultfromheterogeneouscauses,includinggeneticandenvironmentalfactors.ManystudiesinADresearchfocusontheeffectofgeneticfactorsonADpathogenesis

4、,revealingthatmutationcanaffecttheactivityofγ-secretasebywhichtheamyloidprecursorprotein(APP)canbeprocessedintoAβ.However,itisnotwellknownwhetherandhowtheenvironmentalandphysiologicalsignalscanregulatetheγ-secretaseactivityandAβproduction.Epidemiologicalstudieshaveshownthatthe

5、rateofADisrelativelylowamongthesmokingpeople,上海交通大学硕士学位论文suggestingthatactivatingthenicotinicacetylcholinereceptorhasprotectiveeffectonAD.Thenicotinicacetylcholinereceptor(nAChR)isakindofligandgatedionchannelamongthecellmembrane.Itplayssignificantroleinmaintainingtheintracellu

6、larcalciumandneurotransmitteracetylcholineatnormallevel.LatestevidencefrominvitroandinvivoresearchsuggeststhatthenAChRandAβarecloselyrelated.ButitisnotclearwhichofthenAChRsubtypecontributesmosttotheAβproductionandhowitworks.HerewebuildtheexpressionvectorincludingtargetedAPP695

7、geneandtransfectedtheSH-EP1cellswhichcanstablyexpressα7nAChR.WeselectedthetransfectedcelllinestoidentifystableexpressionofAPPandfinallybuildtheinvitrocellmodelwhichco-expressestheAPPaswellasα7nAChR,makingitpossibletoinvestigatetheselectiveeffectofnAChRsubtypeactivationonAPPint

8、racellularprocessing.Thismodelcanalsobeusedtoscreenthecompoun