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ID:32211610
大小:4.63 MB
页数:35页
时间:2019-02-01
《小分子蛋白sulfiredoxin-1对大鼠星形胶质细胞氧化应激损伤后细胞凋亡的保护作用与 机制分析》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、万方数据重庆医科大学硕士研宄生学位论文SULFIREDoXIN.1AMELIoR√气TESoXIDArrIVESTRESSIN叫RYINDUCEDBYoXYGEN.GLUCoSEDEPRIV盯IoNANDHYDRoGENPEROXIDEINRArCoRTICALASTROCYTESVIAANTI.APoPTOSISBackground:ABSTRACTAstrocytesapoptosisplaysavitalroleinCNSinjuries,suchascerebralischemia,Alzheimer’Sdiseas
2、e,andParkinson’Sdisease.Sulfiredoxin.1(Srxn1),acentralendogenousantioxidantprotein,hasimportantneuroprotectivefunctionsincerebralischemiaincludingantioxidationandanti.apoptosis.However,themechanismsofSrxnlduringapoptosisincerebralischemiahavenotbeenwellstudied.Obje
3、ctive:TodetermineheroleofSrxnlinprotectionfromapoptosisinducedbyOGDorH202anditspossiblemechanism.Methods:Weusedlentiviralvector-encodingshorthairpinRNAs(shRNAs)totransfecttoastrocyteswithinterferenceSrxnl,andthenexposedthemtoOGDfor6hor100gMH202for6htOmodeloxidative
4、stressinjury.(2)WemeasuredcellviabilityandcelldamageasdeterminedbyMTSandLDHassays.万方数据重庆医科大学硕士研究生学位论文(3)UsingimmunocytochemistryHoechst33342andflowcytometryAV/PIdoubledyetodetectcellapoptosis.(4)JC一1probeandFluo一3,AMprobewereusedtodetectthechangeofmitochondrialmemb
5、ranepotentialandintracellularfreeGa2+usingLaserconfocal(5)UsingWBtomeasuretheproteinofSrxnl,cyt.C,caspase一9,caspase-3,PARRbax,bcl-2,andcaspase一12;andQPCRtomeasurethemRNAlevelofSrxnl.Results:(1)TheresultsofMTSandLDHshowedthatinterferenceSrxnlattenuatedcellviabilitya
6、ndincreasedcelldamage(p<0.05,p<0.01).(2)Hoechst33342andFCMshowedthatinterferenceSrxnlincreasedapoptosis(p<0.O1).(3)JC-1stainingshowedthatinterferenceSrxnlinduceddissipationof△、
7、,mp8、3,caspase-9,PARRbax,anddeclinedantiapoptoticbcl-2proteinasdeterminedbyWB(p<0。05,p<0.01).Buttherewasnoobviouschangeincaspase-12proteinafterinterferenceSrxnlp>O.05)(5)Fluo一3,AMstainingshowedthatafterinterferenceSrxnlthelevelofintracellularCa。werenotdifferentfromtheco9、ntrolgroup(p>0.05)6万方数据重庆医科大学硕士研究生学位论文Conclusion:TheseresultssuggestedthatSrxnlcanprotectprimaryratcorticalastrocytesfromOGD·-orH202·-inducedcell
8、3,caspase-9,PARRbax,anddeclinedantiapoptoticbcl-2proteinasdeterminedbyWB(p<0。05,p<0.01).Buttherewasnoobviouschangeincaspase-12proteinafterinterferenceSrxnlp>O.05)(5)Fluo一3,AMstainingshowedthatafterinterferenceSrxnlthelevelofintracellularCa。werenotdifferentfromtheco
9、ntrolgroup(p>0.05)6万方数据重庆医科大学硕士研究生学位论文Conclusion:TheseresultssuggestedthatSrxnlcanprotectprimaryratcorticalastrocytesfromOGD·-orH202·-inducedcell
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