egcg与锌离子抑制缺氧复氧损伤诱导的大鼠心肌细胞凋亡及相关机制word格式论文.docx

关键词缺血再灌注损伤；凋亡；PI3K/Akt；没食子儿茶素没食子酸；锌离子AbstractBackground and objective1、Apoptosisisoneofthemostimportantcharacteristicsofmyocardialischemiareperfusioninjury.TheactivationofPI3K-Aktpathwayhasbeenreganizedasoneofthemostsignificantmechanismsinmyocardialischemiareperfusion.2、ItwasreportedthatEGCGprotectedINS-1cellagainstoxidativestressbytheenhancementofanti-apoptosissignalingthroughtheincreasedlevelofphosphorylatedPI3K andAkt.However,thepropertiesofEGCGonPI3K/Aktpathwayinthemodelofmyocardialischemia-reperfusioninjuryhavenotbeenwellestablished.3、SeveralreportsrevealedthatexogenouszincprotectedthecardiomyocyetsagainstH/R-inducedapoptosisbytargetingthePI3k/Aktpathway.Meanwhile,zinccouldenhancethehepatoprotectivityofEGCGinisolatedrathepatocytes.WhethertheinteractionsofEGCGandzinchavecardiopretectioninresponsetomyocardialischemiareperfusioninjuryarestillunknown.Therefore,thebiochemicalmechanismunderlyingthecytoprotectionwasinvestigatedby examiningcellviabilitychangesat0μM、5μM、10μM、15μM、20μMofEGCGandZn2+intheH/RmodelofH9c2cells.WesoughttoinvestigatewhethertreatmentofEGCGandZn2+modulatedH/R-inducedapoptosis.MethodsInordertoestablishtheH/Rmodel,H9c2cellswereexposedtohypoxia(95%N2and5% CO2)for3hfollowedbyreoxygenationfor1hinnormoxic(37°C,95%air+5%CO2).WeperformedMTTassaystodeterminethedifferentconcentrationsofEGCG（0μM、5μM、10μM、15μM、20μM）and Zn2+（0μM、5μM、10μM、15μM、20μM）onthegrowthofH9c2cells.Accordingtothepreviousresults,wechosetheoptimizedconcentrationsof10μMEGCGand5μMZn2+,theseconcentrationswereusedforallfurtherexperiments.Thecellsweretrypsinized, plated(10000-12000cells/cm2)inculturedishes,thenwererandomlydividedintosixgroupsafterculturingfor48h:①normalgroup:withoutanytreatment;②H/Rgroup:hypoxiaandreoxygenationonly;③H/R+EGCGgroup:10μMEGCGwasadded30minbeforehypoxiaandreoxygenation;④H/R+Zn2+ group:5μMZn2+ wasadded30minbeforehypoxiaandreoxygenation;⑤H/R+EGCG+Zn2+group:10μMEGCG+5μMZn2+wasadded30min beforehypoxiaandreoxygenation;⑥H/R+EGCG+Zn2++LY294002group:10μMEGCG+5μMZn2++20μMLY29