egcg对thp1源性泡沫细胞abca1影响及其机制word格式论文.docx

活化；另一方面，Keap1 从复合体上解离后，直接与IKKβ 结合，阻滞其下游对NF-κB的激活。结论：在THP-1源性泡沫细胞中，EGCG通过激活Nrf2/Keap1途径抑制TNF-α诱导的NF-κB激活，上调ABCA1的表达促进细胞内胆固醇流出。关键词：动脉粥样硬化，表没食子儿茶素没食子酸脂，ABCA1，Nrf2/Keap1途径，细胞核因子-κB。2Epigallocatechin-3-gallatepreventsTNF-α-induced NF-κB activation therebyup-regulatingABCA1 via Nrf2/Keap1 pathwayin macrophagefoamcellsABSTRACTBackgroundandobjective:ATP-bindingmembranecassettetransporterA1 (ABCA1)playsaprotectiveroleinthedevelopmentofatherosclerosisforthereverse cholesterol transportprocess. Epigallocatechin-3-gallate(EGCG),whichexists abundantlyingreentea,exertsananti-atheroscleroticeffectviaanti-inflammatory andmetabolicregulationactivities.Manygenesandproteinsrelatedtolipid metabolismareinvolvedintheloweringcholesteroleffectsofEGCG.However, effectsofEGCGonABCA1haverarelybeendescribed.Todemonstratetheeffectof EGCGonABCA1andpossiblemechanism,THP-1macrophagederivedfoamcells were exposedto TNF-α and establishingan inflammation cells condition.Methods:WefirstinducedTHP-1cellstobecomemacrophagesbyPMA.Then THP-1macrophageswereinducedtobefoamcellsbyox-LDL.Beforeexposureto TNF-α(10ng/ml),thefoamcellswerepretreatedbyEGCGwithdifferent concentrations(0,10,20,40,80μg/mL)andthencollectedforexperiments.Cellular cholesteroleffluxexperimentsandwereconductedtodeterminethecellularlipid accumulation.MTTweredeterminethecellstoxicity.mRNAlevelsofABCA1and LXRα/βwere determined byrealtimequantitativeRT-PCR.Luciferasereporterassay wereconductedtodetermineABCA1promoterlevels.Westernboltwereusedto determinetheprotein levels of ABCA1,LXRα/β,NF-κB p65, Nrf2, Keap1andIKKβ. TheDNA-combiningactivityofNF-κBandNrf2wereinvestigatedbyEMSA.Nrf2 and Keap1wereknockeddown bySmall interferingRNAtransfection.3Results:Inpresentstudies,wehavefoundthatexposureofmacrophagefoamcellsto TNF-αresultsinadown-regulationofABCA1andadecreaseincholesteroleffluxto apoA1,whichisattenuatedbypretreatmentwithEGCG.Moreover,ratherthan activatingtheLXRpathway,inhibitingtheTNF-α-inducedNF-κBactivitiesis detectedwithEGCGincells.Forinhi