Redox Regulation of the AMP-Activated Protein Kinase 英文参考文献.docVIP

RedoxRegulationoftheAMP-ActivatedProteinKinase YingyingHan1,2,QilongWang1,PingSong1,YiZhu2,Ming-HuiZou1* 1Department of Biochemistry and Department of Medicine, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, United States of America, 2DepartmentofPhysiologyandPathophysiology,PekingUniversityHealthScienceCenter,Beijing,China Abstract Redoxstateisacriticaldeterminantofcellfunction,andanymajorimbalancescancauseseveredamageordeath. Objectives: The aim of this study is to determine if AMP-activated protein kinase (AMPK), a cellular energy sensor, is activatedbyoxidantsgeneratedbyBerberineinendothelialcells(EC). Methods:Bovineaorticendothelialcells(BAEC)wereexposedtoBerberine.AMPKactivityandreactiveoxygenspecieswere monitoredaftertheincubation. Results:InBAEC,Berberinecausedadose-andtime-dependentincreaseinthephosphorylationofAMPKatThr172and acetylCoAcarboxylase(ACC)atSer79,awellcharacterizeddownstreamtargetofAMPK.Concomitantly,Berberineincreased peroxynitrite,apotentoxidantformedbysimultaneousgenerationofsuperoxideandnitricoxide.Pre-incubationofBAEC with anti-oxidants markedly attenuated Berberine-enhanced phosphorylation of both AMPK and ACC. Consistently, adenoviral expression of superoxide dismutase and pretreatment of L-NG-Nitroarginine methyl ester (L-NAME; a non- selectiveNOSinhibitor)bluntedBerberine-inducedphosphorylationofAMPK.Furthermore,mitochondria-targetedtempol (mito-tempol) pretreatment or expression of uncoupling protein attenuated AMPK activation caused by Berberine. DepletionofmitochondriaabolishedtheeffectsofBerberineonAMPKinEC.Finally,Berberinesignificantlyincreasedthe phosphorylation of LKB1 at Ser307 and gene silencing of LKB1 attenuated Berberine-enhanced AMPK Thr172 phosphorylationinBAEC. Conclusion:Ourresultssuggestthatmitochondria-derivedsuperoxideanionsandperoxynitritearerequiredforBerberine- inducedAMPKactivationinendothelialcells. Citation: Han Y, Wang Q, Song P, Zhu Y, Zou M-H (2010) Redox Regulation of the AMP-Ac