RAGE Modulates HypoxiaReoxygenation Injury in Adult Murine Cardiomyocytes via JNK and GSK-3β Signaling Pathways 英文参考文献.docVIP

RAGEModulatesHypoxia/ReoxygenationInjuryinAdult MurineCardiomyocytesviaJNKandGSK-3bSignaling Pathways LinshanShang,RadhaAnanthakrishnan,QingLi,NosirudeenQuadri,MarianeAbdillahi,ZhengbinZhu, WuQu,RosaRosario,FatoumaToure′,ShiFangYan,AnnMarieSchmidt,RavichandranRamasamy* DivisionofSurgicalScience,DepartmentofSurgery,CollegeofPhysiciansandSurgeons,ColumbiaUniversity,NewYork,NewYork,UnitedStatesofAmerica Abstract Background: Advanced glycation end-products (AGEs) have been implicated in diverse pathological settings including diabetes,inflammationandacuteischemia/reperfusioninjuryintheheart.AGEsinteractwiththereceptorforAGEs(RAGE) andtransducesignalsthroughactivationofMAPKsandproapoptoticpathways.Inthecurrentstudy,adultcardiomyocytes werestudiedinaninvitroischemia/reperfusion(I/R)injurymodeltodelineatethemolecularmechanismsunderlyingRAGE- mediatedinjuryduetohypoxia/reoxygenation(H/R). Methodology/PrincipalFindings:Cardiomyocytesisolatedfromadultwild-type(WT),homozygousRAGE-null(RKO),and WTmicetreatedwithsolubleRAGE(sRAGE)weresubjectedtohypoxiafor30minutesaloneorfollowedbyreoxygenation for 1hour. In specific experiments, RAGE ligand carboxymethyllysine (CML)-AGE (termed ‘‘CML’’ in this manuscript) was evaluatedinvitro.LDH,amarkerofcellularinjury,wasassayedinthesupernatantinthepresenceorabsenceofsignaling inhibitor-treatedcardiomyocytes.CardiomyocytelevelsofheterogeneousAGEsweremeasuredusingELISA.Apronounced increaseinRAGEexpressionalongwithAGEswasobservedinH/Rvs.normoxiainWTcardiomyocytes.WTcardiomyocytes after H/R displayed increased LDH release compared to RKO or sRAGE-treated cardiomyocytes. Our results revealed significant increases in phospho-JNK in WT cardiomyocytes after H/R. In contrast, neither RKO nor sRAGE-treated cardiomyocytes exhibited increased phosphorylation of JNK after H/R stress. The impact of RAGE deletion on GSK-3b phosphorylationinthecardiomyocytessubjectedtoH/Rrevealedsignificantlyhigherlevelsofphospho-GSK-3b/totalGSK- 3b in RKO, as well as in s