Down Regulation of a Matrix Degrading Cysteine Protease Cathepsin L, by Acetaldehyde Role of CEBPα 英文参考文献.docVIP

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Down Regulation of a Matrix Degrading Cysteine Protease Cathepsin L, by Acetaldehyde Role of CEBPα 英文参考文献.doc

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Down Regulation of a Matrix Degrading Cysteine Protease Cathepsin L, by Acetaldehyde Role of CEBPα 英文参考文献

DownRegulationofaMatrixDegradingCysteine ProteaseCathepsinL,byAcetaldehyde:RoleofC/EBPa RiyazA.Mir,ShyamS.Chauhan* DepartmentofBiochemistry,AllIndiaInstituteofMedicalSciences,AnsariNagar,NewDelhi,India Abstract Background: The imbalance between extra cellular matrix (ECM) synthesis and degradation is critical aspect of various hepatic pathologies including alcohol induced liver fibrosis. This study was carried out to investigate the effect of acetaldehydeonexpressionofanextracellularmatrixdegradingproteasecathepsinL(CTSL)inHepG2cells. MethodologyandResults:Wemeasuredtheenzymaticactivity,proteinand,mRNAlevelsofCTSLinacetaldehydetreated anduntreatedcells.ThebindingofCAATenhancerbindingproteina(C/EBPa)toCTSLpromoteranditskeyroleinthe transcription from this promoter and conferring responsiveness to acetaldehyde was established by site directed mutagenesis, electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP) assays and siRNA technology. Acetaldehyde treatment significantly decreased CTSL activity and protein levels in HepG2 cells. A similar decreaseinthemRNAlevelsandpromoteractivitywasalsoobserved.Thisdecreasebyacetaldehydewasattributedtothe fallintheliverenrichedtranscriptionfactorC/EBPalevelsandit’sbindingtotheCTSLpromoter.MutagenesisofC/EBPa binding motifs revealed the key role of this factor in CTSL transcription as well as conferring responsiveness to acetaldehyde.ThesiRNAmediatedsilencingoftheC/EBPaexpressionmimickedtheeffectofacetaldehydeonCTSLlevels anditspromoteractivity.Italsoabolishedtheresponsivenessofthispromotertoacetaldehyde. Conclusion: Acetaldehyde down regulates the C/EBP a mediated CTSL expression in hepatic cell lines. The decreased expressionofCTSLmayatleastinpartcontributetoECMdepositioninliverwhichisahallmarkofalcoholicliverfibrosis. Citation:MirRA,ChauhanSS(2011)DownRegulationofaMatrixDegradingCysteineProteaseCathepsinL,byAcetaldehyde:RoleofC/EBPa.PLoSONE6(6): e20768.doi:10.1371/journal.pone.0020768 Editor:Christia