Generation of stable ARE- driven reporter system for monitoring oxidative stress

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1、Motaharietal.DARUJournalofPharmaceuticalSciences(2015)23:38DOI10.1186/s40199-015-0122-9RESEARCHOpenAccessGenerationofstableARE-drivenreportersystemformonitoringoxidativestress11*121*PariaMotahari,MajidSadeghizadeh,MehrdadBehmanesh,ShaghayeghSabriandFat

2、emehZolghadrAbstractBackground:NF-E2-relatedfactor2(Nrf2)-antioxidantresponseelement(ARE)signalingpathwayisthemajordefensivemechanismagainstoxidativestressandisupregulatedbyspecificantioxidantsandoxidantstocomprisethechemoptotectiveresponse.Detectionof

3、ARE-activatingcompoundshelpstodevelopnewdrugsandidentify/quantifythetensionrangeoftheoxidants.Importantreasonspromotingthisworkarehighthroughput,rapidandinexpensiveexperimentsrelativetotheinvitrostudiesforARE-Nrf2pathwaymonitoringofchemicalsandenvironm

4、entalsamples.Methods:InthisstudyhepatomaHuh7reportercelllinewasgeneratedwhichcontainsaluciferasegeneunderthecontrolofanARE.ThisisthefirstexampleofAREconstructcontainingonecopyofextendedconsensusresponseelement.Thecellsweretreatedwithhydroquinone(HQ)and

5、p-benzoquinone(BQ)(oxidativestressinducers)andtheantioxidant,curcumin.Results:Theluciferaseactivitywasinducedinaconcentration-dependentmannerinaconcentrationrangeof1–2μMforBQandHQ.CurcuminwasalsovalidatedasanAREinducerinconcentrationabove10μM.Inadditio

6、n,thisreportercelllineprovidesarapiddetectionasearlyas4htorespondtotheAREinducers.Conclusion:Itisapowerfultoolforthesensitiveandselectivescreeningofchemicals,drugsandenvironmentalsamplesfortheirantioxidantandoxidantactivities.Keywords:Antioxidants,Nrf2

7、-ARE,Oxidativestress,ReportercelllineBackgroundprotectthecells[3].MountingevidencedemonstratedthatMonitoringcompoundsonpathwaysofinteresttoprovidehumangenomeconsistsofprotectivegenesactingthroughinsightintothemolecularmodeofactiontoevaluatethetheenhanc

8、edsequenceknownastheAREintheirpro-possibletoxicityisrequiredinmodernmedicine.Further-moterregion.MetabolizationbytheAREmechanismismore,thelackofinformationaboutthecellularsignalingoneofthecommonresponses,whichisalsoinducedbythepathwayso