CombinationofAlcoholandCigaretteSmokeInducesEndoplasmicReticulumStressandCellDeathinPancreaticAcinarCellsAureliaLugea1,2,AndreasGerloff2,Hsin-YuanSu1,ZhihongXu4,ArielGo1,ChengHu1,SamuelW.French3,JeremyS.Wilson4,MinotiV.Apte4,RichardT.Waldron1,2,andStephenJ.Pandol1,21Cedars-SinaiMedicalCenter,LosAngeles,CA2VAGreaterLosAngelesHealthcareSystem/UniversityofCalifornia,LosAngeles,CA3Harbor-UCLAMedicalCenter,Torrance,CA4UniversityofNewSouthWales,Liverpool,AustraliaandInghamInstituteforAppliedMedicalResearch,Liverpool,AustraliaAbstractBackground&Aims—Smoking,anindependentriskfactorforpancreatitis,acceleratesthedevelopmentofalcoholicpancreatitis.AlcoholfeedingofmiceinducesupregulationofsplicedX-boxbindingprotein1(XBP1s),whichregulatestheendoplasmicreticulum(ER)unfoldedproteinresponseandpromotescellsurvivaluponERstress.WeexaminedwhethersmokingaffectstheadaptivemechanismsinducedbyalcoholandacceleratesdisordersoftheERinpancreaticacinarcells.Methods—Westudiedthecombinedeffectsofethanolandcigarettesmokeextract(CSE)onER-stressandcelldeathresponsesinmouseandhumanprimaryaciniandtheacinarcelllineAR42J.Cellswereincubatedwithethanol(EtOH,50mM),CSE(20–40μg/ml),orboth(CSE+EtOH),andanalyzedbyimmunoblotting,quantitativereversetranscriptionPCR,andcelldeathassays.SomecellswereincubatedwithMKC-3946,aninhibitorofendoplasmicreticulumtonucleussignaling1(ERN1,alsocalledIRE1)thatblocksXBP1sformation.MaleSprague-Dawleyratswerefedisocaloricamountsofanethanol-containing(Lieber-DeCarli)orcontroldietfor11weeksandexposedtocigarettesmokeorroomairinanexposurechamberfor2hourseachday.Duringthelast3weeks,asubsetofratsreceivedintravenousinjectionsoflipopolysaccharide(LPS,3mg/kgCorrespondence:AureliaLugea.Cedars-SinaiMedicalCenter;8700BeverlyBlvd,DavisBuildingD3096;LosAngeles,CA90048.Aurelia.Lugea@cshs.org.Authornamesinbolddesignatesharedco-firstauthorshipDisclosures:Theauthorsdisclosenoconflictsofinterest.Authorcontributions:AL,AGe,H-YS,CH,RTW,AGo,ZX,dataacquisitionandanalysis;SWF,electronmicroscopy;AL...
