Biological Effects of Cigarette Smoke in Cultured Human Retinal Pigment Epithelial Cells 英文参考文献.docVIP

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Biological Effects of Cigarette Smoke in Cultured Human Retinal Pigment Epithelial Cells 英文参考文献.doc

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Biological Effects of Cigarette Smoke in Cultured Human Retinal Pigment Epithelial Cells 英文参考文献

BiologicalEffectsofCigaretteSmokeinCulturedHuman RetinalPigmentEpithelialCells AliceL.Yu1*,KerstinBirke2,JohannesBurger1,UlrichWelge-Lussen2 1Department of Ophthalmology, Ludwig-Maximilians-University, Muenchen, Germany, 2Department of Ophthalmology, Friedrich-Alexander-University, Erlangen, Germany Abstract Thegoalof thepresentstudy wastodeterminewhethertreatmentwithcigarette smokeextract (CSE)inducescellloss, cellularsenescence,andextracellularmatrix(ECM)synthesisinprimaryhumanretinalpigmentepithelial(RPE)cells.Primary culturedhumanRPEcellswereexposedto2,4,8,and12%ofCSEconcentrationfor24hours.Celllosswasdetectedbycell viability assay. Lipid peroxidation was assessed by loss of cis-parinaric acid (PNA) fluorescence. Senescence-associated ?- galactosidase(SA-?-Gal)activitywasdetectedbyhistochemicalstaining.ExpressionofapolipoproteinJ(ApoJ),connective tissuegrowthfactor(CTGF),fibronectin,andlamininwereexaminedbyreal-timePCR,westernblot,orELISAexperiments. Theresultsshowedthatexposureofcellsto12%ofCSEconcentrationinducedcelldeath,whiletreatmentofcellswith2,4, and8%CSEincreasedlipidperoxidation.Exposureto8%ofCSEmarkedlyincreasedthenumberofSA-?-Galpositivecellsto upto82%,andthemRNAexpressionofApoJ,CTGF,andfibronectinbyapproximately3–4fold.Treatmentwith8%ofCSE alsoincreasedtheproteinexpressionofApoJandCTGFandthesecretionoffibronectinandlaminin.Thus,treatmentwith CSE can induce cell loss, senescent changes, and ECM synthesis in primary human RPE cells. It may be speculated that cigarettesmokecouldbeinvolvedincellulareventsinRPEcellsasseeninage-relatedmaculardegeneration. Citation:YuAL,BirkeK,BurgerJ,Welge-LussenU(2012)BiologicalEffectsofCigaretteSmokeinCulturedHumanRetinalPigmentEpithelialCells.PLoSONE7(11): e48501.doi:10.1371/journal.pone.0048501 Editor:MichaelE.Boulton,UniversityofFlorida,UnitedStatesofAmerica ReceivedJune21,2012;AcceptedSeptember26,2012;PublishedNovember14,2012 Copyright:?2012Yuetal.Thisisanopen-accessarticledistributedunderthetermsoftheCreative