Manyreportshavepointedoutthatultraviolet(UV )caninducetheagingofskinanderythemat ic<1,2 > .TheoxidativedamageofUVonskinaswellasoncellisbyincreasingtheproductionoffreeradi cals,whichcoulddamageDNA ,proteinandunsatu ratedfattyacidinthecellmembrane<3,4 > .SomereportsindicatedthatnaturalantioxidantscouldinhibittheoxidativeinjurescausedbyUV.Mostreportsmentionedthatthenaturalantioxidantsmainlycomefrom plantsorherbs ,whichhavebeenusedinmedicalandhealthcare ,butfewofthemfrommarineproducts .PCFisabioactivepolypeptidefrommarineChlamysFarreriandhastheantioxida tiveroleinsomepreliminarytest<5> .Inthisstudyweestablishedanoxidative damagemodelofHelacellsandstudiedtheprotectiv eeffectofPCFonthecellsanditspossiblemechanism.1 MaterialsandMethods1 .1 DrugsandreagentsPolypeptidesisolatedfromC hlamysFarreri(PCF)waspurifiedandanalyzedbyHPLC,storedat4℃andisolatedbyusingbiologicalengineeringtech nique .HelacellswereboughtfromShandongMedicalScienceInstitute .Malondialdehyde (MDA)testkitsandenzymetestkitsincludingglutathioneperoxidase(GSH px) ,superoxidedismutase (SOD)andcatalase(CAT)boughtfromNanjingJianchengBioengineer ingInstitute .1 .2 Methods1 .2 .1 TheestablishmentofoxidativemodelforHelacellsdamagedbyUV SuspensionofHelaep itheliacells (5× 1 0 8/L)culturedfor 2 4hourswerepreparedandput 1mLofthesuspensionintoeachholeof 2 4 holeplates .SomeHelacellswererandomlydividedintosixgroups :thecontrol (C) group ,modal(M ) group ,5 g/L PCF group ,1 0 g/L PCF group ,2 0 g/LPCFgroupand 1 0 g/LVita min C (Vc ) group .AfterincubationwithRPMI 16 40 ,PCF ,Vc ,respectively ,for 1 0minutes ,thecellswereexposedtoUVA (irradiationintensity :3.6 5 0mJ/cm2 ) ,excepttheCgroup .Theintracellularfreecalcium (Ca2 + ) ,apoptosisandcelldeathratewerethenmeasuredbyflowcytometry (FCM ) .En zymeactivitiesandMDAweredeterminedbybio chemicalassays .TheothermiceweretreatedalmostthesameasmentionedabovebuttheywereirradiatedbyUVB (irradiationintensity :7.1 5× 1 0 - 5J/cm2 ) .1 .2 .2 Thedetectionofapoptosisandcelldeathrate Thecellswereculturedcontinuouslyat37℃ ,φ =0 .0 5CO2 fortwohours,thenthecellswerecollectedandstainedwithAnnexinVandPI .TheapoptosisandcelldeathrateweremeasuredbyFCM .1 .2 .3 ThedetectionofintracellularfreecalciumThecellswereculturedcontinuouslyfor 2hours ,theywerethencollectedandstainedwithFluo 3AM .Then ,theintracellularfreecalcium (Ca2 + )wasmeasuredbyFCM .1 .2 .4 Thedetectionoftheactivitiesofsuccinatedeh ydrogenase TheMTTwereaddedintothecellsuspension.Aftercontinuousculturefor 4hours ,thevaluesofsuccinatedehydrogenase (SDH )inthecellsweremeasuredbyenzymelabeledchromometerat 4 90nm .1 .2 .5 ThedetectionoftheactivitiesofGSH px ,SOD ,CATandthecontentsofMDA TheactivitiesofGSH px ,SOD ,CATandthecontentsofMDAinthesupernatantsofth ecellswereseparatelydetectedbybiochemicalmethods.2 Results2 .1 TheeffectsofPCFontheapoptosisandcelldeath ofHelaepitheliacellsirradiatedbyUVAandUVBTh eapoptosisandcelldeathofHelacellsinMgroupsignificantlyincreasedthanthoseinCgroup(F =5 8.6 1 - 6 7.2 2 ,q =7.85 - 1 8.5 6 ,P <0 .0 1 ) .TheapoptosisandcelldeathofHelacellsinPCFgroupwerelowerthanthatofM group (q =3.77-8.93,P <0 .0 5 ) (Table 1 ) .TheapoptosisandcelldeathrateinHelacellsin 2 0 g/Lgroupwerelowerthanthatof 5 g/Lgroupand1 0 g/Lgroup (q =1 .6 6 - 7.95 ,P <0 .0 5 ) .Table 1 TheeffectsofPCFonapoptosisandcelldeathrateofHelacellsdamagedbyUVAandUVB (χ % , x±s)GroupnUVAUVBapoptosisrate celldeathrateapoptosisrate celldeathrateC 3 0 .55± 0 .0 2 0 .96± 0 .0 1 0 .55± 0 .0 2 0 .96± 0 .0 1M 3 3 .3 5± 0 .0 2 3 .45± 0 .0 2 3 .64± 0 .0 2 4.1 0± 0 .0 2 5g/LPCF 3 3 .1 6± 0 .0 3
△ 3 .41± 0 .0 1 △ 3 .1 3± 0 .0 1 △ 3 .3 2± 0 .0 4△1 0 g/LPCF 3 3 .0 5± 0 .0 5△ 3 .3 5± 0 .0 3 △ 3 .0 9± 0 .0 5△ 3 .2 1± 0 .0 3 △2 0 g/LPCF 3 2 .0 3± 0 .0 1 △ # 1 .75± 0 .0 1 △ # 1 .63± 0 .0 2 △ # 1 .47± 0 .0 7△ # MvsC ,F =58.61 -67.2 2 , q =7.85-1 8.56,P <0 .0 1 ;PCFvsM ,△ q =3 .77-8.93 ,P <0 .0 5;2 0g/LPCFvs 5g/Land 1 0g/LPCF ,# q =1 .66-7.95,P <0 .0 52 .2 TheeffectsofPCFontheintracellularCa2 +andactivitiesofSDHinHelaepitheliace
More abstracts about the 扇贝多肽对紫外线损伤Hela细胞的保护作用(英文)