[生物医药论文精品a]多糖修饰的巨噬细胞靶向超氧化物歧化酶及其体外降低辐射诱导的炎性细胞因子表达作用

A1A0A2A3A4A5A6A7A8A9A10A11A12A13A14A15A13A16A17A18A19A20A21A22A23A24A25A26A4A27A28A7A8A29A30A25A26A32A31A33A34A32A31A33A35A36A37A,BA38A39A40AA38A41A42A43A,BA38A44A45A46CA38A47A48A49CA38A41A50CA38A47A51A52A,BA53AA52A54A55A56A57A56A58A59A60A61A59A62A63A64A57A62A65A66A67A68BA52A54A55A56A69A70A71A72A73A63A64A65A66A74A75A68CA52A54A55A56A76A77A78A58A79A80A81A82A65A66A74A75A52A54A83A84250012A85TARGETEDDELIVERYOFSUPEROXIDEDISMUTASETOMACROPHAGESBYCHEMICALMODIFICATIONWITHPOLYSACCHARIDEANDATTENUATIONEFFECTSONRADIATIONINDUCEDINFLAMMATORYCYTOKINEEXPRESSIONINVITROJINFENGLIUA,B,LITENGA,CHUNHUILIUA,B,LIKUANHUC,YONGGANGWANGC,HONGLIUC,FENGSHANWANGA,BAINSTITUTEOFBIOCHEMICALANDBIOTECHNOLOGICALDRUG,SCHOOLOFPHARMACEUTICALSCIENCEBNATIONALGLYCOENGINEERINGRESEARCHCENTER,SHANDONGUNIVERSITYCCANCERCENTER,QILUHOSPITAL,SHANDONGUNIVERSITY,JINAN250012,CHINAFAX0086053188382548EMAILFSWANGSDUEDUCNABSTRACTTOIMPROVETHEABILITYOFSUPEROXIDEDISMUTASESODTOSUPPRESSREACTIVEOXYGENABBREVIATIONSSODCU,ZNSUPEROXIDEDISMUTASEROSREACTIVEOXYGENSPECIESTMCN,N,NTRIMETHYLCHITOSANCHLORIDETOWHOMCORRESPONDENCESHOULDBEADDRESSEDABBREVIATIONSSODCU,ZNSUPEROXIDEDISMUTASEROSREACTIVEOXYGENSPECIESTMCN,N,NTRIMETHYLCHITOSANCHLORIDETOWHOMCORRESPONDENCESHOULDBEADDRESSEDSPECIESROSMEDIATEDINJURY,CHEMICALLYMODIFIEDDERIVATIVESOFSODWITHN,N,NTRIMETHYLCHITOSANCHLORIDETMCANDHEPARIN,CATIONIZEDSODTMCSODANDANIONIZEDSODHEPARINSOD,WEREDESIGNEDANDPREPAREDINTHISSTUDY,THEINHIBITORYEFFECTOFTMCSODANDHEPARINSODONSUPEROXIDEANIONRELEASEFROMMACROPHAGESWASSTUDIEDINVITROBOTHTMCSODANDHEPARINSODEXHIBITEDEXCELLENTINHIBITORYEFFECTSONSUPEROXIDEANIONRELEASEFROMMACROPHAGES,ANDTHEEFFECTSOFTMCSODSURPASSEDTHOSEOFNATIVESODANDHEPARINSODTHEEFFECTSOFTMCSODANDHEPARINSODONINFLAMMATORYCYTOKINEEXPRESSIONINVITROWEREALSOEVALUATEDTHERESULTSSHOWEDTHATBOTHTMCSODANDHEPARINSODCOULDSIGNIFICANTLYLOWERTHELEVELSOFTRANSFORMINGGROWTHFACTOR1TGF1ANDINTERLEUKINE1IL1EXPRESSEDBYIRRADIATED3T3FIBROBLASTSTHESERESULTSDEMONSTRATEDTHATCATIONICPOLYSACCHARIDEORANIONICPOLYSACCHARIDESODDERIVATIVESMIGHTBEUSEFULINTHEPREVENTIONANDTREATMENTOFROSMEDIATEDINFLAMMATORYDISEASESTHISSTUDYALSODEMONSTRATEDTHATCHEMICALMODIFICATIONOFSOD,ESPECIALLYCATIONIZATION,GREATLYENHANCEDSODSINTRACELLULARDELIVERY,ANDASACONSEQUENCE,PRODUCEDASIGNIFICANTPROTECTIVEEFFECTAGAINSTROSMEDIATEDINJURYKEYWORDSSUPEROXIDEDISMUTASECHITOSANHEPARINCHEMICALMODIFICATIONCATIONICPOLYSACCHARIDEANIONICPOLYSACCHARIDE1INTRODUCTIONPHAGOCYTES,SUCHASMACROPHAGESANDNEUTROPHILS,SOMETIMESPRODUCEANEXCESSAMOUNTOFREACTIVEOXYGENSPECIESROSTHATCAUSEOXIDATIVEDAMAGE,ASOBSERVEDINVARIOUSINFLAMMATORYDISEASESTHESEFREERADICALSAREVERYUNSTABLEANDREACTRAPIDLYWITHOTHERSUBSTANCESORGROUPSINTHEBODY,LEADINGTOCELLORTISSUEINJURYTHISMAYBEILLUSTRATEDBYONEOFTHEMANYMECHANISMSWHICHOXIDATIVESTRESSCANCAUSEDAMAGEBYSTIMULATINGTHEFREERADICALCHAINREACTIONROSPRODUCEDBYMACROPHAGESPLAYACENTRALROLEINCAUSINGINFLAMMATORYDISEASES,SUCHASACUTELIVERFAILURE,ARTHRITIS,ANDSEPSIS13ROSACTIVATETRANSFORMINGGROWTHFACTOR1TGF1,ONEOFTHEMOSTIMPORTANTGROWTHFACTORSINTHEPATHOGENESISOFFIBROTICDISEASES,WHICHPROMOTESEPITHELIALCELLAPOPTOSIS4,5SUPEROXIDEDISMUTASESODISANENZYMETHATSCAVENGESROSANDHASTHEPOTENTIALTOTREATINFLAMMATORYDISEASESVIASUPPRESSINGROSPRODUCTIONBYMACROPHAGES68HOWEVERITSPOORHALFLIFEANDLOWAFFINITYTOMACROPHAGESHAVELIMITEDITSEFFECTIVENESSDIRECTMODIFICATIONOFSODISAPROMISINGAPPROACHTOIMPROVETHEDELIVERYPROPERTIESOFTHEENZYME9VARIOUSKINDSOFCU,ZNSODDERIVATIVESHAVEBEENDEVELOPEDVIACHEMICALMODIFICATIONFORTHETREATMENTOFLIVERORKIDNEYISCHEMIA/REPERFUSIONINJURIESINVIVO1012,INCLUDINGGLYCOSYLATEDANDPEGYLATEDSODSINOURLABORATORY,CU,ZNSUPEROXIDEDISMUTASECU,ZNSODHADBEENCHEMICALLYMODIFIEDWITHLOWMOLECULARWEIGHTHEPARINLMWH,ANDOURPREVIOUSSTUDYHADPROVEDTHATLMWHSODCONJUGATE,WHICHHADALOWERIMMUNOGENICITYANDHIGHERANTIINFLAMMATORYACTIVITY,ALSOGAINEDHIGHERSTABILITIESTHANCU,ZNSODTOWARDSACID,ALKALI,HEATANDTRYPSINTREATMENT10,13OUREARLYSTUDYALSOREVEALEDTHATHEPARINSODCOULDATTENUATEBLEOMYCINBLMINDUCEDLUNGFIBROSIS14INTHISSTUDYHEPARINSODWASALSOPREPAREDUSINGTHESAMESTRATEGYANOTHERSODCHEMICALMODIFICATIONSTRATEGYISDIRECTCATIONIZATIONCATIONIZATIONOFPROTEINSISKNOWNTOIMPROVETHEIRCELLULARUPTAKEVIANONSPECIFICELECTROSTATICINTERACTIONSWITHTHENEGATIVECHARGEOFTHECELLSURFACE,ASHASBEENDEMONSTRATEDWITHSOD1518MITSURUETALHADREPORTEDTHATDIRECTCATIONIZATIONOFSODGREATLYENHANCEDITSCELLULARUPTAKEANDSUBSEQUENTLYIMPROVEDITSINHIBITORYEFFECTONSUPEROXIDEANIONRELEASEFROMMACROPHAGES19CHITOSANISAPOSITIVELYCHARGEDPOLYSACCHARIDECOMPOSEDOF2AMINO2DEOXY14DGLUCOPYRANOSE,WHICHHASREACTIVEAMINOANDHYDROXYLGROUPSCHITOSANHASATTRACTEDMUCHATTENTIONASABIOMEDICALMATERIAL,OWINGTOITSUNIQUEBIOLOGICALACTIVITIES,SUCHASANTITUMOR,ANTIBACTERIAL,ANDIMMUNOSTIMULATORYACTIVITIES20RECENTLY,THEANTIOXIDANTACTIVITYANDRADIOPROTECTIVEEFFECTOFCHITOSANALSOSTIMULATESCIENTISTSINTERESTS21,22ITWASPROVEDTHATLOWMOLECULARWEIGHTCHITOSANHADAHIGHRADICALSCAVENGINGACTIVITYFOROHANDLOWRADICALSCAVENGINGABILITYFOR2O23,24N,N,NTRIMETHYLCHITOSANCHLORIDETMC,PROTONATEDCHITOSAN,POSSESMOREPOSITIVECHARGETHANCHITOSANWESPECULATETHATTMCSODPOSSIBLYHASHIGHAFFINITYTOSOMECELLS,SUCHASMACRO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HIOCYANATEFITCBYTHEMETHODOFMONSIGNYETAL15MACROPHAGES1106CELLS/WELLCULTUREDIN12WELLPLASTICPLATEWEREINCUBATEDWITH1MLOFSERUMFREECULTUREMEDIUMCONTAININGFITCLABELEDNATIVESODORSODDERIVATIVES200GPROTEIN/MLAT37CAFTERAPROPERTIME,THECELLSWEREWASHEDTHREETIMESWITHICECOLDPBSANDANALYZEDBYFLOWCYTOMETRYFOREACHSAMPLE,10,000CELLSWEREANALYZEDBYTHELOGARITHMICAMPLIFICATIONOFFLUORESCENCEINTENSITYFI24EXPERIMENTOFTHEEFFECTSOFSODCONJUGATESONRADIATIONINDUCEDINFLAMMATORYCYTOKINEEXPRESSIONINVITRO241CELLCULTUREMOUSELUNGFIBROBLASTCELLLINE,3T3,WASOFFEREDBYSHANDONGACADEMYOFMEDICALSCIENCECHINATHECELLSWERECULTUREDINRPMI1640MEDIUMSUPPLEMENTEDWITH10FBSCONTAINING100U/MLOFPENICILLINGANDSTREPTOMYCINCELLSWEREPASSAGEDBYTRYPSINTREATMENTANDINCUBATEDUNDERANATMOSPHEREOF5CO2/95AIRAT37CCELLVIABILITYWASMORETHAN95ASMEASUREDBYTRYPANBLUEDYEEXCLUSION242GROUPINGANDRADIATIONADMINISTERINGCELLSWERETRYPSINIZEDANDSEEDEDONTOA48WELLPLATEATADENSITYOF2104CELLS/WELL,ANDDIVIDEDINTO12GROUPSINORMALCONTROLGROUPNORADIATIONADMINISTEREDANDTREATMENTIICONTROLGROUPPBSIIITMCGROUPTMCCONCENTRATIONWASIDENTICALTOTHEPORTIONINTMCSODIVHEPARINGROUPHEPARINCONCENTRATIONWASIDENTICALTOTHEPORTIONINHEPARINSODVTMCSODGROUPTMCANDCU,ZNSODMIXTURE,SODCONCENTRATIONIDENTICALTOTHATOFSODGROUPANDTMCCONCENTRATIONIDENTICALTOTHATOFTMCGROUPVIHEPARINSODGROUPHEPARINANDCU,ZNSODMIXTURE,WITHSODCONCENTRATIONIDENTICALTOTHATOFSODGROUPANDHEPARINCONCENTRATIONIDENTICALTOTHATOFHEPARINGROUPVIICU,ZNSODADDEDPREIRRADIATIONGROUPVIIIHEPARINSODADDEDPREIRRADIATIONGROUPTMCA89SODADDEDPREIRRADIATIONGROUPA90SODADDEDPOSTIRRADIATIONGROUPA91TMCSODADDEDPOSTIRRADIATIONGROUPA92HEPARINSODADDEDPOSTIRRADIATIONGROUPTHEFINALENZYMEACTIVITIESINALLGROUPSRELATINGTOSODORITSDERIVATIVESWEREIDENTICAL,250U/MLTHREEPARALLELWELLSWEREINCLUDEDINEACHGROUPAFTER24HINCUBATION,THECELLCULTUREWASEXPOSEDTO12GYOF6MVXRADIATIONTHETESTEDSAMPLESOFGROUPA93A89WEREADDED2HBEFOREIRRADIATION,WHILETHETESTEDSAMPLESOFGROUPA90TOA92WEREADDEDIMMEDIATELYAFTERIRRADIATION243DETERMINATIONOFCYTOKINETGF1ANDIL1THEDETERMINATIONSOFTGF1ANDIL1WEREDONEACCORDINGTOTHETESTKITINSTRUCTIONS244DATAANALYSISTHEDATAWASANALYZEDBYUSINGSPSS110STATISTICALSOFTWARESTATISTICALCOMPARISONSBETWEENTHEGROUPSWERECARRIEDOUTWITHONEWAYANOVAWITHFISHERSEXACTTEST,P005COMPARINGTMCSODORHEPARINSODADDEDPOSTIRRADIATIONWITHPREIRRADIATIONGROUP,ITCOULDBEFOUNDTHATTGF1CONTENTOFADDEDPREIRRADIATIONGROUPWASLOWERP005THERESULTSALSOSHOWEDTHATTHEEFFECTOFSODANDSODDERIVATIVESONRADIATIONINDUCEDIL1WASVERYSIMILARWITHTHATOFTGF1FIG5B4DISCUSSIONSODCANBEUSEDASATHERAPEUTICAGENTSINCETHISENZYMECANELIMINATETHESUPEROXIDEANIONS,WHICHPLAYANIMPORTANTROLEINVARIOUSDISEASESMEDIATEDBYROS,SUCHASINFLAMMATIONHOWEVER,THEMAJORDRAWBACKOFSYSTEMICFIG5A,BFIG4ADMINISTRATIONOFANTIOXIDANTENZYMESLIKESODISTHEIRULTRASHORTELIMINATIONHALFLIFEANDTHEIRLOWAFFINITYTOCELLMEMBRANESALSO,THEDAMAGEINDUCEDBYROSISCHARACTERIZEDBYSITESPECIFICEVENTSTHEREFORE,IFANTIOXIDANTENZYMESARETOBEUSEDFORTHETREATMENT,THEYSHOULDBELOCATEDINCLOSEPROXIMITYTOTHESITEOFINFLAMMATIONITISPROVEDTHATCHEMICALMODIFICATIONOFSODISAPROMISINGWAYTOCONTROLITSDISPOSITIONCHARACTERISTICSINTHEBODYANDCONSEQUENTLYIMPROVEITSPHARMACOLOGICALACTIVITY14,28,29DIRECTCHEMICALMODIFICATIONOFSODINCLUDEDCATIONIZATION,GLYCOSYLATION,SUCCINYLATIONANDPEGALATIONCATIONIZATIONISAUNIVERSALAPPROACHTHATCANBEAPPLIEDTOINCREASETHEINTERACTIONOFCOMPOUNDSWITHNEGATIVELYCHARGEDBIOLOGICALCOMPONENTS30PROTEOGLYCANSANDOTHERANIONICMOLECULESHAVEBEENSHOWNTOBETHESITESONTHECELLMEMBRANETHATINTERACTWITHCATIONICCOMPOUNDSTHROUGHELECTROSTATICINTERACTIONS18,31SUCHELECTROSTATICINTERACTIONSMAYINCREASETHEDELIVERYOFTHERAPEUTICCOMPOUNDSTOTHETARGETCELLSCATIONIZATIONOFPROTEINSASATOOLTOINCREASETHEIRRESIDENCETIMEINTARGETORGANSHASBEENSUGGESTEDINTHEPAST18CATIONIZATIONANDGLYCOSYLATIONHAVEALSOBEENAPPLIEDTOSODANDEFFECTSINVOLVINGINTERACTIONWITHTISSUESANDBIOLOGICALACTIVITIESHAVEBEENREPORTED13,14,19HOWEVER,LITTLEATTENTIONHASBEENPAIDTOCHITOSAN,ACATIONICPOLYSACCHARIDE,USEDASAPROTEINMODIFICATIONMATERIALTHEREFORE,INTHEPRESENTSTUDY,TWOPOLYSACCHARIDES,CHITOSANANDHEPARIN,WERESELECTEDTOOBTAINCATIONIZEDSODANDANIONIZEDSODROSWEREMAINLYPRODUCEDBYSURFACENEGATIVELYCHARGEDMACROPHAGESSINCESODISALSONEGATIVELYCHARGED,ONEPOSSIBLEWAYFORSODTOATTACHTOMACROPHAGESISTOCHANGEITSELECTRICCHARGEPROPERTYTHEABILITYOFCATIONIZEDPROTEINSTOADHERETONEGATIVELYCHARGEDTISSUESPOSESANINTERESTINGPOTENTIALINDRUGTARGETINGDELIVERYINTHISSTUDY,WEHYPOTHESIZEDTHATTHEATTACHMENTOFANTIOXIDANTENZYMESTOTHESITEOFINFLAMMATIONWOULDINCREASETHEEFFICACYOFINFLAMMATIONTHERAPYWITHTHEENZYMESCHITOSANISALINEARHOMOPOLYSACCHARIDECONSISTINGOFDGLUCOSAMINEMONOMERSLINKEDTHROUGH14GLYCOSIDICLINKAGESITISACATIONICPOLYSACCHARIDEMADEFROMALKALINENDEACETYLATIONOFCHITINASANATURALPRODUCT,ANTIOXIDANTACTIVITYOFCHITOSANANDITSDERIVATIVESHAVEATTRACTEDMUCHATTENTION3234JEONETAL35FOUNDTHATCHITOSANHADANANTIOXIDANTEFFECTONCHRONICCCL4INDUCEDHEPATICINJURYTHEYALSOFOUNDTHATTHEANTIOXIDANTENZYMECATALASEANDSODACTIVITIESINLIVERTISSUEWEREINCREASEDBYCHITOSANTMC,ADERIVATIVEOFCHITOSANWITHHIGHPOSITIVECHARGE,ISSOLUBLEINTHEENTIREPHRANGEANDHASBEENPROVENTOBEAPOTENTABSORPTIONENHANCEROFPEPTIDEDRU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