in utero sflt-1 exposure differentially affects gene expression patterns in fetal liver v. stojanovska参考-匠人.pdf

该【in utero sflt-1 exposure differentially affects gene expression patterns in fetal liver v. stojanovska参考-匠人 】是由【妙玉】上传分享，文档一共【9】页，该文档可以免费在线阅读，需要了解更多关于【in utero sflt-1 exposure differentially affects gene expression patterns in fetal liver v. stojanovska参考-匠人 】的内容，可以使用淘豆网的站内搜索功能，选择自己适合的文档，以下文字是截取该文章内的部分文字，如需要获得完整电子版，请下载此文档到您的设备，方便您编辑和打印。JournalofDevelopmentalInuterosFlt-1exposuredifferentiallyaffectsOriginsofHealthandDiseasegeneexpressionpatternsinfetalliver/,,,-Schakel1,,,?sch11DepartmentofObstetricsandGynecology,UniversityofGroningen,UniversityMedicalCenterGroningen,Theherlands,2DepartmentofPathologyandMedicalBiology,UniversityofGroningen,UniversityMedicalCenterGroningen,herlands,3DepartmentofPathologyandMedicalBiology,DivisionofMedicalCitethisarticle:StojanovskaV,HolwerdaBiology,UniversityofGroningen,UniversityMedicalCenterGroningen,herlandsand4Nutrition,KM,vanderGraafAM,VerkaikSchakelRN,MetabolismandGenomicsGroup,DivisionofHumanNutrition,WageningenUniversity,herlandsBoekschotenMV,FaasMM,ScherjonSA,Pl?schT(2019)InuterosFlt--liketyrosinekinasefactor1(sFlt-1):,littleisknownabouttheeffectsofsFlt--1concentrationduringReceived:-DawleyRevised:18September2018dams(sevenfemales,10weeksold)duringmid-gestation(gestationalday8)withadenovirusAccepted:26September2018overexpressingsFlt-1,andage-matchedcontrols(sixfemales,10weeksold)withemptyKeywords:adenoviralvirusinordertoquantifythesFlt-;developmentalstage;ics;exposedtoadenoviralsFlt-1deliveryweresubdividedintoalow(n=4)andhighsFlt-1fetus;general;molecular/cellular;small(n=3)-wayanalysisofvarianceshowedthatanimalsfetuses(fiveperdam)exposedtohighsFlt-1concentrationsinuteroshowfetalgrowth±±±<Addressforcorrespondence:restriction(-;mean(M).;),TorstenPl?sch,,themicroarrayanalysisofGynecology,UniversityofGroningen,thefetalliverofthehighsFlt-1groupshowedsignificantenrichmentofkeygenesforfattyUniversityMedicalCenterGroningen,PB3000,,usingpyrosequencing,wefoundthatthe9700RBGroningen,-mail:t.******@-paniedbydecreasedmethylationofitspromoter(±%methylationinhighsFlt-±%methylationincontrol,M±.,P<).OurdatashowthathighsFlt--,toxins,smokingandmaternaldiseasesduringpregnancyaffectthebirthweightoftheoffspring,resultinginan–,theoppositesituationofincreasedbirthweightormacrosomiaalsocontributestometabolic–programmingandlater-lifemorbidity,-associateddisorder,plicatedwithfetalgrowthrestrictionandaffects3–7%,itisassociatedwithseveralpathophysiologicalpathwaysincludingangiogenicimbalancewithapredominanceofsolublefms-liketyrosinekinase-1(sFlt-1),whichincreasedtwo-tofive-,11sFlt-1servesasanantagonisttoplacentalgrowthfactor(PlGF)andvascularendothelialgrowthfactor(VEGF),thuspromotinghypovascularization,--1showsincreasedapolipoproteinfractionsthataremediatorsofcholesterolesterificationandremovalofexcesscholesterolfrom?,thefattyacidavailabilityisincreasedinsFlt-1-inducedpreeclampsiadueInternationalSocietyforDevelopmentaltoincreasedexpressionoffattyacidtranslocaseinsFlt-,itwasreportedthattheliverXreceptorandretinoidXreceptor(LXR/RXR)pathwayisupregulatedinsFlt-1--1contributetothepathogenesisofpreeclampsiaandfetalgrowthrestriction,15thespecificeffectsofsFlt-1onthedevelopingfetusarenotwellDownloadedfrom,on11Apr2019at16:33:22,subjecttotheCambridgeCoretermsofuse,availableathttps://e/:///.,weaimedtoevaluatethefollowing:,bloodwas(1)theeffectsofsFlt-1onthefetaldevelopment,(2)effectsoncollectedviatheabdominalaortaandcentrifugedasstatedabovefetallivergeneexpressionand(3),,anditwascollectedinheparin-coatedcapillarytubes(VitrexMedicalA/S,Herlev,Denmark).sFlt-1concentrationsMaterialsandmethodsinplasmaweredeterminedusingamousesFlt-1enzyme-linkedAnimalsimmunosorbentassaykit(R&DSystems,Inc.,Minneapolis,MN,USA).Atotalof24urinesampleswerecollectedbyplacingagesatGD16and17,(mg)-Dawley(SD)ratsof10-weeks-old24-hurinevolume.(CharlesRiver,France)werehousedinalightandtemperature-controlledfacility(lightsonfrom7:00amto7:00pm,21°C).-estrusphaseoftheirTotalRNAwasisolatedfromfetalliverscollectedonGD19withcycle,,allprepDNA/RNAminikit(Qiagen,Venlo,herlands),ifspermatozoawerepresentinthevaginalsmear,thenitwasfollowingthemanufacturer’,animalswerewereassessedwithNanodrop2000c(ThermoScientific,Pitts-=randomlydividedintotwogroups:control(n6)andsFlt-1burgh,PA,USA).RNAwasimmediatelystoredat?80°Cuntil=(n7),receivingeithercontrolAd-nulloradenovirusover--1(Ad-sFlt-1),,usingtheEukaryotetotalRNAnanoassay><eitherultra-high(1000ng/ml)orverylow(1000ng/ml)sFlt-ordingtomanufacturer’sprotocol(AgilentTechnologies,plasmaconcentrationsinoursFlt-1group,andwecontinuedtheAmsterdam,herlands).SamplesthathadanRNAintegrity==analysiswithhigh(n3)andlowsFlt-1(n4)>-1andcontroladenovirusMicroarrayhybridizationandanalysisAdenovirusvectorstockofAd-null(,UniversityMedicalCenterGroningen,herlands)TotalRNAisolatedfromfetallivers(fivesamplespergroup:andAd-sFlt-1(,BethIsraelcontrolandhighsFlt-1),Boston,MA,USA)(LifeTechnologies,,CA,USA)andAffymetrixGeneChipWTTerminalwasperformedwithacesiumchloride(CsCl)densitygradientLabelingKit(Affymetrix,SantaClara,CA,USA).Allsamples(d=).(Biorad,Temse,Belgium)-formingunits(PFU)wasandnormalizationwasperformedwithBioconductorsoftwareanalyzedwithanenzyme-linkedimmunoassaythatdetectsthepackages(Lin2011).Probessetswereassignedtouniquegeneadenoviralhexon(Adeasyviraltiterkit,AgilentTechnologies,identifiers(IDs)oftheEntrezGenedatabase,,CA,USA),and1×1012PFUofadenovirusexpres--1ortheemptyvectorwereinjectedviathetailveinongestationalday(GD),andonlytheoneswithanintensityvalueof>20onatleastthreearraysandinterquartileOnGD19,thesystemicbloodpressureforallgroupswasassessedrange(IQR)>(100%O2,,5%%)ponentanalysis(PCA)usingtheabdominalaortaandconnectedwithabed-,which(Datex-Ohmeda,Cardiocap/5).Bloodpressuremeasurementrepresentfoldchanges,-1groupusingintensity--,e-analysis(GSEA)-?parisonofgeneexpressionpatternswasdonewithplasmawasstoredat80°,://,on11Apr2019at16:33:22,subjecttotheCambridgeCoretermsofuse,availableathttps://e/:///-,somedamsshowedamoderateincrease(n=4)ofsFlt-1concentrationsinplasma(sFlt-1atGD12:OftotalRNA,1μgwasusedforcDNAconversionwithreagents310±109ng/mlandatGD19:640±624ng/ml),whereasotherfromInvitrogen(Invitrogen,USA),accordingtomanufacturer’sdams(n=3)showedhighsFlt-1concentrations(atGD12:-timequantitativepolymerasechain1073±735ng/ml;andatGD19:4726±2215ng/ml).Inthatreaction(qPCR)wasperformedusingPowerUpSYBRgreenway,wecharacterizedtwogroupsofdamswithlowandhighmastermix(AppliedBiosystems,FosterCity,CA,USA)onSte-sFlt-1concentrationsinadditiontothecontrolgroup,wherethepOnePlusreal-timePCRsystem(AppliedBiosystems)withthesFlt-1concentrationsatGD19werealmostzerofollowingsettings:20s95°C,40cyclesat95°Cfor1sandat60°C(±).(,astheprimarypurposewasto)weredesignedwithPrimerExpressSoftwareshowincreaseofsFlt-1concentrationsaftersFlt-1adenoviral(AppliedBiosystems)andsynthesizedbyEurogentec(Seraing,,atGD19theaorticbloodpressureBelgium).Theaverageexpressionlevelof36b4andGapdhwaswasassessed,andtherewerenosignificantdifferencesbetweenusedasahousekeepinggeneinallqPCRanalysisandtheddCtthecontrolandlowsFlt-,thehighsFlt-().Furthermore,excessiveproteinuriawasnotpresentinanyofGenomicDNAisolationandpyrosequencingtheinspectedgroups().FetalliverswerehomogenizedwithaTissueLyserLT(Qiagen)-1concentrationsare’RNAminikit(Qiagen),,while500ngofgenomicDNAwasusedforbisulfiteconversionwithIthasbeenreportedthatincreasedsFlt-1concentrationsinthe15theEZDNAmethylationgoldki