phosphodiesterase 5 inhibition ameliorates angiotensin ii-dependent hypertension and renal vascular dysfunction manuel thieme资料.pdf

该【phosphodiesterase 5 inhibition ameliorates angiotensin ii-dependent hypertension and renal vascular dysfunction manuel thieme资料 】是由【彩屏】上传分享，文档一共【8】页，该文档可以免费在线阅读，需要了解更多关于【phosphodiesterase 5 inhibition ameliorates angiotensin ii-dependent hypertension and renal vascular dysfunction manuel thieme资料 】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。AmJPhysiolRenalPhysiol312:F474–F481,,2017;doi:.-dependenthypertensionandrenalvasculardysfunctionManuelThieme,1*,1*EvanthiaMergia,,1GuangYang,1LydiaHering,1KatharinaGrave,1HenningHoch,,1andJohannesStegbauer11DepartmentofNephrology,MedicalFaculty,UniversityHospitalDüsseldorf,Heinrich-Heine-UniversityDüsseldorf,Düsseldorf,Germany;and2DepartmentofPharmacologyandToxicology,Ruhr-UniversityBochum,Bochum,GermanySubmitted29June2016;acceptedin?nalform3January2017DownloadedfromThiemeM,SivritasSH,MergiaE,PotthoffSA,YangG,Heringvasocontractilityandimpairedvasorelaxation,resultinvascu-L,GraveK,HochH,RumpLC,,leadingtohypertension(8).inhibitionamelioratesangiotensinII-dependenthypertensionandre-Therenin-angiotensinsystem(RAS)andtheNO/:F474–signalingcascadearekeyplayersintheregulationofvascularF481,,2017;doi:.Inthisregard,angiotensin(Ang).—Changesinrenalhemodynamicshaveamajorimpactinjurybyinducingvasoconstriction,vascularsmoothmusclehttp://ajprenal./onbloodpressure(BP).Angiotensin(Ang)IIhasbeenshowntocellproliferationandhypertrophy,andvascularin?ammationinducevasculardysfunctionbyinteractingwithphosphodiesteraseaswellasextracellularmatrixdegradation(21,33).Further-(PDE),wemore,angiotensinII(AngII)infusionhasbeenshowntomeasuredtheeffectsofPDE5(sildena?l)orPDE1(vinpocetine)decreaseNObioavailabilty,leadingtovasculardysfunction(2,inhibitiononrenalblood?ow(RBF),BP,andrenalvascularfunction12,14,15,34).BesidestheimpactofAngIIonNObioavail--termAngability,severalstudieshavehighlightedtheroleofAngIIinIIinfusion,sildena?ldecreasedBPandincreasedRBFinC57BL/6modulatingcGMPdegradationbyin?uencingtheactivityof(WT),-degradingphosphodiesterases(PDE)invascularAdditionally,renalcGMPlevelsweresigni??lbutnotaftervinpocetineinfusion,indicatingapre-expressionofPDE1andPDE5,,,2017AngIIinfusion(500ng·kg?1·min?1)increasedBPandledtoim-(10,16,17).Ingeneral,PDE1isactivatedbyincreasedpairedNO-???intracellularCaconcentrations,whichareobservedduringtreatmentwithsildena?l(100mg·kg1·day1)attenuatedAngII-AngIImediatedvasoconstriction(13,30,31).PDE5isacti-dependenthypertensionandimprovedNO-,urinarynitriteexcretion,amarkerfordomainofPDE5(29).Thus,PDE5limitscGMPinducedrenalNOgeneration,wasincreasedinWTmice,whereasrenalcGMPvasorelaxationthroughacGMP-mediatednegativefeedbackgenerationwasdecreasedandrestoredaftersildena?ltreatment,,therehasbeencon?,thepredominantPDEregulatingvascularfunctioninresistancewenextanalyzedeNOS-KOmice,amousemodelcharacterizedbyarteriesduringAngII--KOmice,chronicAngII(10)reportedthatchronicAngIIinfusionreducescGMPinfusionincreasedBPbutdidnotimpairNO---Moreover,sildena?ldidnotin?uenceBPorvascularfunctionintrast,weandothershaveshownthatvasculardysfunctionineNOS--,whichresultedinanexaggeratedPDE5activationand?nallyinreducedNO/cGMPmediatedphosphodiesterase;renalblood?ow;angiotensin;cGMP;hyperten-relaxation(34).,weexaminedwhetheracuteandchronicinhibitonHYPERTENSIONISTHELEADINGRISKFACTORforcardiovascularofaspeci?-,,-siblefederalstateauthority(LandesamtfuerNatur-,Umwelt-,und*-Westfalen;reference:-Addressforreprintrequestsandothercorrespondence:,)ordingtotheguidelinesfromofNephrology,MedicalFaculty,üsseldorfHeinrich-Heine-Directive2010/63/EUoftheEuropeanParliamentontheprotectionofUniversityDüsseldorfMoorenstrasse5,40225Düsseldorf,Germany(e-mail:animalsusedforscienti?-to14-week-oldmalejohannes.******@-).C57BL/6Jwild-type(WT)miceandendothelialnitricoxidesynthase-F4741931-857X/17Copyright?2017theAmericanPhysiologicalSocietyPDE5AFFECTSHYPERTENSIONANDRENALVASCULARDYSFUNCTIONF475knockout(eNOS-KO)mice(?decke,dose-responsecurvestoGSNOwererecordedinthepresenceorCardiovascularPhysiology,Dusseldorf,Germany)werebredandabsenceofeithersildena?l(300nM)orvinpocetine(1?M).IsometrichousedinthelocalanimalcarefacilityoftheUniversityHospitalforcesareexpressedasapercentageofthemaximalresponsetoDüsseldorf,asdescribedpreviously(11).?--KOmiceAcutepressorresponsesandrenalblood?owtovinpocetineandwereinfusedcontinuouslywithAngII(500ng·kg?1·min?1;Sigma-sildena?lweremeasuredinanesthetizedmic,easdescribedpreviouslyAldrich)viaosmoticminipumps(model1002;Alzet,Cupertino,CA)(32).Inbrief,(100mg/kg)andxylazine(5mg/kg)anddecapitatedattheendofthe[intraperitonealanesthesiawithketamine(100mg/kg)andxylazine(-mg/kg)].FourdaysbeforeAngIIinfusion,WTandeNOS-?uids,AngII,andPDEweredividedintoasildena?l(100mg·kg?1·day?1)andvehicle-?lwasdissolvedintapwateracidi?edtopH3measuringrenalblood?ow(RBF),asmallincisionwasmadeontheataconcentrationof400mg/l,resultingintheingestionof~100left?anktoexposethekidney,andanultrasonic?owmeterinterfacedmg·kg?1·day?1andinsuf?cientsildena?lplasmalevels(1,34).Thewitha5-mmV-shapedprobewasplacedaroundtheleftrenalarteryvehiclegroupreceivedonlytapwateracidi?edtopH3.(;TransonicSystems,Ithaca,NY).(BP)wasDownloadedfromMicewereallowedtostabilizefor30minbeforemeasurementsweremeasuredinconsciousmicebytail-cuffplethysmography(BP-98A;).Forhabituation,beforetheexperiment,,,10measurementspermousePDE5inhibitorsildena?lorthePDE1inhibitorvinpocetinewaswererecordeddaily,startingonthedaywhenthemicewereassignedadministeredinincreasingdoses(,,,?lorvehicle-)at5-(iplast,Gams,Swit-hypertensiveconditions,AngII(200ng·min·kg)wasappliedhttp://ajprenal./zerland)wereutilized,--hcollectionperiod,miceweresuppliedwithriod,sildena?lorvinpocetinewasinjectedinincreasingdoses,aschawandvehicleorsildena?ldissolvedintapwater(acidi?-arterialpressureandrenalblood?owwerepH3).monitoredcontinuouslyusingthePowerLabdataacquisitionsys-Quanti?cationofsodiumbalanceandurinarynitrate/nitritetemandLabChartsoftware(ADInstruments,ColoradoSprings,).(Diet-Gel;Sniff,Soest,Germany)determinedbeforetheapplicationofthe?(%NaCl).?amephotometry(Beckmanofeithersildena?l(800?g/kgBW)orvinpocetine(800?g/kgbodyCoulter),)inacuteexperiments,oraftertheterminationofchronicexperi-balancewasdeterminedbysubtractingsodiumintakefromurinarysodiumexcretion,,renalcortexwascutintosmallpiecesandsnap-,%ice-coldethanolwasmeasuredbyusingacolorimetricassaykit(780001;Caymanusingaglass/glasshomogenizerand?nallycentrifuged(14,000g,15Chemical)andnormalizedtourinarycreatinine(50070;Caymanmin,4°C)anddriedat95°C,andthecGMPcontentwasmeasuredinChemical).-%?SE(n?,andproteincontentwasdeterminedusingthebicinchoninicacidofanimals).Student’st-paremeansoftwogroupsmethod(Uptima).--wayortwo-wayANOVAforrepeatedmea-ofAngII-dependenthypertension,thekidneysofeachgroupweresurements,followedbyBonferroni’parisonposthocisolatedandperfusedwithKrebs-Henseleitbuffer,asdescribedpre-(36).Kidneyswereisolatedfrommiceanesthetizedintraperi-distributionwasnot(orcouldnotbeassumedtobe)normalwereteoneally(ip)withketamine(100mg/kgandxylazine(5mg/kg\).analyzedbytheMann-WhitneyU-??ectedchangesinvascularresistancewereconsideredstatisticallysigni?,abolusof60mMKClwasinjectedtotesttheviabilityofthepreparation,,kidneyswerepreconstrictedwithnorepinephrine(1?M;Sigma-Aldrich).Phosphodiesterase5regulatesbloodpressureandrenalConcentration-responsecurvesinducedbycarbachol(Sigma-Aldrich)blood?-andS-nitrosgluthathione(GSNO;Alexis)wererecordedinthepres-ingrenalblood?ow(RBF)invivo,wetestedtheacuteeffectsenceofeitherdiclofenac(3?M)orL-NAME(300?M;Sigma-Aldrich)anddiclofenac(3?M),?uenceofofsildena?l,aselectivePDE5inhibitor,andvinpocetine,avinpocetine(Calbiochem)andsildena?l(agenerousgiftfromP?zer)selectivePDE1inhibitoronRBFandbloodpressure,underonNO-mediatedvasodilation,weusedasildena?l(300nM)andbaselineconditionsorduringacuteAngIIinfusion(200vinpocetine(10?M)concentrationknowntoreducevasocontractionng·kg?1·min?1).AsshowninFig,1,AandB,sildena?lby?20-30%-?l(80?g/kgrelaxationisexpressedasapercentagepressorresponseofthebodywt:54?9mmHg;800?g/kgbodywt:56?8mmHg),preconstrictedkidney,whichwassetas100%.renalblood?owwasdecreasedinthetwohighestsildena?-infusedWTmice(500ng·kg?1·min?1for14days)weremountedinawireconcentrations(,A–C).DuringAngIIinfusion,PDE5myography(MultiMyographModel610M;DanishMyoTechnol-inhibitionreducedbloodpressureevenmorepotentlyt