La lecture à portée de main
Découvre YouScribe en t'inscrivant gratuitement
Je m'inscrisDécouvre YouScribe en t'inscrivant gratuitement
Je m'inscrisDescription
Sujets
Informations
Publié par | biomed |
Publié le | 01 janvier 2009 |
Nombre de lectures | 8 |
Langue | English |
Poids de l'ouvrage | 1 Mo |
Extrait
.3eRniohdlU_bmruchvorlage28.11.0910:03Seite520520
EuRoPEanJouRnalofMEdIcalRESEaRcH
december14,2009
EurJMedRes(2009)14:520-525©I.HolzapfelPublishers2009
R
EnIn
,E
ndotHElIal
noS
yntHaSEand
E
ndotHElIn
G
EnE
E
xPRESSIonIntHE
2K
IdnEy
-1
clIP
G
oldBlatt
M
odElof
l
onG
-
tERM
R
EnovaSculaR
H
yPERtEnSIon
S.W.Reinhold
1
*,d.c.uihlein
1
*,c.a.Böger
1
,S.Kloiber
1
,K.frölich
1
,t.Bergler
1
,B.Banas
1
,
f.Schweda
2
,B.K.Krämer
3
1
Klinikun
2
dInPstoiltiuktlifnüikrfPühryIsinonleorgeie,MeudniizvienrsIiIt,yuonfivReergseitnysbofurRg,egReengsebnusrbgu,rRg,egGeenrsmbuarngy,,Germany,
3
MarienhospitalHerne,Ruhr-universityBochum,Germany
bstract
Objective:
numerousreportshaveshowntheinfluence
ofrenin,nitricoxide(no)andtheendothelin(Et)
systemsforregulationofbloodpressureandrenal
function.furthermore,interactionsbetweenthese
peptideshavebeenreported.aimofourstudywasto
investigatetherelativecontributionofthesecom-
poundsinlong-termrenovascularhypertension/re-
nalischemia.
Methods:
Hypertension/left-sidedrenalischemiawas
inducedusingthe2K1c-Goldblattratmodel.Renal
renin,Et-1,Et-3andendothelialnosynthase
(enoS)geneexpressionwasmeasuredbymeansof
Rnaseprotectionassayatdifferenttimepointsupto
10weeksafterinductionofrenalarterystenosis.
Results:
Plasmareninactivityandrenalreningeneex-
pressionintheleftkidneywereincreasedinthe
clippedanimalswhileenoSexpressionwasun-
changed.furthermore,anincreaseinEt-1expression
andadecreaseofEt-3expressionwasdetectedin
earlystenosis.
Conclusions:
Whilereninisobviouslyinvolvedinregu-
lationofbloodpressureandrenalfunctioninunilater-
alrenalarterystenosis,Et-1,Et-3andendothelium
derivednodonotappeartoplayanimportantrolein
renaladaptationprocessesinlong-termrenalartery
stenosis,althoughEt-1andEt-3mightbeinvolved
inshort-termadaptationprocesses.
Keywords:
bloodpressure,reninangiotensinsystem,
nitricoxide,endothelins,long-termrenalarterysteno-
sis,Rnaseprotectionassay
I
ntRoductIon
therenin-angiotensinsystem(RaS)hasgenerally
beenacceptedtoplayanimportantroleinbloodpres-
surehomeostasis.ReninformsangiotensinIfroman-
giotensinogenandangiotensinIisthereaftertrans-
*Bothauthorscontributedequallytothispaper.
formedbytheangiotensinconvertingenzymeinthe
biologicallyactiveangiotensinII(at-II).at-IIis
knowntobevasoconstrictiveandstimulatesthere-
leaseofaldosterone,therebycausingsodiumreten-
tion.theupregulationofrenininrenalarterystenosis
duetotheactivationoftherenalbaroreceptoris
thoughttocauserenalhypertensionbyactivatingthe
renin-angiotensinsystem.
noisknownasvasorelaxingmolecule.Inthemod-
elof2K1crenovascularhypertensionnohasbeen
proventobeinvolvedinmaintainingbloodflow.no
isthoughttoactasavasodilatatorandtherebytoan-
tagonisethehighlevelsofat-IIinbothkidneys[1].
nomeasurementshaveshownthatendogenousno
productionisincreasedinthecontralateralkidneyof
2K1cratsthreeweeksafterclipping[2].
theEtscompriseagroupofthree21-aminoacid
peptides,havingvasoactive,inotropicandmitogenic
properties[3].Et-1actsonvascularmusclecellscaus-
ingalong-lastingvasoconstriction[4].Short-termrenal
arterystenosisinthe2K1cmodelofrenovascularhy-
pertensionhasresultedinanupregulationofrenalEt-
1intheclippedkidney.Et-2wasnotfoundinratkid-
neys[5].furthermoreEt-1andEt-3seemtoberegu-
latedinoppositewaysasshowninaratrenalischemia
reperfusionmodel6hoursafterreperfusion[5].
Inaddition,manyinteractionsbetweentheseabove
compoundshavebeenfound:noforinstancedown-
regulatestheEt-1expressionandat-IIisaknown
stimulatorofEt-1expression[6,7].
Sincethecontributionofthesedifferentmediators
inrenovascularhypertensionespeciallywithregardto
long-termregulationisunclear,theaimofourstudy
wastoinvestigatetheeffectsofunilateralrenalartery
clipping,usingthe2K1c-model,onrenalrenin,
enoS,Et-1andEt-3geneexpression.
M
EtHodS
allanimalexperimentswereconductedaccordingto
thenationalInstitutesofHealthguidelinesforthe
careanduseofanimalsinresearch.
3.Reinhold_Umbruchvorlage28.11.0910:03Seite521december14,2009EuRoPEanJouRnalofMEdIcalRESEaRcH521
80maleWistarrats,weighing150g,havingfreeac-B
lood
P
RESSuRE
cesstotapwaterandstandardcommercialpelletchow
(altrominc1000,lage,Germany)wereused.unilat-Systolicbloodpressuresignificantlyincreasedinthe
eralrenovascularclippingwasinducedinatotalof402K1c-groupto162.5±8mmHgonday1,179±11
ratsandashamoperationin40controlrats.asteno-mmHgonday2and189±7mmHgonday20after
sisoftheleftrenalarterywasinducedbya0.2mmsil-clipping,thereafterlevellingoffatslightlylowerlevels
verclip.anaesthesiawasdoneusingmethohexital(75(155±7mmHgatday40and149±3mmHgatday
mg/kgIP).8animalsofeachgroupweresacrificed3,63).thecontrolgrouppresentedsignificantlylower
7,12,28and70daysaftersurgery.systolicbloodpressurevaluesateverypostoperative
Systolicbloodpressurewasmeasuredusingthetailtimepoint(fig.2).
cuffmethod(processcontrolbloodpressure209000;
tSE,Homburg/Saar,Germany)[8].Bloodwascol-
lectedformeasurementofplasmareninactivityafter
decapitation.thekidneyswereimmediatelyremoved,
weighedandfrozenusingliquidn
2
andstoredat
-80°c.
Renalrenin,enoS,Et-1and-3and-actingene
expressionfromwholekidneywasanalysedusing
Rnase-protectionassaysaspreviouslyreported[9].
-actinwasusedashousekeepinggene.Plasmarenin
activitywasmeasuredbyacommerciallyavailable
anG-I-radioimmunoassay(diasorin,Saluggia,Italy).
Resultsareexpressedasmean±SEM.theStu-
dent´st-testwasusedforstatisticalanalysisaftertest-
ingfornormaldistribution.Pvalues<0.05werecon-
sideredstatisticallysignificant.
R
ESultS
K
IdnEy
W
EIGHt
Kidneyweightoftheunclippedkidneysincreased
Fig.2.
Systolicbloodpressureafterclippingandsham-opera-
bytime.Itwassignificantlyhigherthantheclipped
tion.Systolicbloodpressuresignificantlyincreasedinthe
2K1c-groupto162.5±8mmHgonday1,179±11mmHg
kidneysstartingatday7.thecontralateral(unclipped)
onday2and189±7mmHgonday20afterclipping,there-
kidneyhadahigherweightcomparedtothecontrol
afterlevellingoffatslightlylowerlevels(155±7mmHgat
groupstartingatday28.theweightoftheclipped
day40and149±3mmHgatday63).thecontrolgroup
kidneyswassignificantlylowerthancontrolstarting
presentedsignificantlylowersystolicbloodpressurevalues
atday3anddidnotchangesignificantlyovertime
ateverypostoperativetimepoint(significantvs.control*,
(fig.1).
p<0.05).
Fig.1.
Kidneyweightinthe2K1cgroupandcontrolgroupatdifferenttimepointsaftersurgery.Kidneyweightoftheun-
clippedkidneysincreasedbytime.Itwassignificantlyhighercomparedtotheclippedkidneysstartingatday7.thecontralateral
(unclipped)kidneyhadahigherweightcomparedtothecontrolgroupstartingatday28.theweightoftheclippedkidneyswas
significantlylowerthancontrolstartingatday3anddidnotchangesignificantlyovertime(significantvs.control*;significant
vs.contralateralside#,p<0.05).
3.Reinhold_Umbruc225
vhorlage28.11.0910:03Seite522EuRoPEanJouRnalofMEdIcalRESEaRcHdecember14,2009
Fig.3.
Plasmareninactivityintheclippedandunclipped
group.Plasmareninactivityinthe2K1c-groupwassix
timeshigherthaninthecontrolgroupatday3.Plasma
reninactivitythendecreasedtoabout3.5foldondays7,12
and28andtoabout2foldofcontrolgrouponday70(sig-
nificantvs.control*,p<0.05).
Fig.4.
Geneexpressionofreninintheclipped
andunclippedkidneys(K)atdifferenttime-
pointsaftersurgery.Renalreningeneexpression
washigheratalltimepointsintheclippedkidney
comparedwiththeunclippedkidneyandatday
3to28comparedtocontrol,whereasthecon-
tralateralkidneydisplayedreningenesuppres-
sionto~50%ofcontrolatday3to12(signifi-
cantvs.control*;significantvs.contralateral
side#,p<0.05).
Fig.5.
GeneexpressionofEt-1intheclipped
andunclippedkidneys(K)atdifferenttime-
pointsaftersurgery.Et-1geneexpressiontend-
edtoincreasewithtimeinallgroupsandwas
significantlyhigherintheclippedkidneyonday
7comparedwiththecontralateralkidney(signifi-
cantvs.contralateralside*,90.4±6.7vs.71.2±
5.5,p<0.05).
P
laSMa
R
EnIn
a
ctIvIty
clippedkidneyandatday3to28comparedtocontrol,
whereastherightcontralateralkidneydisplayedrenin
Plasmareninactivityinthe2K1c-groupwassixtimesgenesuppressionto~50%ofcontrolatday3to12
higherthaninthecontrolgroupatday3.Plasmarenin(fig.4).
activitythendecreasedto~3.5foldondays7,12and
28andto~2foldofcontrolgrouponday70(fig.3).R
Enal
Et-1G
EnE
E
xPRESSIon
R
EnIn
G
EnE
E
xPRESSIon
Et-1tendedtoincreasewithtimeinallgroups.fur-
thermoreEt-1geneexpressionintheclippedkidney
Renalreningeneexpressionwashigheratalltime-washigheronday7comparedwiththecontralateral
pointsintheclippedkidneycomparedwiththeun-kidney(90.4±6.7vs.71.2±5.5;p=0.046)(fig.5).
.3eRniohdl_Umbruchvorlage28.11.0december14,2009
19:030eStie523EuRoPEanJouRnalofMEdIcalRESEaRcH
R
Enal
Et-3G
EnE
E
xPRESSIon
Et-3tendedtoincreasewithtimeinallgroups.Et-3
geneexpres