- Email: [email protected]
B9 Fibroblast growth factor gene transfer in hypertensive brain vessels
162A
A]H-APRIL 1997-VOL, 10, NO. 4, PART 2
ASH XII ABSTRACTS
B9
B1O
FIBROBLASTGROWTHFACTORGENE TRANSFERIN HYPERTENSIVEBRAINVESSELS, P Cuevas,D Reimers, F Carceller,Q V Barrios::E Asin-Cardiel,G GimenezGaIlego*.HospitalRamony Cajrd,*CSIC,Madrid, Spain Systemic hypertensionis a major risk factor in the dmelopmentof cerebrovascularstrokes,myocardialinfarction and suddendeath.Experimentaldata support a clinicalinterest for fibroblastgrowth factors (FGFs) in the control of cortical cerebral blood flow, brain rm@ogenesisend preservationof endothelialtimction. Furthermore,a progressivedecrease of endothelird FGF accompanies progression of arterial hypertension. A therapeutic approach for comecting brain endothelialFGFs deficiencyis to directlytrrmsfectendothelial cells of the brain vessels with genes encoding FGFs. This strategy would generatean endogenoussource of FGFs from cerebralerrdothelialcells,To teatthe etlieiencyof this approach, humaneeidicFGF (aFGF)recombinantgenewas insertedin the pMAMeFGF expressionplasmid and packaged into catiorric Iiposomes end systemicallyadministeredin 18 month old spontaneouslyhypertensiverats (SHR) (n=6). Control SHR received plasmid pMAMeo (n=6), One month following Iipofectiorr, bloodpressure(BP)was directlymeasured,animals weresacrificedand endothelirdexpressionof aFGF checkedin brain vessels using anti-aFGF immunohistochemistryOnly transected SHR showed a decrease near 300/0in BP in comparisonwith pretreatmentvalues simultaneousto a high expression of aFGF protein in brain vaaculature.This study demonstrateathatdecreasedexpressionofsFGF in hypertensive brain endotheliumcan be correctedby systemicIipofeetionof sFGF gene resultingin long term control of blood pressure. Key words: Growth factor. SH rats. Genetransfer.
HYPOTENSIVE EFFECT OF ACIDIC FIBROBLAST GROWTH FACTOR (sFGF) IS PARTIALLYMEDIATED BY KININS Saenzde Tejsda I, Carceller F, FernandezA, Cuevas B, O&e ~ Gimenez-GallegoG, Cuevss P Dpt. Investigaci6n.Hospital Ramony Cajal, Madrid. Spsin sFGF causes an acute and transient endothelium-dependent decressein bloodpressure in experimentalanimals, However, this response ia not found or is very small in vitro. One possible explanation for this discrepancy is that plaama mediators, perhaps kirrins, are necessary for FGF-induced vasodilation, We studiedthe hyptensive responseto lyg iv aFGF in rats: Group I: control Wistar (n=12); Group 11: trested with a bradykinin receptor mrtsgmriat(Na-adammrtanescetyl-D-Arg[Hyp3, Thi5,s, D-Phe7]-bradykinin) at dosis of 58 pg(n=6) or 420 pg(n=6);Group 111:depleted of kininogen afier repeated injections of cephslin-ellagic acid (4 to 6, 400 @ each, 6-8 min apart; n=5); Group IV: kirrinogen-deficientBNK (n=18),
MeanBP was measureddirectlythrough a femoralcatheter. As comparedwith group I rats the hypotenaiveresponse to aFGF was significantly less pronounced in the reat of the groups, The hypotensiveeffect in group H was reduced in comparisonwith controls by 38% with 58 kg and by 60% with the 420pg dose (9*1 vs 22*3 mmHg;p
Bll
B12
wTOCOPHSROL INCRSASED NITRSC OX3DE tSTNT31AStf ACTIVSTY IN BLOOD VSBSfCI# OF SPONTANEOUSLY NYPERTS!NSIVSRATS(SHR) (x3,hluabm N, GqwrA NawalNNA,MrdatdA Rohaizar Ds@mant of Ricchamis@.~VSdi Keb ~tiem= J* R@ Or Mu& 50300Ku& Lumrrur. hfalmra
ET-1, Cytokines and Norepinephrine Changes After Hsndgrip Test in EaaentialHypertensivea
-
An60xidmrtpmtacden prddad by dmranr doa8s of a-romphamlW’a comrmwtby ddmmink * amthara(NOS) aiivitvin bbwd ems &rR)&tad A a&Ophaml. v~Of&Yh+ SRa Wm dk’idedintofour groups namatyhprtamivs Consml(c), &t tl@mantti 17rsia R-t’3c0pharol,/ks &t (al). 34rrrsCt-t4C0phd& aAm@ramL@diet (@). W&mKyotn(WY) a wem (aZ) md 170nr8 prsmrrs Wam ramrded Svrrr rha Id by usedM llM’MSICOllh_Ol (N). Mood Of ths dudyp8ri0dof ph-h @-ZY ff~ OISdOrrad nmnrha. At ths d of Orstrial, mirnsrs WereSminratmrdNOS C#iewavsalsay acdvity in bloodvaaaehm ~ w ml+ White nitritaconcmtraion ~-~ ~ * ~ 554nmuairrg OrE&sMagarrt. Analysis of &a m ti mrdanss’ t-tadfor dlmance. ThsCmnputar pqnmrms STATIST3CAm uad for au maryai,.Raanlts of our msdydurwadmmfor au ths thraa& (p3J3ee) ~1 WrrPs. ~ ~ arrdmaximum MiucdOnof I-adMad Cmnpmdto thahypltcm“W COMd btoodfmuurs WM ahmm by OIS34rnsaaqrhaml RS diat (G209.5d+S.47rnrn~ @12R8?ck17,13nmr 3fa), Aim , NOSwthdtyin towar OranWKY m Mood Vc6a3sof SNR Wm S@dam@ (WKY=l.54M26pnr0unrgpmtaa aHR=e.e7+33mprmvmg pnxehu p=e.eq. Anar h—wmmt. duroughWoqhamr dosraof a.z md U3 ~ ChaNOS ill btOOdVm .X@ ti Of ti anR=e.e7Mu3 mgmnace (oz=l.1e2L0.zapmnI& m@rlcmuy Pnmrtnra fnutain; P=m4). Prunrra rdldss~ ruduradin SNR mnqmed to WY ti (WKY=54,6M2,%jnneMr& s4m=26.24M.14prnolm p=e.ee)d ScWnS@y arttfnmfmsps Showed inoleirmpsdrverdhita tamr(al: p=e.eees,.32: p=e.tweaLY3:pe.eez).Irlcmcbwionitvr6f0rrrrdr3aammpa’ad to WKY ma , SHRha towerNOSactiv#yinbIced whichqxm tra#rnW ths NOS artM5y md ~~brr-t with Wkxidmt ~~ psurrre.TldabrytieJthd SW Imtimra &ggg& ~ ~ ~ of~ Key words:
GTec+ravL Mtric mida aynthm, Nikite, Prae m3isslhmedPraaura.
S.Cottone A.Vadal& MC Vella, G.MuI&,R.Riccobene, —! andG.Cerasola*; (lair oflntemal Medicineand HypertensionCenter;Universityof Palermo,Italy The interactionof several vaaoactivesubstances such as catecholamines,cytokines,and endothelinsplays an important modulatingrole in hypertensive disease.Nevertheless, many aspects of this interactionare not clearly understood. In order to analyze the behaviour of serum Errdothelin-l(~l),Fibroblast- (FGFI and Platelet Derived Growth Factors (PDGF) , Tumor Necrosis Factor (TNF) , and of plasma Norepinephrine (NE) both in basal condition and after handgrip test (performedfor3 rein) in essential hypertension (EH), we studied 15 mild EH with mean age 43t2 yrs and 8 age-matched normotensive controls (C). In basal condition, EH showed higher values of FGF (14.2~0.5 vs 9.~0.4 pg/ml, pcO.001) and of NE(462~ 8.4 vs 30G~ll pghnl) fhrrrrC , whereas circrdatirrgEl-l, TNF and PDGF were quite similar. 1nEH after handgrip test there was a signiticantincreaae in~-1 (KO.002), FGF (~ 0.006), TNF (pcO.01) and NE (pcCr.0005) as well as in systolic ([email protected]) and diaatolic(pcO.lXG5) blood pressures. In C the increases in ET-1 and cytokines after handgrip test did not reach the statisticalsignificance. In EH serrrmFGF both in basal condition and after handgrip
test significantlycorrelatedwith ET-1 (r 0.67 and 0.50, pc 0.01 and 0.05; respectively),and FGF mm-elatedwith SBP (r 0.48,w0.05) after handgriptest only. No correlationsof ET-l withNE nor with bled pressureawerefound. Theseresrdtaaee.mto indicatethe existenceof an activation of cytokinesisrEH. Furthermore,cytokinesseemto be closelyrelatedwithvascularstress and with ET-1.
Key Words:Ertdothelin,Cytokines
A]H-APRIL 1997-VOL, 10, NO. 4, PART 2
ASH XII ABSTRACTS
B9
B1O
FIBROBLASTGROWTHFACTORGENE TRANSFERIN HYPERTENSIVEBRAINVESSELS, P Cuevas,D Reimers, F Carceller,Q V Barrios::E Asin-Cardiel,G GimenezGaIlego*.HospitalRamony Cajrd,*CSIC,Madrid, Spain Systemic hypertensionis a major risk factor in the dmelopmentof cerebrovascularstrokes,myocardialinfarction and suddendeath.Experimentaldata support a clinicalinterest for fibroblastgrowth factors (FGFs) in the control of cortical cerebral blood flow, brain rm@ogenesisend preservationof endothelialtimction. Furthermore,a progressivedecrease of endothelird FGF accompanies progression of arterial hypertension. A therapeutic approach for comecting brain endothelialFGFs deficiencyis to directlytrrmsfectendothelial cells of the brain vessels with genes encoding FGFs. This strategy would generatean endogenoussource of FGFs from cerebralerrdothelialcells,To teatthe etlieiencyof this approach, humaneeidicFGF (aFGF)recombinantgenewas insertedin the pMAMeFGF expressionplasmid and packaged into catiorric Iiposomes end systemicallyadministeredin 18 month old spontaneouslyhypertensiverats (SHR) (n=6). Control SHR received plasmid pMAMeo (n=6), One month following Iipofectiorr, bloodpressure(BP)was directlymeasured,animals weresacrificedand endothelirdexpressionof aFGF checkedin brain vessels using anti-aFGF immunohistochemistryOnly transected SHR showed a decrease near 300/0in BP in comparisonwith pretreatmentvalues simultaneousto a high expression of aFGF protein in brain vaaculature.This study demonstrateathatdecreasedexpressionofsFGF in hypertensive brain endotheliumcan be correctedby systemicIipofeetionof sFGF gene resultingin long term control of blood pressure. Key words: Growth factor. SH rats. Genetransfer.
HYPOTENSIVE EFFECT OF ACIDIC FIBROBLAST GROWTH FACTOR (sFGF) IS PARTIALLYMEDIATED BY KININS Saenzde Tejsda I, Carceller F, FernandezA, Cuevas B, O&e ~ Gimenez-GallegoG, Cuevss P Dpt. Investigaci6n.Hospital Ramony Cajal, Madrid. Spsin sFGF causes an acute and transient endothelium-dependent decressein bloodpressure in experimentalanimals, However, this response ia not found or is very small in vitro. One possible explanation for this discrepancy is that plaama mediators, perhaps kirrins, are necessary for FGF-induced vasodilation, We studiedthe hyptensive responseto lyg iv aFGF in rats: Group I: control Wistar (n=12); Group 11: trested with a bradykinin receptor mrtsgmriat(Na-adammrtanescetyl-D-Arg[Hyp3, Thi5,s, D-Phe7]-bradykinin) at dosis of 58 pg(n=6) or 420 pg(n=6);Group 111:depleted of kininogen afier repeated injections of cephslin-ellagic acid (4 to 6, 400 @ each, 6-8 min apart; n=5); Group IV: kirrinogen-deficientBNK (n=18),
MeanBP was measureddirectlythrough a femoralcatheter. As comparedwith group I rats the hypotenaiveresponse to aFGF was significantly less pronounced in the reat of the groups, The hypotensiveeffect in group H was reduced in comparisonwith controls by 38% with 58 kg and by 60% with the 420pg dose (9*1 vs 22*3 mmHg;p
Bll
B12
wTOCOPHSROL INCRSASED NITRSC OX3DE tSTNT31AStf ACTIVSTY IN BLOOD VSBSfCI# OF SPONTANEOUSLY NYPERTS!NSIVSRATS(SHR) (x3,hluabm N, GqwrA NawalNNA,MrdatdA Rohaizar Ds@mant of Ricchamis@.~VSdi Keb ~tiem= J* R@ Or Mu& 50300Ku& Lumrrur. hfalmra
ET-1, Cytokines and Norepinephrine Changes After Hsndgrip Test in EaaentialHypertensivea
-
An60xidmrtpmtacden prddad by dmranr doa8s of a-romphamlW’a comrmwtby ddmmink * amthara(NOS) aiivitvin bbwd ems &rR)&tad A a&Ophaml. v~Of&Yh+ SRa Wm dk’idedintofour groups namatyhprtamivs Consml(c), &t tl@mantti 17rsia R-t’3c0pharol,/ks &t (al). 34rrrsCt-t4C0phd& aAm@ramL@diet (@). W&mKyotn(WY) a wem (aZ) md 170nr8 prsmrrs Wam ramrded Svrrr rha Id by usedM llM’MSICOllh_Ol (N). Mood Of ths dudyp8ri0dof ph-h @-ZY ff~ OISdOrrad nmnrha. At ths d of Orstrial, mirnsrs WereSminratmrdNOS C#iewavsalsay acdvity in bloodvaaaehm ~ w ml+ White nitritaconcmtraion ~-~ ~ * ~ 554nmuairrg OrE&sMagarrt. Analysis of &a m ti mrdanss’ t-tadfor dlmance. ThsCmnputar pqnmrms STATIST3CAm uad for au maryai,.Raanlts of our msdydurwadmmfor au ths thraa& (p3J3ee) ~1 WrrPs. ~ ~ arrdmaximum MiucdOnof I-adMad Cmnpmdto thahypltcm“W COMd btoodfmuurs WM ahmm by OIS34rnsaaqrhaml RS diat (G209.5d+S.47rnrn~ @12R8?ck17,13nmr 3fa), Aim , NOSwthdtyin towar OranWKY m Mood Vc6a3sof SNR Wm S@dam@ (WKY=l.54M26pnr0unrgpmtaa aHR=e.e7+33mprmvmg pnxehu p=e.eq. Anar h—wmmt. duroughWoqhamr dosraof a.z md U3 ~ ChaNOS ill btOOdVm .X@ ti Of ti anR=e.e7Mu3 mgmnace (oz=l.1e2L0.zapmnI& m@rlcmuy Pnmrtnra fnutain; P=m4). Prunrra rdldss~ ruduradin SNR mnqmed to WY ti (WKY=54,6M2,%jnneMr& s4m=26.24M.14prnolm p=e.ee)d ScWnS@y arttfnmfmsps Showed inoleirmpsdrverdhita tamr(al: p=e.eees,.32: p=e.tweaLY3:pe.eez).Irlcmcbwionitvr6f0rrrrdr3aammpa’ad to WKY ma , SHRha towerNOSactiv#yinbIced whichqxm tra#rnW ths NOS artM5y md ~~brr-t with Wkxidmt ~~ psurrre.TldabrytieJthd SW Imtimra &ggg& ~ ~ ~ of~ Key words:
GTec+ravL Mtric mida aynthm, Nikite, Prae m3isslhmedPraaura.
S.Cottone A.Vadal& MC Vella, G.MuI&,R.Riccobene, —! andG.Cerasola*; (lair oflntemal Medicineand HypertensionCenter;Universityof Palermo,Italy The interactionof several vaaoactivesubstances such as catecholamines,cytokines,and endothelinsplays an important modulatingrole in hypertensive disease.Nevertheless, many aspects of this interactionare not clearly understood. In order to analyze the behaviour of serum Errdothelin-l(~l),Fibroblast- (FGFI and Platelet Derived Growth Factors (PDGF) , Tumor Necrosis Factor (TNF) , and of plasma Norepinephrine (NE) both in basal condition and after handgrip test (performedfor3 rein) in essential hypertension (EH), we studied 15 mild EH with mean age 43t2 yrs and 8 age-matched normotensive controls (C). In basal condition, EH showed higher values of FGF (14.2~0.5 vs 9.~0.4 pg/ml, pcO.001) and of NE(462~ 8.4 vs 30G~ll pghnl) fhrrrrC , whereas circrdatirrgEl-l, TNF and PDGF were quite similar. 1nEH after handgrip test there was a signiticantincreaae in~-1 (KO.002), FGF (~ 0.006), TNF (pcO.01) and NE (pcCr.0005) as well as in systolic ([email protected]) and diaatolic(pcO.lXG5) blood pressures. In C the increases in ET-1 and cytokines after handgrip test did not reach the statisticalsignificance. In EH serrrmFGF both in basal condition and after handgrip
test significantlycorrelatedwith ET-1 (r 0.67 and 0.50, pc 0.01 and 0.05; respectively),and FGF mm-elatedwith SBP (r 0.48,w0.05) after handgriptest only. No correlationsof ET-l withNE nor with bled pressureawerefound. Theseresrdtaaee.mto indicatethe existenceof an activation of cytokinesisrEH. Furthermore,cytokinesseemto be closelyrelatedwithvascularstress and with ET-1.
Key Words:Ertdothelin,Cytokines