Rostamzadeh, Farzaneh

Simorgh Research Repository

-.4-{ryltri4't"UL.l,r{ ,,J,iit ,*S,:i olS,i,it.rJcsiJJJlp gti.3-:5-xcEjols:;+ (phD) .,.-,-sl <sl;X, &r- -U ,L!:;l.rcol;l ;.r,*l 6La o.r.=,K tr tr;T &t :, !K,s+$.x*l 6La oa.,K oii: .,-__-.13,-: 63Liii + q lt *- ; 6rlJ--- ,-i_l- :*S t.l agl*l ,ji.o _, .-, jl ^ ,S#3; :-dr--r. t5g$ort3 f-, +.l-i-;s , L....-F-l,l* ,r+i 4-'"- ;iS.l :L"^l., ll':"'tdL^L +r*., _!S, -;tS.n ,,i_l!,.., tiS.l :r3l .L o .ll'1. ..'l\Y11:.Jf,--: Jl,-,I\lUa,{rf'' -il"tugKerman University of Medical SciencesFaculty of Medicine and Physiology Research CenterIn Partial Fulfillment of the Requirements for the Degree of Ph.D in MedicalPhysiologyTitle:Role of kapa opioid receptors and their interaction with apelinreceptors in positive and negative inotropic effects of apelin in theheart of rats with coronic renovascular hypertensionByFarzaneh RostamzadehSupervisorDr. Hamid NajafipourAdvisorsDr. Syavash Joukar - Dr. Saeed Esmaili MahaniYear: 2017o+5?.'.rj!.r Gra 9,fi.St^ 4:5-,.^i! af,,l, &!F9 i:l29.l ,rLl p.> ,b1* ;\,ts rf cS,r[$,r, :a"rl.51-o91lr .t1.t i tu:6 r.l*,l1L*-, ;.l .,r*i! u" *.yV c,f 6s$x 3-L; -|,_Y: jl ,F_ sy$ 4f 1!;',-...1Jl .-ro:.f ,&5 S# of 6sL$y, "L;;l c-t1o ,js6X,,, 1,, .tL-,,- tm.t*lr.l ;9 7L;-i o-*-(APJ) JJI .gto o$;5.s).r",rr-lil l) -J-i s,-lr5 A:f Sl; ro.J:*. * i\ l;)i e :t e.b* yl U;g*Jtel rlti ,: (KORs) J+ie.l !.1J gto "sr5 aL-' ,l (GPCRs) G .r."'ix L o,u c-o> .51-o orj75,'-- -ERK1/2 .r9*.)4,-rr,*s 9 C jqt ,-yqsx alo rl ,rr!>.- gb,"* jl J+,1 .s;1.: a,!2t 9: r599;i,5 6)yr.*,j9.-\/o.,,s 9 PKC .lr; Jti ;lr*" KORs 9 APJ rrr.-"li)--)r,p .ra) .f .;-1lpl 1; eG .5;4 .,-*.c-l a;L;L.rJ,,l .p:f 9.r*ti o1l1 ,,r,-,*!,1 g s:ix|.5b o$;,5 J"tJ ,J;j.s) cr, "i,.l;l 1) ERK, -es-- s of .s,,Hr ;l J-r. y_l ,,, .-,*L-1 ,rJ!r olll ,.: ERK1/2 rr9.-\)ar I C ;t5 ,-ytrsy, "*jij ;l ,',J/- $1, .St",5 *7,L ,p-l ,_& r-l- allt-.s-r.o;.1r,[ ,colo-u] d:;l-i-i d+"j ar-lt:.S$Xlcilo .- -'0s."5 .-,* -l.U I J9,.*11_)--)rr* .,lS ,r .xtl ,1U ,KOR , APJ Jr*"1r.)^,_)9;:o ,KORs .51-o ol,;i _ -.rysy[ c1"f e-rp.s,,S,r.J-,,r._,*l ;irr*-$oi;l syAloiF sc5S; -,-J* .rf cs,,S.r .s rh;l i,5 YY. b \4. ds\ ;\4.g:1; ; u,lr>-- 09" \2^ 69) .t AJtt, r,l :J;.u'ly-.yf :.t d.r!.f cSSt p *l* ./Y ,.I"s !.,^,W *-"(J," ""1t.t" drul*t \ Sl "F..(,-,_5'.- *.,-lad;1.r". ( O^f Ls)l";i)L:l-a,l) -^J5 d;t lrtrl ,*" a:.o,o \,.r,r; p*-d',rrlr A og.F Y\ a, ot,l9;.-;*. + t+7, +;V9(l.j !_ri r:"G !+.Jh,Jl,,rr r!.,"".,, e-,,b,rt" t? os.ts ,-tl - p.r.rr _-t *+ 9 G\./..it JyK .r; a;:;[f :), e;y: .*i6 .s;1,- .r-!iil .glo ,aL" ,.S-Li.:ra .Slt,,i5G c+if il a;t,-y..b ;l?. :f .pg/kg dbjr) li.r*1r.1 .$r; J.-i" aJUJA r-irt!. *r-..- + L-'r., t rl* ,uy.t;t';r-Iot-r(.r*Jxl .slooJ;,5.ff cr.rl5)l4d) JLJrL,(oJ.l ;6 osf S.r5,,kr) F13A.q)f ,-y.;.-o:lo;l e,':U ,-y.ti )l J., or*, .: (PKC ".5 r\r)Chelerl'thrine L- 9 ( !.15 .rl^ oJ;,5 cs,li:";l .ut)l{d)B\-.s!.K .s ol..f ,-y::sl- 9 mRNA oV_ SF orlsl .:-; ,-t-,,i ..,J.1 ,l -f,s ;.cL fA (Gi .#,,\r)PTX .;-,y:;\ LK "r;*< !:*,,1 ,*,f ;y-"i,*-J.! rb-.. -){-' ,:l-l: Real Time PCR t..+'i { G4-.:,,:rs u;\.);l l;Yl jel ! ;i ,r-\.N s ERK 1/2 )l-r.,L .r; cs.;N eyry;-"e-'p-J- \V\/\Yf e \ \' lY\ qi q 2K1C e p; ,ef ,: !pt-:/tf*- ;t-)- ,L;-i ;JiL :61tr;*!l olr,lnor-BNl .sr.5,,hr Y. s2. pg/kg jtr l):b rr l, ;^Ll ,L;-i..rii"-ol5 olll ;tJlU 9 F13.\t. J*1,,.1il9 PTXt-y;t-:rrG,rf. J"lr,lr,l .rlr";otl 2. ;f,,):,,\,.y.ttf, l9rrr1reL_r,--..:1r,;-lrsl lr:-M;1,-, eVlY, l7'_u)l),,".,).-,*!.1 .!-),5.,l4,"J.r-,J-9PTX L,y G\.r,--nor- .$ )t{" PTX ! : ;!!. .i*tt *.x/- 9 nor-BNI ,,jtrrL ,F13A ! ."1, .s1,, jl) t) ra )) e"- -.5Lu osrS ,lt,;*o ,Qr .ss t. J+.1 t-9, -clp/dtnrax , +dp/dtmax c"il ;lr> jl eL uxF :+ &_!. r.: l, -^Jji 6t.t- eVl , r;ol5 1., ;L-l ,b,._'J.l 9 ts:i9e-l Lll .srS Jt,;'l rhr I uJ' ,9 "p.l.pl 1, KORs 9 APJ Jy..-1i.F)9yb g S:,J-g*,I Lt5 .sr5 ,-ytrSy..,L .-,1' g.,S,r, ,.:b ,-, -- 'a,, l, ERK1/2 Jy-"\l P,s s oy!,ly.n.:9;a ,l-r; jl) .l) ra ,,) ,-.,J,1 .:l: ,r;ol5 l) ERKI 12 ul=.>'- "";l-r--,J- .rl: ,;_lrsl l) t.D+l { e! ,: ERK1/2 rr}-\,-r .SIJXI Ltl os;5 r\a .sl:.i .-l)?, ;.!l { e"! )) ERK1 12 Jy->t".P}, PTX 9.,\rl, o*l jl) r) p { e-.t?.rr ERK1/2 Jy"\*-:.:h ,:,o15c"'L APJ I cs{xl .56 o$;5,t-u; .r5 e 6.F.,2.j'y-1, oJ.l 6*;i ,_*,1 s 6*-,:iyl..1olil , .:,: !.tl .rt" os;.f ;L ,";2K1C ;f glbr .)9., .r" -*J,l ;;i ,J*-3isr.l ll l,,S .5*lJ ol;l -r,-:*o1;1 atl; ERK1/2 ,i^5 \i j9:.,1i.)"r.)r;^a .rnl,'-i .s) -l .r--l,pl l, .r,r5 l) .l"l .-,r-lir--r,..*^2) ,e^I! rrsJ^.c Jy-l .Jjly.rp9,!..,),:-l .;S^. ol2*+ 6,1 .ro; e /....-1, .r*l.l J*,ii ,s"l s S*"gy>ERK1/2 r[L rr*a..............,p J+l il .*;t?,f^i:,c-1 ,*,1 rr- &.r^tS !.1,* $ ,r"r" s# ,f Gt\;.-.s,,H,*, )l Ly ,.y.|r:;Lr: i-u.i- ,l:- tlr.-,*t_l s1t 4 KORs 9 APJ Jy--1i)4)9)-i JlF 9 4L-,.-.s eb -:CrUJ *isr,ot-F*,! ERKl/2 ,*.[i .s,p-!. eV',c5$ry.1 llJ "r;r5 ,grl*"1 .glo "-r"75 ,.*lil or;-< ,rSyY c;p .J.,bX ,rf*,1i2^,-ri7,.o :dJ+15 gtor\bstractsIntroduction: Il.vpertension is one o{'the main risk I'actors fbr the development and--,''gression ol.heart lailure. rn1'ocardial inf-arction (MI) and Stroke. Renovascular hypertension: rrre of the major types ol'secondar,v hlrpertension. T'hese patients are prone to resistance to. rod pressure reclucing drugs. Apelin clccreaseci the artcrial pressure in hyperlension including'--:ovascular hypertensior-r. As apelin/APJ svstcm inr,'oil'es in the regulation of blood'r'.'ssure/myocardial contractility. it is not surprising that the apelin/APJ systen-r plays direct and...Jirect roles on the function of the heart. Apelin receptors (APJ) cross-talk w'ith other G-protein-- -Lplecl receptors such as kappa opioid receptors (KORst on ph,vsiologic functior-rs. Apelin has-.,srtrve cardiac inotropic elfects through protein kinase C (PKC) activation and phosphorylation- ERKi/2 and the heterodimerization of APJ and KORs increases PKC activation and.',sphor)'lation of ERKli2 in response to apelin. The role of interaction between opioid and::lin reccptors on cardiovascular ef'fects of apelin uas not knowr. Therefbre. the present study.rccl at evaluating the heterodimerization betu,een KOR ancl APJ and assessing the impact of', ancl high closes o1' apelin on the lel'el of heterodimerization in the myocardium in- -,\i:lsclllar ir1-perter-rsion. In addition. as the role'ol EIIKli2 phosphorylation and PKC in::iatir-rg the ctrrdiac intpacts of apelin in rats rvith renovascular hypertension and the role of..1 receptors (OPRs) in this regard is unclear. the rolc of these intracellular n"rediators. the- . .:ssior-r of KORs. and tl-re role of intcraction betlveen OPRs ancl APJ on the cardiovascular- -:JlS of apelin in a chronic renor,'ascular rnodel of hypertension was also investigated.\lethods: Erperiments nere carried oLlt on 168 Wistar rats u'eighing 170- 200 grams. For-,,..trn of chror-ric 2K1C hy'pertension. after ar"reslhesia rvith ketamine and xylazine a 0.2-ntn.t. , .:,;'r pleriulas clip was placed on the leI1 kidney arter,v. The animals were divided into tu'entl'- -:-,'.rps n'ith 8 rats in each. At the end of week 16. afier anesthesia with sodium thiopental.'-:.r catireters tllled rlith heparin saline r,r,ere placed in right femoral afier)', left r'entricle 1L-\'1., ,r :ire jugular r.ein tbr recording hemodl'namic anci cardiac contractility incidences and iniectionlhe drugs respectivelv. 'l'he artery anc'l vcrrtricular cannulas rvere connected to pressure-'-.rsducers and then to an 8-channel Pou.erlab svstem.. .3A (an APJ antagonist), naloxone (a general OPR inhibitor). nor-BNI (a selective inhibitor ofiOR) and chelervthrine (PKC inhibitor) were given IV l0 rr-rin prior to injectior-rs of apelin atioses of 40 and 60 pLgr'kg. PTX (Goi inhibitor) was in-jected intra-peritoneally' 48 hours prior toihe hemod-vnamic experiments. 'l'he heteroclimerization of KOR and APJ. the expression of KORntRNA and protein in the lefi ventricle of 2KlC rats wcre measurred b.v immunoprecipitation.nestern blot and. real time PC|R respectivell'. 'fhe levels of ERK1/2 phosphorylation n'erer'r'aluated by F.l-l SA.Results: The arterial s1'stolic/diastolic blood pressure in sham and 2K1C rats w'as 110/71 mmHg,.rtd 171/124 nrrnHg. respecti\elr'. The h1'potensive ef-fects ol.apelin at both doses were inhibited:r F134 and naloxone. Nor-BNI completel.v inhibited the ellbcts of apelin 40 on arterialpresslrre..,nd decreasecl the eI-lects at 60 pg/kg. Apelir-i ,10 ar-rd 60 decreased blood presslrre through Gi:athway. Apelin 60 also rcduced blood pressure through PKC pathw'a.v. Apelin in both doses.ncreased cardiac contractility. The cardiaic impacts in both closes w'ere reduced by F'13A.:',rlorone. nor-LINI ancl cl-relerl'trine and blockeci b1, P'fX. KORs inhibition and chelerl,trine also:revented the compensation fbr the decrease in tl.re left ventricie +dp/dt max and -dp/dt max,.tr"tsed b1' apelin 60. 'l'iic sirnultaneous inhibitii>r.i ol'OPR and APJ and KORs ar"rcl APJ reduced-,r'terial pressure anc'l increased cardiac contracttlitf in response to apelin 60. Hl,pertension.:creased the expression of KOI{s and heteroclimerization of APJ and KOR. and reduced ERKl/2.. t1-ie lefi ventricle. Apelin. in both doses reducecl the level of heterodimerization in 2K1C animals,::td recoverec'l the reduction in pERKI/2. Inhibition ol KORs significantll,increased ERKI/2ion in response to apelin 60pg/kg, Chelerythrine ii-rhibited apelin 40 and 60- indLrced60- induced increase in ERKI/2in [:RK1/2 phosphorllation. I']'l'X inhibited apeliniation.Conclutions: Findings showed that the OPR. particr-rlarly KOR, mediate the inotropic...itropic. ar.rd depressor eflects of- apelin. 'fhe interaction of the OPR and APJ augments the-.,-,tropic and r,'asodcpressor cfl'ccts of apelin. 2K1C hy,pertension increased the expression of:.ORs and heterodin-rerization of APJ and KORs. 'l'he heterodirnerization r.l'as associated by'.-.luctiorr of ERK phosphorl'lation and alterecl the cardiac inotropic and lusitropic efl'ects o1',, relin. I'hese changes ma)'par"ticipate in pathopliysiologl,of cardiac dysfunction in renovascuiar.',pertension that is irssociatecl u'ith subnorn-al lcvcl ol'scrlur apelin. Apelin- induced recov-er!", ERKl/2 phosphory'lation and KOR-APJ dirnerization may nominate apelin as a therapeutic--,rl ir-r trcatnrent o1-this kincl of h.vpertension.

بررسی نقش گیرنده های اپیوئیدی کاپا و تعامل آنها با گیرنده های اپلین در اثرات اینوتروپیکی مثبت و منفی اپلین در قلب موش صحرایی نر مبتلا به پرفشاری خون کلیوی مزمن

http://eprints.kmu.ac.ir/26795/2/6314.pdf

بررسی نقش گیرنده های اپیوئیدی کاپا و تعامل آنها با گیرنده های اپلین در اثرات اینوتروپیکی مثبت و منفی اپلین در قلب موش صحرایی نر مبتلا به پرفشاری خون کلیوی مزمن

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