Shahouzehi, Beydolah

Simorgh Research Repository

,tt{11P,t,,5,; '', 5 r(,1,'.13|.J Y'.',,1Jti .-rll--a# 4i,;j J,-,-sJ .S-rS" lt"- *ti ,!!sREBP-2r SIRT1:.llJ..,AMPK .rt*+ * microRNA-33,-., ..-,,51*,1 s ,-, . ,.*,3S I ol;31"Fo. *5 ;.r AMPK-SIRTI Jis JJ-,oJJ-aL:! oll":: ' L, ,.i6J-os,-o .r-!*)li ;t:S.l :t-rat3 .tt: "'lz-)s ,',-t,-,.. ;iS.l :;11,k- lti*,l\_ v$\ Y11- \ Y1V :.Jf,--: Jt*,Kerman University of Medical SciencesFacultv of Medicine!In Parlial Fulfillment of the Requirements for the Degree PhD of ClinicalBiochemistryTitle:Effects of microRNA-33- agonist and - antagonist on AMPK, SIRT1 &SREBP-2 expression in AMPK-SIRTI pathway in mice liverBy:Beydolah ShahouzehiSupervisor:Dr. Gholamabbas MohammadiAdvisor:Dr. Hossein FallahYear: 2017-2018:,,ftfev /.- "L-.-osr5 .:;-1 cf +L,o ! l#) ,--1,,, i x* 9 S )" ;L-l &tr e. ),,-l G9f .# .Shf.rL* :.!lsl 9 as*iLjL;rrA,r) cs.{r.-Eb "6 (SREBPs) Jst-1.5 or:ll ,,L;; ,-;-c JL;i dt6 ,-yqgy-9 (LXRs) .grJX ".<-:l, (RCT) Js*# r-pf*Jbl i G.ft pe o.:1.: llp rb c,, l) SREBP .!t! L LXR .$5- s)\ e.*i ,.*JlLl, ,-J.;' ,r ;t- .F; +f -u" o "rl5 rt)+ 9 s|, ;9ss *f dlaRNA lo microRNA .-r:"r* o*-;,l .rr;"" CPTl , ABCAl ,AMPK .Jkcj ,fio ,bK :.- miR-YY .-u5,," ,s)\ Jl- 6lx.5G a*b ,,-*.r-V S aJlt ,r .ro: ,l;i )fib'c^d' l) SREBP , LXR ;Jti ;-l 2:G ;y-)L.l-I.: .-l-P ;l SIRTI ,F-, ir-,SIRTI ,AMPK ,_,Q Sy l, miR-YY .5 o-r5 ,kr !. 9 miR-YY mimic ! ol,r* LXR *;9fi ,4 y .-.e)f ,s,.ty S;y, cyr -r"51: ABCAl 9 SREBPs ,LXR-o'.o9F )y,,r,, V ,,L*; n! , (*.f * ;s &r 2) s-r"; ea,*; ,9f 2 ss tssyu Gr r, F s s- idgt,lmgikg/48h nriR-33 mirnic u os.F,(T0901317)LXR .4fi -30 nrg/kg/-l8tr er o9r5 ,;rlsl;l .5h; -(e ,miR-33mirnic e LXR ;*;ef\ ,.4 y e, "e.S.s:r5 cjLr.: 1 mg/kg/48h mR-33 inhibitor ,l* "g,S :;la- $l: ,-y.si Jty-t 6,,1"r d; dab .s:rf ;.sL-r: miR-33 6 o.s;S )i{d , LXR *lfi -J; ,t;; a".:s\$ smiR-33 l!,;1.,*" .+.:f sD\ e.+ j5 ci! I er" 4y* 9 o$ oJ-,-;'.,;r -,U19o )g;Y )l rn.ri-r":; pu>.-,.y,.gx ;lre ,-1*; g ,4).f ;**-'Real-timePCR ,ys:\Abcal, AMPK, Lxr-a, Sirt1, Srebf2, Srebfl.x rr" .5tb{*-J .s r$l .<+yi or-,9 ,yss s,.!,,,9 ABCAl : AMPK ,LXR-u, ,SIRT1 ,SREBP-2 .5;.$-rj J).5 olrsl .r-;i;\*:,, 1l o:b-.. -(r ':.5) es g1 .^S r; g (Y ,g.5) miR-33 qr o-r5;t4f ,F ,s.f) T0901317 aJ :l: JL; L ryb :b c!t,,,) .!-),5 ,fl ,-i^5 c.cL .g;l: G* :F. (Y .i,5)miR-33mimic 9 -u pr- HDL-c &.lPl9*, .irl, G* ;y=)l{d 9 LXR c-;951?..5i tl r; oroL; SREBP-2 9 miR-33 .r!.l.pl ,:r5 c-.sL-r: LXR c*;r5i a5 Y -....;Ll r"i AMPK .r[, JoK .-*- ;*q..o T0901317 .:l: J.bti l) miR-33 e SREBP-2 ;L miR-33 .i .-.:9,r r1.: .r* ,,-* --Jo! .r; AMPK.T! ;oi5 s &.lfl *yr,e.,1u.Erl; .r;* ,* *. miR-33 r'nimic a,*;3.^o 9 miR-33 .r-* *- LXR Ji-: JLr of r-:-bre JL; t .5Lo c.iL- .rls ;L; .911r .r; i)4il a,Jlt :,y Sl^ og.f ,: SIR,.sl:rK-*r, IrLXR-u oQ ss: T0901317.s os_lpl ;1 miR-33 rkr I SABCA1 gLXR-o. j1.3 _;.jor5llq 9dr.1pl l,SREBP-1c;t*g;,b G*;hmiR-33n.rimic!olr*'10901317,*S;9T090i-:I-Alr s)t ,r* thmiR-33 mimic ! "l:* T09013 17 ?S ;.1.:l: ,;o15 l) SREBP-1c .r) of, nT iR-3-: ..s;9.;,1:,r.;- olrn+;l ,+Jl.t :,,y,,sQa:.5;: J9;:*J5 x1n 1osr.l".l:l:.r;l;-il l; ey'+;-{ _; -Ql-rt * 9,1 ,.(J9l.;,-Ekr-,,L- 9 ;9[<*l 9.;1.glr, Lo9,,l:.-cJ r: o-r"; pb.,l cjkpF*","rr"1" :6y,! a>u;i,s-r"" il,cl 11 .5r^9 .Jl{r+ ar )9r-JS Jful i o:l: ioiS1; ,)}- }b J9;-l 6rt- ;;-z-o g +.,*J : -.rF )p rb-r- slp ;if ,-*b .Skr_,,t"* 9 ;9JS*lryi ).ry, )) .r-F.E ;L;r ;l9r t t: U_l ,i y t; _-.-4fi rt .1^.t ,.fi ) or"j ;*-r, "5 o-r52l9r:,1 e_li o, a+i ! I S rl2.;l S..jly.* L_Ir.,, y)1.> 6 u:.r)'**.r,..r+L-),-(;9rI(-19"1 .tli*JlaJ-r.1:11 9:u-l .rti*;g"c:1;l;19-""miR-3i)l+d"L-'.'..:lr,1;i a,Jlt ,:19..-sF ;. *LXR-u (SREBP (AN,IPK crliR-33,LXR---*fl,;elS-le;\ :6.4L< .. JAbstract:Background and objectives: Cardiovascr,rlar diseases (CIVD) are stil1 the leading cause of-ntortalitv and morbidity' n,orldu'icle. Liver X receptors (t.XRs) and sterol regulatorl' elementhinding proteins (SREBPs) pla,v an irnportant role in lipicl homeostasis. LXRs al'fect SRf,BPs andincrease lipicl sl,nthesis and reverse choiesterol transport (RCl'). MicroRri:\s are endogenous" non-coding. short RNAs r.vhich play pivotal roles in regulating n:retabolism aud other aspects of cellbiolog.v. MicroRNA-33 down regulates AMPK, ABCIAl and CPI'1 mRNA. On the other hand,SIRTl afI-ects LXII and SREBP activity through deacet,vlation. In this stuch'r,re evalr,tated effectof co-administlation o1'miR-33 r.nirnic. inhibitor and LXR activator on AMPK-SIR]'1. LXR-u.SREBPs and ABCIAi expression iu mice liver.Materials and methocls: 'l'hirty six mice r.vere divided into six groups (n:6) and the study duratir.rnu,as 7 da1,.s. Group I received standard chow without any treatment. group II received 30 mg/kg/;t8hLXR agonist (1'09013i7). group lll received I rng/kg/4Sh "LNA mil{-33 mimic". group IVreceir,,ed 1 m-u/lig/48h ''in vivo LNA anti-miR-33". g1'olrp V received both T0c)01317 and "LNAn'riR-33 rlimic'". ancl group VI received both 1'090i317 and "in vivo l-NA anti-miR-33". Alltreatments rvere aclnrinistratecl througli i.p iniection. After 7 da,vs, uice u'eLe sacritlced and livertissues u,ere excised and blood samples were collected. miR-33, Abcal" AN{I'}K. Lxr- o. Sil't1.Sreb12 anil Srebfic gene expression w'ere quantified by real tirne PCR. and ABCA1" AN'IPK. l-XR-u. SIRI'I ancl SREBP-2 protein expression by u'esteru blotting. Sertitn lipicl profiie also wasmeasurecl. Serunr lipicl protile \\.as assa)'ed b1' enzvmatic rnethods.Results: Our lestrlts showed that T0901317, ar.rti-rniR-3i ancl co-aclntinistration of'T'0901317 andanti-miR-33 significantll, increased t'IDL-c levels and miR-33 rnimic significarrtll reduced it.'f0901i17 increasecl SREBP-2 ancl miR-33 levels but co-adtninistration of'T0901317 and anti-miR-jireclucecl srebD and miR-33 levels. AMPK u,as reduced b1' T0c)tJ 13 1 7 insignificantl-v. Anti-ntil{-33 11c1 r.r-riR-i3 ntimic signiticarr-rtly increased and decreased ANII']K expressiot.t respectivell'.Sir.t1 gene ancl protein erpression rernained unchanged in all six grollps. Our resr-tlts showed tiratI-XR activation canse LXR-g and ABCIAl eler.'ation and miR-33 inhibition attenrtates T0901317etfect on LXR-cr explession. '10901317 and conrbination o1'T0901317 and miR-33 rnin-ricsigniticantll, increased and anti-miR-33 reduced SREBP-1c gene expressiot.t. '10901317 andcornbination of T0901317 and rniR-33 min'ric significantly'it.rcreased'i'C levels.i rircussion: In spite o1'progress in drug discovery for atherosclerosis and tnetabolic diseases, these- :,.rrders are not fully controlled. It seem that there is a s-vnergistic efl-ect betu'een T0901317 and'.i-nriR-33 rvhich reduce serlrlrr TG lerrels. and also reduce intracellular sterol levels and iucrease- .,.',iesterol transport to hepatocl,tes, theretbre this can be considered as a therapetttic alternative in-rerosclerosis and cardiovascular diseases.F,er rvords: Atherosclerosis. LXI{ agonist, miR-33- AMPK. 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Effects of microRNA-33- agonist and - antagonist on AMPK, SIRT1 & SREBP-2 expression in AMPK- SIRT1 pathway in mice liver

http://eprints.kmu.ac.ir/26893/1/6375.pdf

Effects of microRNA-33- agonist and - antagonist on AMPK, SIRT1 & SREBP-2 expression in AMPK- SIRT1 pathway in mice liver

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