4
DOI 10.13350 j.cjpb.190602 ·!"· miR133a3 456789:;<=>?@ LX2 !A α SMA BCDE op 2qr stu ,vv wx yz{|}·~ 1. ¯ü:B#Bü&°e³(C$ùúÑÅ}[DEF{78ö¿GuF=HIF3=DEJKLM 830054 2. DE78<N{8³ÈÉ8=DE<ËFde"ÞúÑÅ}[3. DEF{78dOF8=<8³A*<8Pû[!"#$ QB miR133a3p úfRS&Trecombinantadenoassociatedvirus rAAV vÍUVazW-XY Zop¬T W-JÝ'~l%& a[XYZopi LX2ops}Af DMEM QfAAV8Scram bledQfAAV8miR133a3p fl qRTPCR PVW miR133a3p W - Í U UVW \ H ;<COL1A1 )、 ] ^^_;<alphasmoothmuscleactin α SMA @ TGF β SmadÁÂà`deZ Í U U'( ú f R S & T ° aQBzW - LX2 opAAV8miR133a3p W - LX2 op f Z miR133a3p ¶Q Í U Uf 11000±343.10 ³ DMEM QfËÇý8bc =32.07 0.01 )。 AAV8miR133a3p W - f ³ DMEM f α SMAmRNA Z¶QÍUUAjf0.42±0.06vs1.00±0.09 =5.72 0.05 ), COL1A1mRNA Z¶Q Í U Uf 0.34±0.03vs 1.00±0.04 =12.66 0.01 )。 ') úfRS&T°aW- LX2opï*XYZop$ α SMA ZÍUfd miR133a3p e)XYZop.è#Z~lfT8g!dO*+,RS&TXYZopmiR133a3p -./01R373.9 23456278116735234 2019 06062504 犑狅狌狉狀犪犾狅 犘犪狋犺狅 犲狀犅犻狅犾狅 犵狔.2019Jun 14 ): 625-628. 犆狅狀狊狋狉狌犮狋犻狅狀狅犳犃犃犞8狋犺犪狋狅狏犲狉犲狓狆狉犲狊狊犲狊犿犻犚133犪3狆犪狀犱犻狋狊犻狀犺犻犫犻狋狅狉狔犲犳犳犲犮狋狅狀α 犛犕犃犲狓狆狉犲狊狊犻狅狀犻狀 犔犡2犮犲犾犾狊 LILiang ZHANGChuanshan BIXiaojuan MA Yangyang WANGHui TUERGANAILI Aji LINRenyong 犛狋犪狋犲犓犲 犔犪犫狅狉犪狋狅狉 犘犪狋犺狅 犲狀犲狊犻狊 犘狉犲狏犲狀狋犻狅狀犪狀犱犜狉犲犪狋犿犲狀狋狅 犎犻 犺犾 犘狉犲狏犪 犾犲狀狋犇犻狊犲犪狊犲狊犻狀犆犲狀狋狉犪犾犃狊犻犪 犆犾犻狀犻犮犪犾犕犲犱犻犮犪犾犚犲狊犲犪狉犮犺犐狀狊狋犻狋狌狋犲 犜犺犲犉犻狉狊狋犎狅狊 犻狋犪犾犃 犳犳 犻犾犻犪狋犲犱狑犻狋犺犡犻狀 犻犪狀 犕犲犱犻 犮犪犾犝狀犻狏犲狉狊犻狋 犝狉狌犿 犆犺犻狀犪830054 犡犻狀 犻犪狀 犓犲 犔犪犫狅狉犪狋狅狉 犅犻狅犾狅 犻犮犪犾犚犲狊狅狌狉犮犲狊犪狀犱犌犲狀犲狋犻犮犈狀 犻狀犲犲狉犻狀 犆狅犾犾犲 犲狅 犔犻 犲犛犮犻犲狀犮犲犪狀犱犜犲犮犺狀狅犾狅 犵狔 犡犻狀 犻犪狀 犝狀犻狏犲狉狊犻狋 犇犲 犪狉狋犿犲狀狋狅 犅犻狅犮犺犲犿犻狊狋狉 犆狅犾犾犲 犲狅 犅犪狊犻犮犕犲犱 犻犮犻狀犲 犡犻狀 犻犪狀 犕犲犱犻犮犪犾犝狀犻狏犲狉狊犻狋 犃犫狊狋狉犪犮狋 犗犫犼犲犮狋犻狏犲狊 Toconstructan AAV vectorthatoverexpresses miR133a3pandtoexamineitsinhibitory effecton α SMAmRNAexpressionintheLX2hepaticstellatecellline. 犕犲狋犺狅犱狊 LX2cellswereculturedinvitro di videdintothreegroups andtransfected withthreeagents DMEMcontrol AAV8ScrambledgroupandAAV8miR 133a3pgroup .TherelativeexpressionofmiR133a3pandtheexpressionof α SMA COL1A1 caspase9 andTGF β SmadmRNAweredetectedusingqRTPCR. 犚犲狊狌犾狋狊 AAV8miR133a3pwassuccessfullyconstructedandtransfected intoLX2cells.miR133a3pwasoverexpressedintheAAV8miR133a3pgroup.Therelativelevelofexpressionof α SMA mRNAinLX2cellstransfectedwithAAV8miR133a3pwas0.42±0.06.Theleveldifferedsignificantlyfrom thatintheDMEM controlgroup =5.72 0.05 .TherelativelevelofexpressionofCOL1A1mRNAwas0.34± 0.03.TheleveldifferedsignificantlyfromthatintheDMEMcontrolgroup =12.66 0.01 .Therewerenosignifi cantdifferencesinexpressionofTGF β SmadmRNA 0.05 犆狅狀犮犾狌狊犻狅狀 AAV8miR133a3pwassuccessfully constructedandtransfectedintotheLX2cellline.TheresultsofinhibitionofoverexpressionofmiR133a3pindicated thatAAV8miR133a3pisanoveltoolforfurtherresearchonliverfibrosisregulatedby HSCandmiR133a3p. 犓犲狔狑狅狉犱狊 Adenoassociatedviruses AAVs ); hepaticstellatecell HSC ); microRNA133a3p · · 5 6789:;<= 犑狅狌狉狀犪犾狅 犘犪狋犺狅 犲狀犅犻狅犾狅 犵狔 2019ôõ ö14÷öø Jun.2019 Vol.14 No.6 9:;#DEhi`C6`j{8dpÿÒNo.2016D01C329 ); `j{8dpÿÒNo.81371838 81660108 ); DE hi`C6kD0ldmÿÒNo.201705120 )。 <=>?nopEmail renyong _ lin sina.com >?@Aý q1983- ), r5s7012Å}tòóû3O4XuÜv³w÷ZA**Email lilisx83 163.com

miR 133a 3p456789:;?@ LX 2!A SMA BCDE · .1 MN •†‡…—–ƒ⁄pq*rsxy犔犡2K\ 犉犻犵.1 犚犲犮狅犿犫犻狀犪狀狋犪犱犲狀狅狉犲犾犪狋犲犱狏犻狉狌狊犲狊狑犲狉犲狋狉犪狀狊犳犲犮狋犲犱犔犡2犮犲犾犾狊狅狀犿犻犮狉狅狊犮狅狆犲

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Page 1: miR 133a 3p456789:;?@ LX 2!A SMA BCDE · .1 MN •†‡…—–ƒ⁄pq*rsxy犔犡2K\ 犉犻犵.1 犚犲犮狅犿犫犻狀犪狀狋犪犱犲狀狅狉犲犾犪狋犲犱狏犻狉狌狊犲狊狑犲狉犲狋狉犪狀狊犳犲犮狋犲犱犔犡2犮犲犾犾狊狅狀犿犻犮狉狅狊犮狅狆犲

DOI:10.13350/j.cjpb.190602 ·!"

·

miR133a3p456789:;<=>?@

LX2!AαSMABCDE

op

1,2qr

1,stu

1,,vv

1,2,wx

1,yz{|}

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1,3

(1.¯ü:B#�Bü&°e³(C$ùúÑÅ}[

,DEF{78ö¿GuF=HIF8û3=

,DEJKLM830054;

2.DE78<N{8³ÈÉ8=

,DE<�ËFde"ÞúÑÅ}[

;3.DEF{78dOF8=<��8³A*<�8Pû[

【!"

】 #$ QBmiR133a3púfR¶S&T

(recombinantadenoassociatedvirus,rAAV)vÍUVazW-XY

Zop

,¬T W-JÝ'~l

。 %& a[��XYZopiLX2op

,s}AfDMEMQ�f

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bledQ�f

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,�lqRTPCRPVW miR133a3pW-ÍUU

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(COL1A1)、�]

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(alphasmoothmuscleactin,αSMA)@TGFβ/SmadÁÂà`deZÍUU

。 '( úfR¶S&T°

aQBzW- LX2op

,AAV8miR133a3pW- LX2opfZ miR133a3p¶QÍUUf11000±343.10,³

DMEMQ�f����ËÇý8bc

(狋=32.07,犘<0.01)。AAV8miR133a3pW-f³DMEM fαSMAmRNA

Z¶QÍUUAjf0.42±0.06vs1.00±0.09(狋=5.72,犘<0.05),COL1A1mRNAZ¶QÍUUf0.34±0.03vs

1.00±0.04(狋=12.66,犘<0.01)。 ') úfR¶S&T°aW-LX2op

,ï@ø*XYZop����$�α

SMAZÍU

,f�dmiR133a3p�e)XYZop.��#Z~lfT8g!dO

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】 R¶S&T

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;miR133a3p

【-./01

】 R373.9    【23456

】 A    【2781

】 16735234(2019)06062504

[犑狅狌狉狀犪犾狅犳犘犪狋犺狅犵犲狀犅犻狅犾狅犵狔.2019Jun;14(6):625-628.]

犆狅狀狊狋狉狌犮狋犻狅狀狅犳犃犃犞8狋犺犪狋狅狏犲狉犲狓狆狉犲狊狊犲狊犿犻犚133犪3狆犪狀犱犻狋狊犻狀犺犻犫犻狋狅狉狔犲犳犳犲犮狋狅狀α犛犕犃犲狓狆狉犲狊狊犻狅狀犻狀

犔犡2犮犲犾犾狊

LILiang1,ZHANGChuanshan1,BIXiaojuan

1,MAYangyang1,2,WANGHui1,TUERGANAILI

Aji1,LINRenyong

1,3 (1.犛狋犪狋犲犓犲狔犔犪犫狅狉犪狋狅狉狔狅犳犘犪狋犺狅犵犲狀犲狊犻狊,犘狉犲狏犲狀狋犻狅狀犪狀犱犜狉犲犪狋犿犲狀狋狅犳犎犻犵犺犾狔犘狉犲狏犪

犾犲狀狋犇犻狊犲犪狊犲狊犻狀犆犲狀狋狉犪犾犃狊犻犪,犆犾犻狀犻犮犪犾犕犲犱犻犮犪犾犚犲狊犲犪狉犮犺犐狀狊狋犻狋狌狋犲,犜犺犲犉犻狉狊狋犎狅狊狆犻狋犪犾犃犳犳犻犾犻犪狋犲犱狑犻狋犺犡犻狀犼犻犪狀犵犕犲犱犻

犮犪犾犝狀犻狏犲狉狊犻狋狔,犝狉狌犿狇犻,犆犺犻狀犪830054;2.犡犻狀犼犻犪狀犵犓犲狔犔犪犫狅狉犪狋狅狉狔狅犳犅犻狅犾狅犵犻犮犪犾犚犲狊狅狌狉犮犲狊犪狀犱犌犲狀犲狋犻犮犈狀犵犻狀犲犲狉犻狀犵,

犆狅犾犾犲犵犲狅犳犔犻犳犲犛犮犻犲狀犮犲犪狀犱犜犲犮犺狀狅犾狅犵狔,犡犻狀犼犻犪狀犵犝狀犻狏犲狉狊犻狋狔;3.犇犲狆犪狉狋犿犲狀狋狅犳犅犻狅犮犺犲犿犻狊狋狉狔,犆狅犾犾犲犵犲狅犳犅犪狊犻犮犕犲犱

犻犮犻狀犲,犡犻狀犼犻犪狀犵犕犲犱犻犮犪犾犝狀犻狏犲狉狊犻狋狔)

【犃犫狊狋狉犪犮狋】 犗犫犼犲犮狋犻狏犲狊 ToconstructanAAVvectorthatoverexpressesmiR133a3pandtoexamineitsinhibitory

effectonαSMAmRNAexpressionintheLX2hepaticstellatecellline. 犕犲狋犺狅犱狊 LX2cellswereculturedinvitro,di

videdintothreegroups,andtransfectedwiththreeagents(DMEMcontrol,AAV8ScrambledgroupandAAV8miR

133a3pgroup).TherelativeexpressionofmiR133a3pandtheexpressionofαSMA,COL1A1,caspase9,andTGFβ/

SmadmRNAweredetectedusingqRTPCR. 犚犲狊狌犾狋狊 AAV8miR133a3pwassuccessfullyconstructedandtransfected

intoLX2cells.miR133a3pwasoverexpressedintheAAV8miR133a3pgroup.Therelativelevelofexpressionofα

SMAmRNAinLX2cellstransfectedwithAAV8miR133a3pwas0.42±0.06.Theleveldifferedsignificantlyfrom

thatintheDMEMcontrolgroup(狋=5.72,犘<0.05).TherelativelevelofexpressionofCOL1A1mRNAwas0.34±

0.03.TheleveldifferedsignificantlyfromthatintheDMEMcontrolgroup(狋=12.66,犘<0.01).Therewerenosignifi

cantdifferencesinexpressionofTGFβ/SmadmRNA (犘>0.05). 犆狅狀犮犾狌狊犻狅狀 AAV8miR133a3pwassuccessfully

constructedandtransfectedintotheLX2cellline.TheresultsofinhibitionofoverexpressionofmiR133a3pindicated

thatAAV8miR133a3pisanoveltoolforfurtherresearchonliverfibrosisregulatedbyHSCandmiR133a3p.

【犓犲狔狑狅狉犱狊】 Adenoassociatedviruses(AAVs);hepaticstellatecell(HSC);microRNA133a3p

·526·

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Jun.2019, Vol.14,No.6

【9:;#

】 DE�hi`C6`j{8dpÿÒ

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Jun.2019, Vol.14,No.6

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Jun.2019, Vol.14,No.6

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[1] PinzaniM.Pathophysiologyofliverfibrosis[J].DigDis,2015

(33):492-7.

[2] KitanoM,BloomstonPM.Hepaticstellatecellsandmicrornasin

pathogenesisofliverfibrosis[J].JClinMed,2016,5(3):E38.

[3] FengY,NiuLL,WeiW,etal.AfeedbackcircuitbetweenmiR

133andtheERK1/2pathwayinvolvinganexquisitemechanism

forregulatingmyoblastproliferationanddifferentiation[J].Cell

DeathDis,2013,4(11):e934.

[4] PucheJE,SaimanY,FriedmanSL,etal.Hepaticstellatecells

andliverfibrosis[J].ComprPhysiol,2013,3(4):1473-92.

[5] ArriazuE,RuizdeGalarretaM,CuberoFJ,etal.Extracellular

matrixandliverdisease[J].AntioxidRedoxSignal.2014,21

(7):1078-97.

[6] YoshidaK,MurataM,YamaguchiT,etal.TGFβ/Smadsigna

lingduringhepaticfibrocarcinogenesis[J].IntJOncol,2014,45

(4):1363-71.

[7] GordonKJ,BlobeGC.Roleoftransforminggrowthfactorbeta

superfamilysignalingpathwaysinhumandisease[J].Biochim

BiophysActa,2008,1782(4):197-228.

[8] ð;)

,ñòó

,óÎ.W�<:e*βQXYZop½�Ze

)~l

[J].HIXò&9$

,2016,32(11):2195-98.

[9] SanjayS,GirishC.RoleofmiRNAanditspotentialasanoveldi

agnosticbiomarkerindruginducedliverinjury[J].EurJClin

Pharmacol,2017,73(4):399-407.

[10] DuanLJ,QiJ,KongXJ,etal.MiR133modulatesTGFβ1in

ducedbladdersmoothmusclecellhypertrophicandfibroticre

sponse:implicationforaroleofmicroRNAinbladderwallremod

elingcausedbybladderoutletobstruction[J].CellSignal,2015,

27(2):215-27.

[11] WangL,LiX,ZhouY,etal.DownregulationofmiR133via

MAPK/ERKsignalingpathwayinvolvedinnicotineinducedcar

diomyocyteapoptosis[J].NaunynSchmiedebergsArchPharma

col,2014,387(2):197-206.

[12] RoderburgC,LueddeM,VargasCardenasD,etal.miR133a

mediatesTGFβdependentderepressionofcollagensynthesisin

hepaticstellatecellsduringliverfibrosis[J].JHepatol,2013,58

(4):736-42.

[13] OhnoK,ItoM,KawakamiY,etal.CollagenQisakeyplayer

fordevelopingrationaltherapyforcongenitalmyastheniaandfor

dissectingthemechanismsofantiMuSKmyastheniagravis[J].J

MolNeurosci,2014,53(3):359-61.

【hijk

】 20190305 【lmjk

】 20190510

·826·

5 6 7 8 9 : ; < =

犑狅狌狉狀犪犾狅犳犘犪狋犺狅犵犲狀犅犻狅犾狅犵狔 2019ô6õ ö14÷ö6ø

Jun.2019, Vol.14,No.6