Diazoxide opens the mitochondrial permeability transition pore and alters Ca... Hideki Katoh Nobuhiro Nishigaki:Hideharu Hayashi Carewlanon:Jun 4,2002,105,ProQuest Medical Library Pg2666 Diazoxide Opens the Mitochondrial Permeability Transition Pore and Alters Ca2+Transients in Rat Ventricular Myocytes Hideki Katoh,MD,PhD:Nobuhiro Nishigaki,PhD:Hideharu Hayashi,MD.PhD Backgroumd-The mitochondrial Kam channel (mitoK)has been implicaed is an end effector or trigger of ischemic precoaditioning (IP)Although a mitoK.opener.diazoxide,mimics IP.mechanisms for the cardioproceetive action remain unclear. Mfethods and Rerir-We measured Ca transienes (CaTs)and mitochondrial inner membrane potential (with confocal microscopy and the flucrescent probes fluo-4 and tctramethylrhodamine ethyl ester perchlorate in rat ventricular myocytes.Diazoide inereased the amplitudes and diastolie levels of CaTs dose dependently.The effects of diazoxide on CnTs were inhibited by the mitoK antagonist sodium 5-hydroxydecunoic acid (100 umoVL),whereas application of diazoxide cad e change inAfer sarplasmic rium fnction was disabkd with ryaodine and thapsigargin,the efeets of dioxide on CaTs were stinl observed.The opening of the mitochondrial permeability transition pore was monitored with flucrescent calcein.Diazoxide icoelerated the lakige of cakein from mitochondrial matrix (16%of control:Pc005),and this effect was inhibited by eycksparin A (2 umolL).Cyclosporin A also sbolished the effeets of diazoride on CaTs.Dinzoxide nxidized flavoprocein flucrescence reversibly,and this eftect was partially blueted by cycksporin A (by 24;P.c0.0)5). Conelurions-We conchade that in rat vencricular myocytes,diazoxide modulues the opeeing of the mitochondrial pemeability transitice pore.relting in increuse in CaTs independent of the cbanges inThe action of diaroide on the mitochondrial pemeability transitiom pore also affeets the mitochondrial redos state.(Ciresfarion.2002:105: 2666-2671- K行Wand:myocyles■ichemia■ealcium [schemic peeconditioning (IP)is n phercmenon in which trantion pore (mPTP).causes permeabilization of the mito- Ltramient nonkethal periodk of iscsemia increase the resis chandrial memhrane and prvides ancher pathway for Ca tance to a whsequent proknged isehrmic period.Various efflux.Cyclosporin A (CsA)is kaown as a specific ishibitor sbstances and signaling pathways coad he involved in IP. of mPTP.Classically,irreversible opening of mPTP has been ineluding sacolem ad mixchordrial ATP.sensitive po. implicned as an early event in lethal cell damage (ie, tassium channels (sarcoKn and K Emerging evi- apoplosis).2 Rccent studies.however.sggesed that dence has suggested the contnbution of minoK.as an end mPTP could contrihule to Ca homeastasis under physiolog- effector or trigger of [P.*Althcugh it has heen demomrnted ical conditions.4 that diaroxide.a selective mitoK opener.mimicked the In this stucty,we dimned to里extigale the changes im△t effects of IP and that sodim 5-hydruxydecancic acid(5-HD) and Ca transients (CaTs]by diacmide is intact myocytes a mitoK channel inhibixr,abolihed the pemtective action and showed that duzoxide modalaled mPTP and inereased of IP and diaroxide.the precise mechsaisms for the Ca"trunsients via a CsA-seasitnve pathway. cardioprotective action of diazoxide remain elusive.It has been speculated thathe opening of mitoK dissipanes the Methods mitochondrial ianer membrane potential (leading to a reduction in mitochondrial Ca"uptake. Cells and Solutions Milochondria play pivomal roles in the maimenance of at vestricul myys were prepared as described peeviousy. cellular Ca"hemeestais.*Ca'uptrke cccurs throagh the r3 cyoes were plaoe过i量3 chanher and super向sed with stand时 Ca uniporter drives by The mechinisms for Cad Tymode soluion.A xrimonts were condected第2CSa时 efflux are Na'/Ca'exchange (NCX)ind Ca/H'ex- Tymde souror contained (mmolL)140 NuCl,6 KCI,I MgCl:.S HEJES,5.8 tluoe.and I Cl:(pH 7.4 wih NaOH)Cels were change.'The opeming of the proteinaceous pore in the fild semalated 1 Ha)with 2ms wohape pubes of 1.5 times the miochondrial membrane,ie.the mitochondrial permeabilty hresto对amplo Receivod2l.202:eb4n小tM线a2i4 cepted Mareh22 From the Depurtrrert of Iaketsal Medicise III.Hanimelsu University School of Modie ng,Hrmimata,Japat. Commpondence t Hideki Kh,MD.PtD.Deperrent of leml Modicine IIl.Hananass Uriverty Schoed re Medicine.1-20-1 Handa-yama. Hamanaie.Japan 431-3192.E-mal hcthehana-edazjp C 2002 American Hean Assocstin.Ire. Cireadation is availle httpewwwxirolonahearg D0:R1i5U1.GR6N3L4目64级44 2866 Reproduced with permission of the copyright owner.Further reproduction prohibited without pemmission
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Diazoxide opens the mitochondrial permeability transition pore and alters Ca ... Hideki Katoh; Nobuhiro Nishigaki; Hideharu Hayashi Circulation; Jun 4, 2002; 105, 22; ProQuest Medical Library pg. 2666
Katnh et al Diazoxide,mPTP,and [Ca'] 266万 B 2 160 12 0 500 10 0 wO control diazoxide 80 10 um 60 0.4 40 20 wnton-teaamb 0 200m5 ctl Dz Rigure 1.EFocts of damoxide on Ca?transerts and reoting spark tecusncy.A.Cels were exposod to dlaxoxide (200 pmol]and feld at 1 H2.CaTs were recordod st 10 minutos tor af.B.Doos-dependonl efocts of an CaTa.Ditcgcod物was apped位Mtively te sc,ti3i,Dala repnsent rticn±9Ee的4 aaperment,C念ameed ie sin im cont inonsity,whom F:indcates booal fuoresconce;cd,camol;Dz,dinoxide and we,woahcut.'Pe0.05 va control (poirod t toof Measurement of Ca'Ts and Ca+Sparks (escitation at 458 nm and esiicion al 505 to 550 ami were cecoeded. CeTs were leded wit 20 umalL flao 4-AM for 20 iniues ond floressert inersity or oe regint of inerest was inegruted. Flaovesserce imaging vas pertommed oith laser scaining cortocal mcos:3 pe ILSCM:Ce,Norni c0pd1o3 IInvemo对miroscop网 Measurement of Flavoprotein FTuorescence Axiovent S100,Zelss)wih a 63x waler immersin objoove chordial rdoo sre wa insesaod by meiurtmel国lee Inumerical apertare [NAI1.3 Zeiss1.excitation waelength of 488 oeace of FAD-lisked enzsmes.'Eaddsenous fvusroxin fuore om.and emossion ar 5 am cere25e633dd4eic6NG如h24阅nrd Image aoqumition foor quantitatise analysis of CiTs was mde in n0nMy5055》1 hwne-n mode (scanned at50e小以n时CTem上id mTse释n网h3akeg徐r3 line In h2 Stntisticnl Analyses meagimnt of Car carks.was caluisinl,ind viaually Resolrs are expressed5 inean上SEM tor the Indi:edr的erC iseatified Ca“Mrt作pled if lueal Cchanges excerdod nyocytes from at leos 3 amimals Semstical sigificanee was 价nmo以with duratien a-lilade>s由s” detemmined by paire间dI好分er ANOVA,Values of Pctins w ×起l义cl. Meurement of Membrane Putential in Mirochondri To rieestre v cells were loaded wih H nmoVL omathyl- Results rhodamine edhyl eser perchkrane (TMREI for 20 rinules.Images of Diazoxide Increases CaTs and Ca2+Sparks wrre ohtiiard IMHI is dictnhued hetween alir bumpu:liunb Figure1Ad在mm1s位at pertusio0CNai2ode2l ording lo Nems's ruation.5 For the meruremenl of tihe- gmolL)fur 10 minules inencased diastolie kvel and the dependem TMRE forecenl chatpe imuees were secteded tvery umplinudes of CaT's.Duse-deperdent effects of diaxouide an 30oewd(ilh向alin time为s43smg:.F月ue:时a CNTs are:sueriz in Figre IR.whid:shows that inared uvr resims of inkeresl (60xGO pixes placed over the concentrations of dizcxide 200 umolL alleral CaTs. brigh porios of ys Thes suhsaquent experiments were condocted with 200 Imaging of mPTP Opening With Calecin umolL dsazoxide.unkss ochereise stated.Diazoxide also Cells were lroaded with I umoliL cal:rin-AM f mirus aid the increased resting C子·spaurk fregpoency frum379+l28 queaching of cyoonlic caceis was achieved by addition of sfuk5~飞，'，s05数9±138paks·pL-1·s'P005: 5 remoll.CoC m the sulation.Hages of calein fluorescenoe n-5:Figures IC and ID). Reproduced with permission of the copyright owner.Further reproduction prohibied without permission
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2666 Cireulenon June 4,2002 A 25 10 Figure 3.SR dd not contribue to efects al diazodde.A.Appl- functic0=abalshid by 5 an广gLh年srargin and 5 am0L Figure 2.Dazoxide incresed S Cn-ponlent bu did nat sler Diastelo leves al F./Fe wore compared.FF:nd catas norma saten ol fud stiullcn cooing 110 mmol s applod ctl co-trol:De cl2Dd8rd%0,w*Gn0w情rprb网st rpldy to eveke CafTs.B.Pecled data af implituce of CafTa mean=SEM.'P0.05 vs cortrol (paired testl jas index of SA Ca"cortonl.C.Pooled data of timme constant 0.uthr用Tmea士5 EM Tom H b:erimar.G1ia decttased the TMRE sigrsal remxrkably (th am nsults comtro;Dz,d azoxide:and wo,washo.t 'Pe0.06 vs comrol were cbserved int 3 other cells)We next investigated whether 闲red t te. S-HD could inhihit the eftect or diazoxlde cn CaT's.In control experimears,reapphcation of dia江o次2 er the tirst expo Next,we imvestiganed whether SR Ca coetent and the sure and washo caed the same increase in CaTs as the sctivity of srcolemmal NCX were affected hy diazoxide nitial applc流iana属sha4.When cells were ireate Afrer feld wimuletion was omed,caffeine (10 mmolL? wich S-HD (106 pmol/L)for 10 minutes befoce the seccad wis applod rapidly (Figure 2A)ind the amplitude of pplcaoa subseqent application of xide in the pres. eatfeine transienes (CafT's)was measured as an estimabe af enoe of 5.HD did zot increase CaTs (Figures 4B and 4CX SR Ca coment Figuee 2B demenstrated that diazoxide These results indiested that although opening of mitoK. inend the amplinude of CafTs slightly but significanthy (by 5.00.2 versus oontrol:P005:n-8).The deelining eould te imlvod depolarization of was not correlated with the observed increase in CaTs ta突of CalTs溶fitted o single exponenial decay tn index C etfux via NCX.There were ro effects of dianoie on CsA-Sensitive Puthwuy and CuTs Ca efflux vis NCX.beeause the lime constarts of the CT A peevious study with ixolaled mitochondria showed thar decline were not changed by diarouide (Figure 2C) diacoxide released Ca?fm mitochcadriz via a CsA. To address whether incressed CaTa pould he attributed tn eisitive mechuism Thus,we eximined whether CsA the direet etreets of dinzacide on SR.we appled diaxoide uhibits the effects of diazoxide on CaTs.After expcewure and ader conditions such that SR functon was inhibited Cells wene pretreaed with ranodine (5 urc)and thapsigargin w2女cut of dia配nxk,cee样rwih24 nolL CsA fur I5 minules Reapplicutikm of dizoxide with CsA did not 13 pmolL)foc 15 minuks.and then disreuxide was applial. ineresse CaTs,which indicates that 2 CsA-sensitivo mecha- The effect of diazoxide on CaTs was not atteruated under these conditions (Figures 3A and 3B),which indicales that nism ws responsible for the increase in CaTs (Figures 5A the effects uf iixcoxide on CaTs were not medinced d rectly and 3B).Becanse CsA is a potem inhibiter of mPTP.we via SR investigsed the effeets of disroxide o mPTP by measaring calein signal index fer mPTP apening.Perfusion of Diazoxide Does Not Alter Mitochondrial Tymce solution for 10 minetes decreased the calecin signal Membranc Potential ty63年.and perfusion of Tyrode pis CsA〔2umaL It has been speculated thi the operting of mitek wonkd caused only a slight decrerse in signal imemsiry (by 1.9 dissipoe edg to decreased diving focre for the Ca dsra not shown)Diazoxide decreased the cakem signal by uniporer.>The reduction of miloctril Ca''apeake onuld 1(005),and there was litk chamg:when diaroxide inerease eyxsolie Ca".Thus,we manitored the thanges in was applicd with CA (20 Figure 5C).These results A.before and after application of diazoide.As shown in sugpested that there was a CsA-sensitive leakage c cakein Figire4A,diazouide30 molL）did not alver△t ro minochodria and女at diaroxid女Jcceleraled this leak- Application cf FCCP(2 uo)after washot nt diazouide age sigraficantly. Reproduced wth permission of the copyright owner.Further reproduction prohibited without permission
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Katoh el al Diapoxide,mPTP,and [Co2l 2669 3120 w▣ 40 10 20 30 Tme (min) 2 11 Figure 5.CaA innbita efocta of cinzooode an Ca trareders.A hera3pta水n3ndw"0tg时d2dsce6 were incubaled wth Co (2 umovL)for 10 minutes.Subacquont aopicadon of cla zactie with DA ed no irererc CiTa B.Podl:daln from 4 s0wme80ut悟oprestrt mean±S日，Pc0C58coto pairod r too.C.Time co.rsos of charpes in ca pe n sipnal during perfireo al coto Tyror:e solulen nd during perfrion cl 0南有十mpWg4Cb周nd atmsence ij of GsA2um n-4 -0.6 vs diamooda plus CeA and tPeC.05 vs Tyrode's ANCVAL F/Fa incicas rormal mod fuoresoant iclensty,whe gure 4.Dx正M4目fa成ler itcchon士mmnoranD Fo indcates basa fuoreseroe:cf.ertmt and Cz.cazmeice 5.HD.A Ropresertat va rocordinp of TMRE sgnal after applca- Dinzoxide und CaTs tien of d 500 mo ad FCCP 2 HmolL)TMFE sig 3lWwN5eape5E丹对35务ct片r洋hanty口n炉8+me9 Here.we showed that daxoxide increased CaT's dase depen. dently an:l thaf 2200 gumnoli.was raquireal.whidhn was clnse cHb wens Iraet:l wth 5-HD:100 prtalI lor 10 mnunes, to the repered concentration runge of diaouide for cardio. nd daz3e5p3edh5-HD.GP0dt4 s.F/F.indicatas nommaizec nucrescem niensity.whare F: A Da市年阳5e-tm1nr5EM.'P边05得G3Tp3ru对号10 ■R■ CsA-Sensitive Puthway and Fluvuprulein Oxidatiou To investigale the poessible Finis between diazoxide and the mitocboddrial metabolic xtale,we measured flavceroein funrescelce to index the mitochondrial redox stak.The 【doxa知dw5 cahbrated b的cxpusune nf the<el,o 1004010 2.4-DNP (100 uo/L)fellewed by sodium eyanide Time Intin (2 mmal As xown in Figure f,diazoxide reversibby B iml山ed exidaon of flavopente to37%of the D公NP value. and CsA 12 gmolL)partially atenuated the ouidative effect of diazexide by 24 IP005.n=4:Figune 681. Discussion In thix study.we demonstrated that diazoxide incrrased Cal's.Cafls.and Ca'spark frequeney withoul using signif2 n changes in Ad,aidC%A aneuated the e士ect图 DECsA of diasoxide on CaTs.Diazoxide caised flavaprotein Figure 8.Cnidrtve ef色ofdi必s4 taly inhibtec Ey oxidocion.which was partialy inhibited by CA.These CAA P amale4AA0a道e20cfq@f寸0mp5ntb results indicate the possible invulvement of CsA- voproen sg Signd wax nomaleu:l as 100%far 2,4-DNF D0 xmol/L-induoed moamum coddaton ad for scdum cr sensitive mechanism for the effeets of dinzoxide on cyto 0%2 mmo L)-inducoc8 mplate ro山ction.日.co可do元m solie Cu'homenstasis aund the mitoctocdrial redox state. d9pw阳wmnn=8E1.P005aC-Id11以. Reproduced with permissian of the:copyright awner.Further reproduction prohibited without pemmission
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2670 Cireulation June 4,2012 procection (10 100 AmnclL).However,a higher concen. ociginated from mochondria we compred the fluoreserr Iration of diazuxide could huave unexpecle loxic eferts en inensilies before and afier sapenin permeabalization of the mitochondrial function as a metabolkc inhibiter and a proe- sarcolemma and foud that reducti of the calcein signul nnaphore.x We have observed that even al 11)wtol/L uI保，We lave als hscrvedl th减inee diseuxide hs efTecls inxin myocytes (upuhlishal dalak decreased when cells were colaed with TMRE,in agree. Therefore.the reduced acoessibility of diazoxide to mito- menl with an criginal repet fdals not shownl.Tasen chondria in inmact myocynes might account for the relatively together,these resules indicale it is likely that the cakein higher concentration.An inereasen Ca]by ditoside has sicnal msaly originated from milochondia.We showed that been repored previously. diazoxide accelered the calcein leakie from mitochondrin, Several madanisn enukl be imobed in the inensse in and fhs effcls wen:altemelol by CsA.which sugpests the CaT's.soch as (1)icressed Ca entry via srcolemmal Ca possible oction of dazoxide an mPIP. channels.(2)decreased Ca'etfux via NCX and Capmp In uur expecrimental conditions.CA alone did noc aer and (3)increased Car nkiee from SR.Here.we dmon- CaTs (dira not shown).and CsA per se had few effects o SR stned that diazoxide did noc alter Ca'efflux via NCX and NCX in r myocyles34 Thus.cur results that CsA (Figure 2C)In m mce-tes,SR dmmnalex for the repulatxoe inhibined the eect of diazoxide Cs support the th cf Ca',during twitcb.ind unly small portions af Cur'were mPTP mighl contribute to the increase in CaT's. ex:nuded via the Ca?parp.a It has been shown that The opening ot里PIP bas heet nociuted wit造dixsipalinn diaexi:e ducs nul afed 1 Tenefone,neither the clangex of a,nualrix swelling.and unkuupling of oxidalive phee- in Ca''indlux or Ca'"efflux via sarcolemma conld coctribuie phorylation and plays a key role m apopois by releasing to the increased CaTs.In additie there was nn direct actioe ylochrome&,ai书ogniced th满in ischemi风perfu- of dazoxide on SR (Figire 3)Thus.diazoxis does no sion injury.some cells undergo zpoptotic cell deach us likely modulae theseChandling sysiems.Wang et oppod to rs and that mPTP may el as the trigger for preh渊blockade of CaTs b的enpommil in diide apoptasis.In this regard closing but not openang the mPI'P is peetreated hearts abalished cardiac protecticn and sugpested iteal ta prevanl ll daath.This migl he:the cx:in the porsihle inwolvement of sarcolemmal Ca chornek for ireversible opening of the mPIF.Dizxoide was reported to activation of mitoK.hy diazoxide.Becau we showed that inhihit the relas of tochrumecid prevened aoplosis in Ca"release from SR wus anlikely to be involved as a source ofelevated Ca,the cnpes in (as a tigger af Ca.induced myocyies esposed to hydrogen perotde.However.isalated mitechendria have been repored release cytechrume e in Ca release)hy diaoxide might not alter CaTs sigrficantly. resparse to K openers. The different experigental conditions (myncytes or Fvi h:n revealal the etixene:af the firkering o Largendorff-perfial tears and the exience o'ischeria mPTP (lew coaductance stane.A brief and reversibe dng protcols)may eaplain these diffeen nPTP opesing does not cause mitcchondrial swelling,is not Diazoxide and Mitocbondrial Membrane Potential relaned to the release of cytochrome c.and might serve as a Previous reportsed thal opening mitoKdepolarized phyxiokical means of ridg mitecbondria of excess me. 3ik ind reducel mitoeondraal Co"uptaice.'>Ocr resuls. tabolnes oc is.in particlr Ca Our results indicae thl however.did sol sppunt [his hypohissis,hess hen:wen: diaenil .ould indlace a loww-cnndigunce state of mPTP mid no detsctable changes in by diaroxice.There ane releaseCafroe mitochoadria.whichfferen phen xrssihk esplanations for undeteced changes in First, enon fmom irreversible mPTP opening.Frevious studies the K influs through mitoKw could be compensaled rapidly demonstrated that a mPTP-celated Ca'nelease oecumed in cugh that we could not follow'the changes i证△t.刘 coeditoes in which mitochondria were relatively Ca''over. Secund.depolarizalion of d was too small in be dexecsed looded.However.it is not likely tha mitochondrial C hy TMKE.Lawrence ct ald reperted that similc cunvatr- concentraticns C were hemvihy looded in the present tions of diazoxide as used a this study (100 to 200 umoVLl 女y.Although C.'门a palhways with lw-lu.aeg did pot thanp:bul proteeted myocytes from memabolkc mPI'P remain urresolved.both mPTP per se and the revenod inhibition.Another study.hooever.demonstrated thi cardio- mode uf the uniporter migh be involved. proective effects or dinzoxide during ischemizreperfusion Investiganon od [CaL is essemial to elcidate the action have been associated with mild depolarizaron ef△g.力n of diazoxide on mitochondri.Hawever.[Ca"'L measure- the experments with isalaed moochondria and neonatal ment with confocal microscopy and a Ca indicakor (eg. diae myucyle.,liah全deo山ri2d3kbn15mV. Rhsl-2:in inlact myseylex has not heen eahlished. whereas Kowaltowski ct als reporial thal eimted chag The mechanism for the Jdke of dixceuxide un mPTP is or Ad from calculated K"influx through miKwere only poxcly uncersnocd.I has been snggested that loo I to 2 mV,Differem experimental conditicns and different conductance mPTP i3 principaly openned by ma编rap明 eiho达ould accom以foc these diserep维ci felevnced pH opens mPTP)rather than Ca".In reltion lu this,it hs been postulated that cpering of mitokcaes Diazoxide and mPI'P Opening K"influs,and this charge could be compensated for by The kodling of rll with enkein-AM plus cobalt to assess proton catrusicn,leuling In raisel manix pH.which tnagers mPTP opening was inilially nportal i hejneytes hat nce low conductance mPTP.Addibocal mies an:neassury l in cardac myocytss.To confirm that the calecin signal clarify the precise mechanisms ivclved. 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Katoh et al Diuxuxide,mPTP,and [Ca 267】 Regardless od the duration ct opening.the mPTP opening mirst be acoompemied by the depularization of Ad.However. Thha:1LEee山”nrhh销B4长 1大1烂：4-7 Petronilli et 2 repored thaut a short PTP apening was detected only hy the calcein-plus-cohalt method wherecs 4A3.195.124:139-16. TMRM stribution required longer mPTP openis Anther :and ire mda p元blem is血e production of re5 ave oxygen spocies.uhicb eId2t,n:w到人32-N are poteer mPTP.inducing agents,by exposure in the capublt of prtmig ird comeyitg clednall ard cicum aigtah.CnN. lr.Thus.海e shorkened laser excicali0ndia,江dhe 1k145-1153 intervals for each recarding were proloaged to 30 secends. The relegsed Ca?"fmom mitochondria could tall the SR and dinc d hydropyrilie ieeeptor角wdre peiryor：videxce fum 3wlCM.2208206-1. pnevide sufTicicnt chanpes uf (C in a liny localial It.icp1,n上wC,是ashrtxr L ci.rranarehendrt Cr and to ictivane ryanodine receptcrs.Thes could explain the in- creased SR Ca"conents and Ca"spork frequency ificr the nAv f Phro.H网75月44H9 addition of dazoxide Ichas et al#suggested the inkeraction cTIK话t:rElocandral pernea力均ndta0p元cnw between mitochondria and SR within restricled spoce as the tard dirertly nrtact cels by change is ritrehcadril cakein fo- mechanism tenned mitochordriz Ca-induced Ca"release. Kawuluiki Nl.Seedurann 5.Putek P.tl al Biveseredtic oune- .of te AlP-acttnc K'u her MITI and Mitochondrial Redox State A时/nL201:2134号-185， We demoastrated that diaxoxide increased flavoprutein uxi- darie and that CsA pamally inhibied this effect of dizxoxlde d aboAxie fnxd prokction it olikd m Furts.Am PAai. 2300wh25-H255. (Figures 6A ad 6B)Thus,although the exae process is 20.nwa Jw.Ipuaai IA,Bers DV.Ralxctkn :n rathk and n card a: unknown.the effects of diazoxide on mPTP could be relabed elk spedlesceprnkers dimein%相lg事aTK/Ph Is the mexechondrial redax stope.Hecouse [Ca"L stimulztes 19044617-25. key eneyms for NADH mulin the reiluepd 1l.e灯T名xi灯Lt3工om by wtxhi Tik上au1 may irbibit NADH oxidation.Thus.the diseuxik-infuoxd 22.Lawverce CL Billges B.Rodrige GC.et al.The K.chonel opene low-coeductance mPIP corl serve ts an endogenous vncou- dk家 pler of oxidative phospharylatim G2rgT利十na PA1w4.20:34535-542. sumeary.we provide evidnce that diazouile mudifies te lawonductance stat af mPI'P and that there are eroklmry n Girai edib ud C2C12 mps indsce pussible linkx betwer mPTP,ICL,and the mitochandnal hn上aln34in.m.0块- redux sate in intact myocylas.Audilional sulies ane rojuired 2AsMR4,lrt相LmoC文d.d6 porin ithtia日B to elucidae the underlying mechanisms beiween tte AT了hT.I9处26是H19-日I70 diazoxide-isduced mPTP opening and cardioprotective attioas. 约yrsk与l起1c1e51n图Tmkc线o poiit an odharv.rAi Repes Ar,1999:136:177-19 2AM.h灯A.O'Rcerke B.&..AT:ewI References ot对un九rreb int女pehL站Ur格u道6 tcLy uf llal cdll injry i isarig rpoeman wat Iw 27.Bulnstaned EL.Jeanmic S nrej FP nN.Minchondtal ATT- 7k1114-1136. 】tenY.lrc%,ayM.【emc pacontoring:fro W以8i25H15约-HIS6 enie代r0Kw6加I形A型R物2月企S-0 2线.cta下，A5 IP.Tren cakian型Talrg划2 I datt:twe 】.aH材D),Paucek P.Yes-YuADy V,el aL Condi4ieci国 daagtkds and itx interaaitt wrh mtctordrial ATP.eskline K' m归-初Th(ordectnce swle01tdM5A15L355 结如 102-10g2. 2移gtsd4发4，Ciad:TL Permeahel5 ntlare porc u the i wr？Twm新Anrd52以.I月3tI7-lbl. :si5.i6mg的8 w.1g62.13-6 S.Lu Y.Suo T.O'Rnrke B.AT7-cepexan: aoun cfanne在Jdc51E0aGft10了C 17T2463-246级. 31.Sheu SS.Shet VK.A novel ieghniqse for qaanristo nawreroe af 长WugY,HU图K.Ashp M Actiation of mhochredrial A-sir K~t拉t2ga0s,归na图ie injury k depedat on 57754 C:uctvity.Cin:Pri 1999.15:731-741 好1，a星.1Thag T.iale Kn,Tacck P Yam-Ynmuy V.c al T:uildokil K ulL6∞of cnochroe#e3 nd caT dke3 h omelaies wah址 of per:in si.J CAo 201:73-1217-12TH 1935271809%-55. 及DB.nh.口度K地L0,Ravts (R05卧 hesr程z:arw phenark1 n xtemuaying iducti006f时 cpanin prvea Ca"overlaad in nl casdic mikdhondkia.J Psl 155551937-30 2c32:l0m1-l04. 头.Dch2 R.Comributio of niochand由o aniral prpe妇r fmer t行otitic AcTE经oltn si1lgA制6cGch/hIg mgabrin ne rurrnain irtrarikxhrhal It:xbolim Phys Pe 411-17. 101591-425 10 Gmfer TE.Gamer KK.Sheo 55 al Hiachrndrtal cuctmn teppart seind Etbtax4 emial ond i扣，0CE8m 21C以 Repraduced with permissinn of the:copyright owner.Further reproduction prohibited without permission
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission