細胞凋亡的線粒體途徑

細胞凋亡的線粒體途徑蔣舜媛董霞程在全2002-12-17報告提綱

細胞凋亡的特征細胞死亡損傷性死亡

Necrosis程序化死亡、細胞凋亡ProgrammedCellDeathApoptosis細胞凋亡的特征形態(tài)學特征:

染色質(zhì)的凝集,嗜堿性染色增強,細胞核崩解此時線粒體保持形態(tài)正常.細胞體積縮小,一部分細胞質(zhì)和核碎片進入由膜包被的程序死亡小體,他們從細胞表面出芽脫落,并被巨噬細胞.上皮細胞吞噬.生化特征:

染色質(zhì)降解,核小體間連接DNA部位被降解,產(chǎn)生寡聚核小體DNA片段,即180-200DP整數(shù)倍的不同長度的DNA片斷.Fig.1.Schematicsummaryofbiochemicalmechanismsofapoptosis.MitochondriaandCommitmenttoCellDeath線粒體是真核細胞的重要細胞器，是動物細胞生成ATP的主要地點。線粒體基質(zhì)的三羧酸循環(huán)酶系通過底物脫氫氧化生成NADH。NADH通過線粒體內(nèi)膜呼吸鏈氧化。與此同時，導致跨膜質(zhì)子移位形成跨膜質(zhì)子梯度和/或跨膜電位。線粒體內(nèi)膜上的ATP合成酶利用跨膜質(zhì)子梯度能量合成ATP。合成的ATP通過線粒體內(nèi)膜ADP/ATP載體與細胞質(zhì)中ADP交換進入細胞質(zhì)，參與細胞的各種需能過程。MitochondriaandCommitmenttoCellDeaththeeffectorsofapoptosisarerepresentedbyafamilyofintracellularcysteineproteasesknownascaspases.Inhibitingcaspases,however,doesnotalwaysinhibitcelldeathinducedbyproapoptoticstimuli.Althoughcaspaseinhibitorsblocksomeoralloftheapoptoticmorphologyinducedbygrowthfactorwithdrawal,etoposide,actinomycinD,ultraviolet(UV)radiation,staurosporine,enforcedc-Mycexpression,orglucocorticoids,theydonotnecessarilymaintainreplicativeorclonogenicpotential;ultimately,thecellsdiedespiteinactivationofcaspasesbywayofaslower,nonapoptoticcelldeath(6-9).Incontrast,antiapoptoticproteinssuchasBcl-2,Bcl-xL,andoncogenicAblcanmaintainsurvivalandclonogenicityinthefaceofthesetreatments.Conversely,someproapoptoticproteinssuchasBax,amammaliancelldeathproteinthattargetsmitochondrialmembranes,caninducemitochondrialdamageandcelldeathevenwhencaspasesareinactivated(10).Suchexperimentalobservationsarguethatacaspase-independentmechanismforcommitmenttodeathexists.Thismechanismislikelytoinvolvemitochondria,aswewillsee.MitochondrialPathwaysinphysiologicalcelldeath

thereleaseofcaspaseactivators(suchascytochromec),changesinelectrontransport,lossofmitochondrialtransmembranepotential,alteredcellularoxidation-reduction,participationofpro-andantiapoptoticBcl-2familyproteins.MitochondrialPathwaysinphysiologicalcelldeath

Ifmitochondriaarepivotalincontrollingcelllifeanddeath,thenhowdotheseorganelleskill?Atleastthreegeneralmechanismsareknown,andtheireffectsmaybeinterrelated,including(i)disruptionofelectrontransport,oxidativephosphorylation,andadenosinetriphosphate(ATP)production;(ii)releaseofproteinsthattriggeractivationofcaspasefamilyproteases;and(iii)alterationofcellularreduction-oxidation(redox)potentialDisruptionofelectrontransportandenergymetabolism

disruptionofelectrontransporthasbeenrecognizedasanearlyfeatureofcelldeath.

γ-Irradiationinducesapoptosisinthymocytesandadisruptionintheelectrontransportchain,probablyatthecytochromeb-c1/cytochromec(cytoc)step.Ceramide(a"secondmessenger"implicatedinapoptosissignaling)disruptselectrontransportatthesamestepincellsaswellasinisolatedmitochondria.LigationofFasalsoleadstoadisruptionincytocfunctioninelectrontransport.DisruptionofelectrontransportandenergymetabolismOneconsequenceofthelossofelectrontransportshouldbeadropinATPproduction.Althoughsuchadrophasbeenobservedduringapoptosis,itoftenoccursrelativelylateintheprocess(14).Indeed,ATPappearstoberequiredfordownstreameventsinapoptosis(15).Thus,althoughlossofmitochondrialATPproductioncankillacell,itisunlikelythatthisisamechanismforinductionofapoptosis.Disruptionofelectrontransportandenergymetabolism線粒體跨膜電位的耗散與細胞凋亡有密切關(guān)系近年來陸續(xù)有報道說明線粒體跨膜電位的耗散早于核酸酶的激活，也早于磷酯酰絲氨酸暴露于細胞表面。而一旦線粒體跨膜電位耗散，細胞就會進入不可逆的凋亡過程。線粒體解聯(lián)的呼吸鏈會產(chǎn)生大量活性氧，氧化線粒體內(nèi)膜上的心磷脂。實驗證明，用解偶聯(lián)劑mClCCP會導致淋巴細胞凋亡。而如果能穩(wěn)定線粒體跨膜電位就能防止細胞凋亡。Releaseofcaspase-activatingproteinsTheimportanceofmitochondriainapoptosiswassuggestedbystudieswithacell-freesysteminwhichspontaneous,Bcl-2-inhibitablenuclearcondensationandDNAfragmentationwerefoundtobedependentonthepresenceofmitochondria(16).Subsequently,studiesinanothercell-freesystemshowedthatinductionofcaspaseactivationbyadditionofdeoxyadenosinetriphosphatedependedonthepresenceofcytocreleasedfrommitochondriaduringextractpreparation(17).Duringapoptosis(invitroandinvivo)cytocisreleasedfrommitochondriaandthisisinhibitedbythepresenceofBcl-2ontheseorganelles(18,19).Cytosoliccytocformsanessentialpartofthevertebrate"apoptosome,"whichiscomposedofcytoc,Apaf-1,andprocaspase-9(20).Theresultisactivationofcaspase-9,whichthenprocessesandactivatesothercaspasestoorchestratethebiochemicalexecutionofcells.Releaseofcaspase-activatingproteinsSignificantly,caspaseinhibitorsdonotpreventcytocreleaseinducedbyseveralapoptogenicagents,includingUVirradiation,staurosporine,andoverexpressionofBax(14,21,22).AnexceptioniscytocreleasefrommitochondriainducedbythetumornecrosisfactorreceptorfamilymemberFas,inwhichcytocreleaseispreventedbyinhibitionofcaspases(primarilycaspase-8)recruitedtothecytosolicdomainofligatedFas(21).Nevertheless,cytocreleasecansometimescontributetoFas-mediatedapoptosisbyamplifyingtheeffectsofcaspase-8onactivationofdownstreamcaspases(23).Theemergentviewisthatoncecytocisreleased,thiscommitsthecelltodiebyeitherarapidapoptoticmechanisminvolvingApaf-1-mediatedcaspaseactivationoraslowernecroticprocessduetocollapseofelectrontransport,whichoccurswhencytocisdepletedfrommitochondria,resultinginavarietyofdeleterioussequelaeincludinggenerationofoxygenfreeradicalsanddecreasedproductionofATP

Reactiveoxygenspeciesandcellularredox.Mitochondriaarethemajorsourceofsuperoxideanionproductionincells.Duringtransferofelectronstomolecularoxygen,anestimated1

to5%ofelectronsintherespiratorychainlosetheirway,mostparticipatinginformationofO2.Anythingthatdecreasesthecouplingefficiencyofelectronchaintransportcanthereforeincreaseproductionofsuperoxides.Re然ac慎ti盼ve寒o川xy熱ge桐n縮慧sp懇ec崗ie訴s詞an腦d悄ce仰ll維ul資arre場do垃x.Su險pe爺ro漫xi巡壽de胃san糧d橫li怖pi礙dpe買ro熟xi帆da軋ti和onar雙e笛in飾cr性ea承se秀ddu彎ri傾ng邁a柴po恢pt級os晚is滔i漸nd活uc汁ed切b泰y譜my斤ri楚ad釘s獻ti典mu獅li慰(28).Ho圓we慢ve婚r,鵝g爭en定er短at派io隆n睬ofRO肅Sma叫y薯be葉a趕r遭el墻at洗iv物el娛y腐la殃te銅e陽ve介nt容,糊oc宰cu貿(mào)rr勇in幻玉g歸af釋te斬r象ce頑ll肝s嘗ha宿ve逐e莊mb尿ar戲ke胸d硬on亂a鄙p混ro像ce清ss何o則fca甚sp昂as警eac徒ti吊va恭ti毀on劍.In午t災hi孔s未re槳ga丹rd量,舉at蠻te催mp通ts嘩t覺o肆st趙ud耐y旬a(chǎn)p幸op禾to談si犬s寫un久de躬r儉co交nd大it筆io銀ns帖o惠f話an娃ox根ia宅h歲av珠e由de熊mo游ns累tr艱at帽ed純t庫ha抖t豬at泉l旗ea左st蓄s裁om哥epr荷oa儀po溝pt千ot穿icst捆im觸ul樹i織fu顫nc僻ti倆on攔i鞠n展th扇e岡ab削se豬nc倦e秋or拉n誼ea胸r泛ab仁se桿nc罵e員of太o詳xy版ge絲式n,烈w垃hi浩ch終i投mp炊li防es儉t路ha買tRO軌Ssar屑e擺no四t袍th遇e翅si右ne眼q岡ua資n揀on旁o酒f閱ap堂op奇to押si差s趣(29,30).Ho榜we吩ve太r,RO撓Ssca然n珠be猛g栗en決er醫(yī)at孝ed執(zhí)u使nd盡er言c瘡on狀di頌ti其on賓s贏of筐v沿ir露tu劈燕alan信ae奶ro草bi撤os覆is(31),淋a值nd纏t銜hu機s股th懷ei廚r戶ro程le坐i近n巧ap賺op瞧to劃si覽s稿ca原nn洗ot挪b簽e礙ex吼cl寒ud省ed壘s躺ol鎮(zhèn)el新y晨on符t掃hi沉s串ba蹲si獨s.Fi劃gu坦re雷2摔.Mo徹de顏l丈fo元rca蒜sp務as爆eac衰ti稍va盛ti泛on梁b欺y系mi頁to狠ch港on仙dr雅ia悼.os講mo答ti昏c增di襯se雹qu哈il海ib演ri超um驅(qū)l噸ea拌di鐘ng傍t用o畏an渠e諒xp藏an籌si禿on咳o瓣f皇th塞e阿ma爛tr個ix桶s彎pa袍ce擦,or絮ga抓ne牧ll壩arsw午el教li擾ng紀,堪an捉d急su淘bs齊eq也ue廊nt牌r茶up偵tu導re廁o剪f拜th劍e鵝ou藥te程r秧me丘mb班ra嶄neth貢e販ot蒼he榆r鞠en氧vi泊si殺on途s境op培en奸in處g叔of困c塵ha憐nn柄el眾s版in匹t柿he鏟o異ut間er封m迎em數(shù)br燃an順e努th襯us醫(yī)r壓el郵ea遍si拾ngcy敵toc趣fr姨om錘t矩hein跑te躍rm吵em嘩br段an肢esp蝕ac沒e宋of潮m拘it注oc叼ho積nd找ri泛a幸in蛇to貌t開hecy鏈to殿so般l.PT軟P型or丸eIn發(fā)m飾an刮y狼ap登op啊to稀si刺s拋sc怪en口ar脂io戚s,令t役he臟m值it舞oc籠ho姑nd和ri廉a(chǎn)l緩i繩nn繁ertr投an宮sm賊em旅br溉an育epo陽te針nt遙ia條l閃(m看)族co姐ll足ap買se狐s材(32),悟i損nd筍ic咱at絲式in文g碎th抓e破op陽en惹in盟g朵of戀a沸l(wèi)匙ar爸ge閃c糟on系du緒ct妙an字ce閣c間ha題nn劫el個k應no早wn字a種s路th情e賭mi紙to乎ch遺on提dr現(xiàn)ia盆lPT跌p植or草e(33)駁(Fi極g.企2).it翁s風co姑ns賞ti盼tu虧en燦ts梯i虎nc看lu字de閃b槐ot愉hin曉ne濁rme到mb躍ra巷ne庫p駁ro揪te波in改s,鮮s效uc載h披as林t云head印en秀in擾e牽nu早cl庸eo繭ti淡detr論an反sl泡oc辣at偵or(A歌NT乏),an瀉dou饒te蜘rme蹈mb蠅ra愿ne士p萍ro森te頌in犬s,粗s膏uc收h滑aspo架ri對n(vo喂lt蘆ag暮e-膏de貌pe更nd工en典t團an寸io駐n站ch勢an溉ne甜l;疼V肯DA脹C)攜,wh餃ic犬h咐op女er值at涂e既in啄c連on孩ce療rt睬,戴pr既es恨um昆ab飲ly故a背t菌in襖ne半r侵an珠d第ou娃te庸r尋me滴mb遭ra擔ne忠c蹈on隙ta行ct叔s抗it脊es血,嗓an糾d腿cr賓ea爬te喊a湊c遲ha線nn殿el訂t訪hr叫ou酬gh食w孟hi治ch叛m敞ol遲ec未ul歌es≤1.膨5kDpa棒ss刷(32,34).PT辦P隙or蜜eOp耕en駐in泊g纏of蠢t低hi褲s班no嚼ns軍el攔ec幸ti惡ve連c格ha享nn匪el注i滔n贈th匙e僑in翻ne靠r帝me翅mb襲ra咽ne丘a膽ll間ow向s逃fo豬r煩an弦e酒qu漆il翠ib勿ra珍ti板on這o豪f鎮(zhèn)io支ns針w如it奴hi呈n況th鋒e魂ma慢tr過ix宅a挎ndin奏te隙rm我em吧br協(xié)an劉esp腹ac視e軟of惡m再it紐奉oc血ho慨nd制ri唉a,彈t悟hu磚sdi郵ss恩ip軌at合in壺g閑th繼e妖H+洋g們ra泥di綢en損t葡ac建ro點ss旗t狗he閑i揉nn拾er吹m杰em椒br性an瓶e脹an夕d笨un絹co杜up翅li恐ng伸t狠he計r拉es傭pi俗ra縮慧to編ry汽c定ha蒸in.Pe摔rh椒ap蜓s專mo煙re貍i代mp渠or偽ta殖nt縱ly丑,貼PT菌p鋪or傍e遣op肌en如in徒g踩re寄su換lt各s傘in攔a泛v粒ol歪um言edy湯sr傻eg嗽ul單at象io騙nof敗m株it柴oc資ho單nd佛ri席a過du碗e事to匯t銀hehy逢pe矛ro在sm史ol襯al繳it聾yof飛t歉he解m毛at仰ri兄x,薯w移hi壘ch愉c演au微se脈sth間e撒ma猛tr絞ix艇s望pa且ce懂t蠢o拼ex鄰pa尖nd.Be誘ca順us同e士th狼e辜in愚ne鈔r假me腸mb掌ra堡ne效w落it向h蘆it代s段fo鉛ld緞edcr泳is埋ta盆epo拳ss宜es作se謠s射a濁la肥rg各er農(nóng)s裕ur雖fa濫ce孟a牛re失a助th事an文t窗he腸o衛(wèi)ut楊er沖m茄em壩br酒an蓮e,灶t庸hi暮s筍ma波tr詢ix碑v嘩ol鴿um哥e縱ex膏pa昨ns將io碗n妹ca患n膜ev剝en史tu悉al脖ly鹽c艇au悶seou垃te賊r攪me姜mb生ra卷ne務r誘up刪tu閉re,re忌le兆as鉛in平gca懂sp速as扛e-a飄ct錢iv鞋at雅in繳g餡pr典ot怪ei昌ns專l嗓oc領(lǐng)at頁ed仁w白it五hi會n勤th屠ein灰te們rm蒙em藏br見an卵esp蕩ac繡e澤in涉to夢t鄉(xiāng)豐hecy弦to柴so活l(Fi辱g.注1).Fi徒g.袍3楚.Th欲e糾mi鑰to楚ch筒on奴dr避ia前l(fā)呈pe疏rm耳ea婚bi羽li驅(qū)ty纏t朋ra梨ns吧it畢io紫n.PT掩P扔or澇e的性序質(zhì)通過祥一些健實驗怎室的越研究果，以倍下諸嶄點值井得指奸出：線粒周體內(nèi)近膜通旱透性屯轉(zhuǎn)變牽既是名細胞酬凋亡筍的必譽須條哈件，派也是肅它的液充足栽條件際。PT孔道妖打開姐后導自致線麗粒體簽許多肅功能拌的致趨命性即變化蠶從而著啟動狠了死敬亡途癥徑。PT孔道冊作為芬許多鳳生理剩效應贊的感者受器沿（二膏價陽贈離子征、AT罵P、AD奏P、NA塞D、ΔΨ擇m、pH、巰切基與鳴多肽)，整抖合了龍電生楚理、菊氧化爺還原布與細那胞代塑謝狀賀態(tài)的桂信息千。PT孔道退的組關(guān)成成秧分AD多P-婚AT努P載體峽是能咳量代翅謝的揉重要階分子盼，由母于AD歷P-埋AT消P載體襲是由顏一個較基因柱家族戚的幾西個成塵員所凱編碼精，它直的表猾達有驅(qū)嚴格瞎的組括織專凝一性濫。因焦此，PT孔道歡在不徹同細梳胞中娛的調(diào)茂節(jié)可彈能稍撿有不役同。PT孔道蜘的作色用有籌自放曉大的睛效應告。PT誘導ΔΨ棵m耗散烤，而掠反過減來mC艱lC名CP使ΔΨ例m去極座化會煉導致PT。一淺些PT的結(jié)仇果例恰如ΔΨ羞m耗散償，活幅性氧顏的生丸成本岸身也壇會導亞致PT。這線就說植明PT會有閃正反誕饋，代從宇而在派細胞未凋亡視中有厭自摧謝毀的日作用證。反噴過來察，如胸果能億防止ΔΨ夾m的耗漆散，儉就能盞避免辦氧化伏還原多不平腳衡、這磷酯該酰絲活氨酸督的暴凱露與訂蛋白差酶和粗核酸番酶的制激活朽。PT奴P悟or編e有開歌放與攜關(guān)閉從二種撿構(gòu)象PT孔道昨開放蒙導致鐮細胞抵凋亡績。在PT孔道跡開放牧時線剃粒體典釋放蝕細胞命凋亡米誘導框因子(A休IF浮)。AI逆F可能浮是一散種蛋壓白水諒解酶包，位沉于線露粒體滲膜間怪間隙呢，它崇能被旗蛋白陷酶抑占制劑著如N-芐氧階羰基-纈氨國酰-丙氨疏酰-門冬另氨酰政氟甲姥基酮(N妖-be壺nz蜻yl豎ox裹yc敬ar青bo粉ny鄉(xiāng)豐l-增Va墻l-椅Al喇a(chǎn)-糞As森p-甘fl辣uo且ro鄰me外th隊yl導ke計to柜ne)所抑栗制。蒼術(shù)堤苷促確進PT通道障開放和。蒼術(shù)敞苷只愉能與AD煎P-再AT笑P載體算的胞恢液側(cè)剝結(jié)合而。此外連從線嘗粒體騎釋放繭的細賊胞色鹿素C也是嘩一種圍細胞依凋亡斧誘導鼓因子梯。而PT孔道朵關(guān)閉國能防特止細糟胞凋度亡。當PT孔道昆與環(huán)皇孢菌的素A(冊cy磁cl承os睬po奶ri付nA)或SH，或厲米酵局菌酸(bo攝ng質(zhì)kr烤ekac下id遇)結(jié)合烏時PT孔道呼被關(guān)救閉。米酵穿菌酸丘可與AD擇P-臟AT膀P載體激的胞畜液及主基質(zhì)勒二側(cè)構(gòu)結(jié)合Fi潔gu就re憑2販.Th好e涂mi門to慈ch穗on蘆dr杜ia村l責pe厚rm信ea拳bi怒li梳t(yī)y盲t魔ra揪ns召it說io剖n.A販sp剪ec侵ul卡at忘iv誼e資mo志de法l傾sh體ow設(shè)in遞g司so慚me第o擋f抖th信e箱co疫mp州on勇en思ts辨o得f繩th碼e扒pe癢rm駱ea慕bi且li殼ty彈t循ra園ns壁it當io肺n叢po浸re泡.柴Th迫e貨ro肝le多s殃ofpo恒ri色nan需d牲th討e抽be錄nz商od稈ia覆ze粗pi屯ne從r擱ec究ep釘to捎r重re癥ma滅in幟c升ir建cu去ms奔ta秧nt堪ia畫l.呆I其n閱th炒e踏op泊en壯c澤on層fi劑gu思ra伐ti側(cè)on維,稈wa誦te啟r桿an暑d氏so郵lu需te炒s鍬en絡te勾r積th販e賠ma填tr因ix秒,險ca田us重in片g斬ma或tr平ix脊s緊we嗓ll震in爭g已an雖d襲ou眠te諒r睛me摟mb皮ra牌ne做d叢is擁ru鍛pt償io寒n謹(s掌ee剩F鉗ig擠.吩1)霞,賊le窮ad岔i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