AbMole丨Mito-TEMPO：從分子機制到科研應(yīng)用的靶向線粒體抗氧化劑線粒體是真核細胞內(nèi)負責(zé)能量代謝和活性氧（ReactiveOxygenSpecies,ROS）生成的核心細胞器。生理狀態(tài)下，線粒體ROS參與細胞信號傳導(dǎo)；但當(dāng)出于氧化應(yīng)激時，線粒體DNA（mtDNA）、脂質(zhì)及蛋白質(zhì)會發(fā)生氧化損傷，進而導(dǎo)致線粒體功能紊亂，甚至細胞死亡（細胞鐵死亡和焦亡等）。傳統(tǒng)抗氧化劑難以精準作用于線粒體，Mito-TEMPO可彌補上述缺陷。Mito-TEMPO（AbMole，M10919）作為一種線粒體靶向型抗氧化劑，通過特定分子設(shè)計實現(xiàn)了自身在線粒體中的高效富集，為研究線粒體ROS的功能及調(diào)控機制提供了關(guān)鍵工具。一、Mito-TEMPO的分子結(jié)構(gòu)與線粒體靶向機制Mito-TEMPO（AbMole，M10919）的核心結(jié)構(gòu)是TEMPO（2,2,6,6-四甲基哌啶-1-氧自由基）和TPP+（三苯基膦）。TEMPO（AbMole，M11341）是一種ROS清除劑，具有超氧化物歧化酶（SOD）模擬活性，可以分解超氧陰離子在內(nèi)的多種ROS。Mito-TEMPO是在TEMPO基礎(chǔ)上共價連接TPP+形成的。TPP+具有線粒體靶向結(jié)構(gòu)，首先TPP+的親脂性使其能夠穿透生物膜；其次TPP+的正電荷可通過線粒體膜電位（ΔΨm）滯留在線粒體基質(zhì)中。有文獻報導(dǎo)線粒體膜電位（-150至-180mV）為帶正電的TPP+提供了電化學(xué)梯度，使Mito-TEMPO在線粒體基質(zhì)中的濃度可比胞質(zhì)中高100-500倍ADDINEN.CITEADDINEN.CITE.DATA[1]。圖SEQ圖\*ARABIC1.線粒體中ROS的產(chǎn)生機制ADDINEN.CITEADDINEN.CITE.DATA[2]二、Mito-TEMPO的研究應(yīng)用1.Mito-TEMP用于清除線粒體ROSMito-TEMPO（AbMole，M10919）主要通過清除線粒體活性氧(mtROS)發(fā)揮其生物學(xué)效應(yīng)。在多種實驗體系中，Mito-TEMPO處理可顯著降低mtROS水平，改善線粒體膜電位(MMP)去極化狀態(tài)，并恢復(fù)線粒體氧化磷酸化(OXPHOS)功能ADDINEN.CITEADDINEN.CITE.DATA[3]。例如Mito-TEMPO在糖尿病小鼠模型中，逆轉(zhuǎn)了高糖誘導(dǎo)的線粒體膜電位耗散和ROS積累ADDINEN.CITEADDINEN.CITE.DATA[4]。Mito-TEMPO在雙酚A（BPA）暴露的大鼠模型中，通過清除ROS保護睪丸組織中線粒體功能、減少線粒體氧化應(yīng)激、維持線粒體膜電位（MMP）和線粒體復(fù)合物II和IV的酶活性ADDINEN.CITE<EndNote><Cite><Author>Shetty</Author><Year>2022</Year><RecNum>907</RecNum><DisplayText><styleface="superscript">[5]</style></DisplayText><record><rec-number>907</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756196686">907</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Shetty,S.</author><author>Kumar,V.</author><author>Ramesh,V.</author><author>Bharati,S.</author></authors></contributors><auth-address>DepartmentofNuclearMedicine,ManipalCollegeofHealthProfessions,ManipalAcademyofHigherEducation,Manipal,India.</auth-address><titles><title>Mito-TEMPOprotectsagainstbisphenol-A-inducedtesticulartoxicity:anin

vivostudy</title><secondary-title>FreeRadicRes</secondary-title><alt-title>Freeradicalresearch</alt-title></titles><periodical><full-title>FreeRadicRes</full-title><abbr-1>Freeradicalresearch</abbr-1></periodical><alt-periodical><full-title>FreeRadicRes</full-title><abbr-1>Freeradicalresearch</abbr-1></alt-periodical><pages>427-435</pages><volume>56</volume><number>5-6</number><edition>2022/10/08</edition><keywords><keyword>Humans</keyword><keyword>Male</keyword><keyword>Rats</keyword><keyword>Animals</keyword><keyword>*Testis</keyword><keyword>ReactiveOxygenSpecies/pharmacology</keyword><keyword>Rats,Wistar</keyword><keyword>*Semen</keyword><keyword>Antioxidants/pharmacology</keyword><keyword>OxidativeStress</keyword><keyword>Testiculardamage</keyword><keyword>bisphenol-A(BPA)</keyword><keyword>mitochondriadysfunction</keyword><keyword>mitochondria-targetedantioxidant</keyword><keyword>mitochondrialoxidativestress</keyword></keywords><dates><year>2022</year><pub-dates><date>May-Jun</date></pub-dates></dates><isbn>1029-2470</isbn><accession-num>36205519</accession-num><urls></urls><electronic-resource-num>10.1080/10715762.2022.2133702</electronic-resource-num><remote-database-provider>NLM</remote-database-provider><language>eng</language></record></Cite></EndNote>[5]。Mito-TEMPO還能降低5-FU（5-Fluorouracil）誘導(dǎo)的mtROS和mtLPO（線粒體脂質(zhì)過氧化物），以改善線粒體復(fù)合物II的活性ADDINEN.CITEADDINEN.CITE.DATA[6]。Mito-TEMPO用于ROS相關(guān)的信號通路調(diào)控Mito-TEMPO（AbMole，M10919）的作用機制不僅限于簡單的ROS清除，它還能調(diào)節(jié)多種線粒體相關(guān)信號通路和細胞生命活動。例如Mito-TEMPO在MCF-7細胞中，逆轉(zhuǎn)了DHA誘導(dǎo)的ROS依賴性自噬激活，通過調(diào)節(jié)p-AMPKα/p-Raptor和p-mTOR/p-ULK1通路影響自噬體形成ADDINEN.CITEADDINEN.CITE.DATA[7]。在使用Mito-TEMPO處理MIN6細胞后，發(fā)現(xiàn)Mito-TEMPO可激活PINK1和Parkin通路促進線粒體自噬，并減少鐵死亡ADDINEN.CITEADDINEN.CITE.DATA[8]。Mito-TEMPO還可通過增強SOD活性和PI3K/AKT/mTOR磷酸化發(fā)揮神經(jīng)保護作用ADDINEN.CITE<EndNote><Cite><Author>Mukem</Author><Year>2021</Year><RecNum>913</RecNum><DisplayText><styleface="superscript">[9]</style></DisplayText><record><rec-number>913</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756197834">913</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Mukem,S.</author><author>Thongbuakaew,T.</author><author>Khornchatri,K.</author></authors></contributors><auth-address>SchoolofMedicine,WalailakUniversity,NakhonSiThammarat,80160,Thailand. ChulabhornInternationalCollegeofMedicine,ThammasatUniversity,RangsitCampus,Pathumthani,12120,Thailand.</auth-address><titles><title>Mito-TemposuppressesautophagicfluxviathePI3K/Akt/mTORsignalingpathwayinneuroblastomaSH-SY5Ycells</title><secondary-title>Heliyon</secondary-title><alt-title>Heliyon</alt-title></titles><periodical><full-title>Heliyon</full-title><abbr-1>Heliyon</abbr-1></periodical><alt-periodical><full-title>Heliyon</full-title><abbr-1>Heliyon</abbr-1></alt-periodical><pages>e07310</pages><volume>7</volume><number>6</number><edition>2021/07/02</edition><keywords><keyword>Autophagy</keyword><keyword>Excitotoxicity</keyword><keyword>Glutamate</keyword><keyword>Mito-Tempo</keyword><keyword>Mitochondrialdysfunction</keyword><keyword>Oxidativestress</keyword></keywords><dates><year>2021</year><pub-dates><date>Jun</date></pub-dates></dates><isbn>2405-8440(Print) 2405-8440</isbn><accession-num>34195421</accession-num><urls></urls><custom2>PMC8239474</custom2><electronic-resource-num>10.1016/j.heliyon.2021.e07310</electronic-resource-num><remote-database-provider>NLM</remote-database-provider><language>eng</language></record></Cite></EndNote>[9]。Mito-TEMPO用于程序性細胞死亡的研究鐵死亡(Ferroptosis)：多項研究表明Mito-TEMPO（AbMole，M10919）能夠有效抑制鐵死亡過程。Mito-TEMPO在三氯乙烯(TCE)誘導(dǎo)小鼠腎小管上皮細胞的鐵死亡模型中，阻斷了鐵蛋白降解和鐵死亡的發(fā)生ADDINEN.CITEADDINEN.CITE.DATA[10]。類似地，Mito-TEMPO在Andrographolide（ADE）誘導(dǎo)的非小細胞肺癌細胞鐵死亡模型中，通過改善線粒體功能障礙，抑制了ADE誘導(dǎo)的鐵死亡。這些發(fā)現(xiàn)提示線粒體ROS是鐵死亡發(fā)生的關(guān)鍵調(diào)控因素ADDINEN.CITEADDINEN.CITE.DATA[11]。細胞焦亡(Pyroptosis)：Mito-TEMPO（AbMole，M10919）也常用于細胞凋亡的研究。例如Mito-TEMPO在苯并芘(BaP)誘導(dǎo)的肝細胞損傷研究中，起到與NLRP3抑制劑MCC950類似的作用，能夠減弱BaP對Caspase-1依賴性焦亡途徑的激活。此外，Mito-TEMPO還可顯著抑制動脈粥樣硬化動物模型中ox-LDL觸發(fā)的NLRP3炎癥小體激活A(yù)DDINEN.CITEADDINEN.CITE.DATA[12]。Mito-TEMPO用于維持線粒體結(jié)構(gòu)與功能完整性Mito-TEMPO保護氧化磷酸化（OXPHOS）系統(tǒng)：ROS過量會直接損傷線粒體電子傳遞鏈（ETC）的復(fù)合物，抑制氧化磷酸化。Mito-TEMPO（AbMole，M10919）通過靶向清除線粒體ROS，減輕了這種抑制。例如，在脂多糖（LPS）誘導(dǎo)的小鼠膿毒癥模型中，Mito-TEMPO預(yù)處理通過降低MDA含量（氧化應(yīng)激標志物）和提升SOD活性，恢復(fù)了線粒體大小和功能，支持了氧化磷酸化的正常進行ADDINEN.CITE<EndNote><Cite><Author>Wang</Author><Year>2022</Year><RecNum>919</RecNum><DisplayText><styleface="superscript">[13]</style></DisplayText><record><rec-number>919</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756200294">919</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Wang,P.F.</author><author>Xie,K.</author><author>Cao,Y.X.</author><author>Zhang,A.</author></authors></contributors><auth-address>DepartmentofCriticalCareMedicine,TheSecondAffiliatedHospitalofChongqingMedicalUniversity,Chongqing,China.</auth-address><titles><title>HepatoprotectiveEffectofMitochondria-TargetedAntioxidantMito-TEMPOagainstLipopolysaccharide-InducedLiverInjuryinMouse</title><secondary-title>MediatorsInflamm</secondary-title><alt-title>Mediatorsofinflammation</alt-title></titles><periodical><full-title>MediatorsInflamm</full-title><abbr-1>Mediatorsofinflammation</abbr-1></periodical><alt-periodical><full-title>MediatorsInflamm</full-title><abbr-1>Mediatorsofinflammation</abbr-1></alt-periodical><pages>6394199</pages><volume>2022</volume><edition>2022/07/01</edition><keywords><keyword>Animals</keyword><keyword>Antioxidants/metabolism</keyword><keyword>*ChemicalandDrugInducedLiverInjury,Chronic</keyword><keyword>CyclicN-Oxides</keyword><keyword>Lipopolysaccharides/pharmacology</keyword><keyword>Mice</keyword><keyword>Mitochondria/metabolism</keyword><keyword>OxidativeStress</keyword><keyword>ReactiveOxygenSpecies/metabolism</keyword><keyword>*Sepsis/metabolism</keyword></keywords><dates><year>2022</year></dates><isbn>0962-9351(Print) 0962-9351</isbn><accession-num>35769207</accession-num><urls></urls><custom2>PMC9236847</custom2><electronic-resource-num>10.1155/2022/6394199</electronic-resource-num><remote-database-provider>NLM</remote-database-provider><language>eng</language></record></Cite></EndNote>[13]。Mito-TEMPO維持線粒體膜電位（MMP）：線粒體膜電位是驅(qū)動氧化磷酸化的關(guān)鍵因素。Mito-TEMPO（AbMole，M10919）在多種模型中（如神經(jīng)細胞、精子冷凍保存、肝損傷等研究均表現(xiàn)出提升MMP的作用。例如，Mito-TEMPO在紫外模擬的Wistar大鼠皮膚損傷模型中，不僅減少了ROS生成，還顯著提高了線粒體膜電位（MMP）ADDINEN.CITEADDINEN.CITE.DATA[14]。范例詳解PesticideBiochemistryandPhysiology.2024Apr27.東北農(nóng)業(yè)大學(xué)的研究團隊在該論文中探究了甲草胺（MET）對草魚肝細胞（L8824細胞）的毒性機制，以及褪黑素（Melatonin，MT）對甲草胺的拮抗作用。通過體外實驗與計算機模擬分析發(fā)現(xiàn)如下機制：MET暴露會誘導(dǎo)L8824細胞死亡，且焦亡是主要死亡形式。具體表現(xiàn)為細胞腫脹、細胞膜破裂、胞內(nèi)容物釋放，同時伴隨乳酸脫氫酶（LDH）釋放增加、細胞活力下降。MET主要通過ROS破壞線粒體功能（線粒體膜電位下降，線粒體DNA釋放），并激活NLRP3-ASC炎癥小體/Caspase-1通路，進而切割GSDMD，引發(fā)焦亡炎癥反應(yīng)，形成“ROS-NLRP3-Caspase-1-焦亡”的循環(huán)放大效應(yīng)。來自AbMole的Mito-TEMPO（AbMole，M10919）作為線粒體特異性抗氧化劑，在實驗中主要用于驗證“線粒體ROS是MET誘導(dǎo)焦亡的關(guān)鍵介質(zhì)”這一機制。實驗表明，Mito-TEMPO預(yù)處理可顯著減少ROS生成，恢復(fù)線粒體膜電位，降低8-OHdG水平，抑制NLRP3-ASC炎癥小體激活及下游炎癥因子（如IL-1β、IL-18、TNF-α和IL-6）的表達，從而減輕MET誘導(dǎo)的焦亡炎癥反應(yīng)。圖SEQ圖\*ARABIC2.METexposureactivatesNLRP3-ASCinflammasomepathwayofL8824cells.參考文獻ADDINEN.REFLIST[1]S.Shetty,U.Anushree,R.Kumar,etal.,Mitochondria-targetedantioxidant,mito-TEMPOmitigatesinitiationphaseofN-Nitrosodiethylamine-inducedhepatocarcinogenesis,Mitochondrion58(2021)123-130.[2]J.P.Mazat,A.Devin,S.Ransac,ModellingmitochondrialROSproductionbytherespiratorychain,Cellularandmolecularlifesciences:CMLS77(3)(2020)455-465.[3]H.Wu,T.Xu,N.Yang,etal.,Low-SeDietIncreasedMitochondrialROStoSuppressMyoblastsProliferationandPromoteApoptosisinBroilersviamiR-365-3p/SelTSignalingAxis,Journalofagriculturalandfoodchemistry72(1)(2024)284-299.[4]L.Huang,Z.Chen,R.Chen,etal.,Increasedfattyacidmetabolismattenuatescardiacresistancetoβ-adrenoceptoractivationviamitochondrialreactiveoxygenspecies:Apotentialmechanismofhypoglycemia-inducedmyocardialinjuryindiabetes,Redoxbiology52(2022)102320.[5]S.Shetty,V.Kumar,V.Ramesh,etal.,Mito-TEMPOprotectsagainstbisphenol-A-inducedtesticulartoxicity:anin

vivostudy,Freeradicalresearch56(5-6)(2022)427-435.[6]P.K.Tambe,H.S.Qsee,S.Bharati,Mito-TEMPOmitigates5-fluorouracil-inducedintestinalinjuryviaattenuatingmitochondrialoxidativestress,inflammation,andapoptosis:aninvivostudy,Inflammopharmacology31(4)(2023)2091-2102.[7]C.H.Tsai,C.K.Lii,T.S.Wang,etal.,DocosahexaenoicacidpromotestheformationofautophagosomesinMCF-7breastcancercellsthroughoxidativestress-inducedgrowthinhibitor1mediatedactivationofAMPK/mTORpathway,Foodandchemicaltoxicology:aninternationaljournalpublishedfortheBritishIndustrialBiologicalResearchAssociation154(2021)112318.[8]B.Chang,Y.Su,T.Li,etal.,Mito-TEMPOAmelioratesSodiumPalmitateInducedFerroptosisinMIN6CellsthroughPINK1/Parkin-MediatedMitophagy,Biomedicalandenvironmentalsciences:BES37(10)(2024)1128-1141.[9]S.Mukem,T.Thongbuakaew,K.Khornchatri,Mito-TemposuppressesautophagicfluxviathePI3K/Akt/mTORsignalingpat