AbMole詳解：PMA（佛波酯）在激活T細(xì)胞、誘導(dǎo)細(xì)胞分化、構(gòu)建炎癥模型上的應(yīng)用Phorbol12-myristate13-acetate（PMA，佛波酯，AbMole，M4647）是一種從植物中提取的二酯類(lèi)化合物，PMA的化學(xué)結(jié)構(gòu)使其能夠模擬細(xì)胞內(nèi)重要的信號(hào)分子二酰基甘油（DAG），進(jìn)而激活蛋白激酶C（PKC）。作用機(jī)制蛋白激酶C（ProteinKinaseC,PKC）是一類(lèi)依賴鈣和磷脂的絲氨酸/蘇氨酸蛋白激酶家族，PKC家族成員（如PKCα、PKCβ、PKCγ等經(jīng)典型PKC，以及PKCδ、PKCε等新型PKC）的結(jié)構(gòu)具有高度保守性（圖1），均包含調(diào)節(jié)結(jié)構(gòu)域和催化結(jié)構(gòu)域。調(diào)節(jié)結(jié)構(gòu)域含有DAG結(jié)合位點(diǎn)（C1結(jié)構(gòu)域）和鈣結(jié)合位點(diǎn)（C2結(jié)構(gòu)域），其催化結(jié)構(gòu)域負(fù)責(zé)底物磷酸化ADDINEN.CITEADDINEN.CITE.DATA[1]。二酰甘油（Diacylglycerol,DAG）是其重要的內(nèi)源性激活信號(hào)分子。DAG可作為第二信使，與PKC調(diào)節(jié)結(jié)構(gòu)域的C1結(jié)構(gòu)域結(jié)合，誘導(dǎo)PKC構(gòu)象改變，使其從胞質(zhì)轉(zhuǎn)位至細(xì)胞膜（或其他膜結(jié)構(gòu)），同時(shí)解除自身抑制性結(jié)構(gòu)域?qū)Υ呋钚缘囊种疲┞洞呋稽c(diǎn)，從而激活PKC。PMA（TPA，佛波酯，AbMole，M4647））可以模擬DAG的作用，從而激活PKC，實(shí)現(xiàn)對(duì)細(xì)胞多種生理功能的調(diào)節(jié)，如細(xì)胞分化、增殖和凋亡等。圖SEQ圖\*ARABIC1.PKC家族的分類(lèi)和結(jié)構(gòu)示意圖ADDINEN.CITEADDINEN.CITE.DATA[1]研究應(yīng)用PMA調(diào)節(jié)炎癥反應(yīng)和細(xì)胞因子合成，激活T細(xì)胞PMA（佛波酯，AbMole，M4647）可以通過(guò)激活上游的信號(hào)分子（如PKC），進(jìn)而激活NF-κB信號(hào)通路，并調(diào)節(jié)多個(gè)基因的表達(dá)，這些表達(dá)產(chǎn)物包括各類(lèi)細(xì)胞因子、趨化因子等，它們參與調(diào)控細(xì)胞的炎癥反應(yīng)和免疫應(yīng)答等過(guò)程。在單核細(xì)胞向巨噬細(xì)胞分化過(guò)程中，PMA可激活NF-κB信號(hào)通路，促進(jìn)與巨噬細(xì)胞功能相關(guān)的基因表達(dá)，如CD11b、CD14等，使細(xì)胞獲得巨噬細(xì)胞的表型和功能。PKC通路還可以通過(guò)影響AP-1、NFAT等通路以激活T淋巴細(xì)胞，使其進(jìn)入活化狀態(tài)（圖2）ADDINEN.CITE<EndNote><Cite><Author>Brezar</Author><Year>2015</Year><RecNum>301</RecNum><DisplayText><styleface="superscript">[2]</style></DisplayText><record><rec-number>301</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1749024529">301</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Brezar,V.</author><author>Tu,W.J.</author><author>Seddiki,N.</author></authors></contributors><auth-address>INSERMU955,équipe16andFacultédeMédecine,UniversitéParisEst,Créteil,France;VaccineResearchInstitute(VRI),Créteil,France. FacultyofEducation,Science,TechnologyandMaths,UniversityofCanberra,Canberra,ACT,Australia.</auth-address><titles><title>PKC-ThetainRegulatoryandEffectorT-cellFunctions</title><secondary-title>FrontImmunol</secondary-title><alt-title>Frontiersinimmunology</alt-title></titles><periodical><full-title>FrontImmunol</full-title><abbr-1>Frontiersinimmunology</abbr-1></periodical><alt-periodical><full-title>FrontImmunol</full-title><abbr-1>Frontiersinimmunology</abbr-1></alt-periodical><pages>530</pages><volume>6</volume><edition>2015/11/04</edition><keywords><keyword>Pkc-θ</keyword><keyword>effectorTcells</keyword><keyword>immuneinterventions</keyword><keyword>immunesynapse</keyword><keyword>regulatoryTcells</keyword></keywords><dates><year>2015</year></dates><isbn>1664-3224(Print) 1664-3224</isbn><accession-num>26528291</accession-num><urls></urls><custom2>PMC4602307</custom2><electronic-resource-num>10.3389/fimmu.2015.00530</electronic-resource-num><remote-database-provider>NLM</remote-database-provider><language>eng</language></record></Cite></EndNote>[2]，因此PMA也是一種高效的T細(xì)胞激活劑。圖SEQ圖\*ARABIC2.抗原呈遞細(xì)胞通過(guò)激活PKC活化T細(xì)胞ADDINEN.CITE<EndNote><Cite><Author>Brezar</Author><Year>2015</Year><RecNum>301</RecNum><DisplayText><styleface="superscript">[2]</style></DisplayText><record><rec-number>301</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1749024529">301</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Brezar,V.</author><author>Tu,W.J.</author><author>Seddiki,N.</author></authors></contributors><auth-address>INSERMU955,équipe16andFacultédeMédecine,UniversitéParisEst,Créteil,France;VaccineResearchInstitute(VRI),Créteil,France. FacultyofEducation,Science,TechnologyandMaths,UniversityofCanberra,Canberra,ACT,Australia.</auth-address><titles><title>PKC-ThetainRegulatoryandEffectorT-cellFunctions</title><secondary-title>FrontImmunol</secondary-title><alt-title>Frontiersinimmunology</alt-title></titles><periodical><full-title>FrontImmunol</full-title><abbr-1>Frontiersinimmunology</abbr-1></periodical><alt-periodical><full-title>FrontImmunol</full-title><abbr-1>Frontiersinimmunology</abbr-1></alt-periodical><pages>530</pages><volume>6</volume><edition>2015/11/04</edition><keywords><keyword>Pkc-θ</keyword><keyword>effectorTcells</keyword><keyword>immuneinterventions</keyword><keyword>immunesynapse</keyword><keyword>regulatoryTcells</keyword></keywords><dates><year>2015</year></dates><isbn>1664-3224(Print) 1664-3224</isbn><accession-num>26528291</accession-num><urls></urls><custom2>PMC4602307</custom2><electronic-resource-num>10.3389/fimmu.2015.00530</electronic-resource-num><remote-database-provider>NLM</remote-database-provider><language>eng</language></record></Cite></EndNote>[2]在一些免疫細(xì)胞如T細(xì)胞的體外研究中，PMA常與Ionomycin（離子霉素，AbMole，M3621）一起使用，誘導(dǎo)細(xì)胞合成細(xì)胞因子。如果進(jìn)一步結(jié)合莫能菌素（Monensin，AbMole，M6991）和BrefeldinA（布雷非德菌素A，AbMole，M2294）等，抑制細(xì)胞因子的胞外轉(zhuǎn)運(yùn)，或者限制新合成的蛋白向高爾基體的轉(zhuǎn)運(yùn)，則可將細(xì)胞因子保留在胞內(nèi)，最后運(yùn)用流式細(xì)胞術(shù)等方法即可檢測(cè)細(xì)胞因子的類(lèi)型，通過(guò)這種方法可評(píng)估免疫細(xì)胞的功能狀態(tài)，確定免疫細(xì)胞的分化類(lèi)型。PMA誘導(dǎo)細(xì)胞分化在細(xì)胞分化研究領(lǐng)域，PMA（佛波酯，AbMole，M4647）除了被廣泛用于誘導(dǎo)單核細(xì)胞向巨噬細(xì)胞分化以外，在一些神經(jīng)細(xì)胞系的研究中，PMA還可與GDNF（AbMole，M10344）、BDNF（AbMole，M11193）等細(xì)胞因子聯(lián)合使用，誘導(dǎo)神經(jīng)細(xì)胞的分化，使其表現(xiàn)出神經(jīng)元的形態(tài)和功能特征，如長(zhǎng)出軸突和樹(shù)突，表達(dá)神經(jīng)元特異性的標(biāo)志物等ADDINEN.CITE<EndNote><Cite><Author>CastilloBautista</Author><Year>2022</Year><RecNum>297</RecNum><DisplayText><styleface="superscript">[3]</style></DisplayText><record><rec-number>297</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1749019475">297</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>CastilloBautista,C.M.</author><author>Sterneckert,J.</author></authors></contributors><auth-address>CenterforRegenerativeTherapiesTUDresden(CRTD),TechnischeUniversit?t(TU)Dresden,Dresden,Germany. MedicalFacultyCarlGustavCarusofTUDresden,Dresden,Germany.</auth-address><titles><title>ProgressandchallengesindirectingthedifferentiationofhumaniPSCsintospinalmotorneurons</title><secondary-title>FrontCellDevBiol</secondary-title><alt-title>Frontiersincellanddevelopmentalbiology</alt-title></titles><periodical><full-title>FrontCellDevBiol</full-title><abbr-1>Frontiersincellanddevelopmentalbiology</abbr-1></periodical><alt-periodical><full-title>FrontCellDevBiol</full-title><abbr-1>Frontiersincellanddevelopmentalbiology</abbr-1></alt-periodical><pages>1089970</pages><volume>10</volume><edition>2023/01/24</edition><keywords><keyword>assembloids</keyword><keyword>directeddifferentiationofpluripotentstemcells</keyword><keyword>iPScells</keyword><keyword>inducedpluripotentstemcells</keyword><keyword>motorneurons</keyword><keyword>neurodegenerativediseasaes</keyword><keyword>organoids</keyword><keyword>commercialorfinancialrelationshipsthatcouldbeconstruedasapotential</keyword><keyword>conflictofinterest.</keyword></keywords><dates><year>2022</year></dates><isbn>2296-634X(Print) 2296-634x</isbn><accession-num>36684437</accession-num><urls></urls><custom2>PMC9849822</custom2><electronic-resource-num>10.3389/fcell.2022.1089970</electronic-resource-num><remote-database-provider>NLM</remote-database-provider><language>eng</language></record></Cite></EndNote>[3]。PMA用于構(gòu)建炎癥相關(guān)動(dòng)物模型在動(dòng)物研究中，PMA（佛波酯，AbMole，M4647）常被用于構(gòu)建炎癥相關(guān)模型。在小鼠模型中，局部涂抹或注射PMA能夠誘導(dǎo)皮膚炎癥反應(yīng)。PMA刺激可導(dǎo)致小鼠皮膚出現(xiàn)紅斑、水腫等炎癥癥狀，組織學(xué)檢查可見(jiàn)炎癥細(xì)胞浸潤(rùn)、表皮增厚等病理改變。這一模型模擬了皮膚炎癥疾病的部分病理過(guò)程，如銀屑病等。在其他組織器官的炎癥模型構(gòu)建中，PMA也有應(yīng)用，例如急性胰腺炎（AP）、慢性胰腺炎（CP）、神經(jīng)炎癥等模型。范例詳解Engineering41(2024)189-203.科研人員在上述文章中探究了丙酮酸激酶M2（PKM2）在非酒精性脂肪性肝炎（NASH）相關(guān)纖維化中的作用。生物信息學(xué)篩選和分析表明，PKM2在纖維化NASH的細(xì)胞中表達(dá)上調(diào)，尤其是在巨噬細(xì)胞中。巨噬細(xì)胞特異性PKM2敲除在三種不同NASH模型中均顯著改善肝臟炎癥和纖維化程度。從機(jī)制上講，PKM2依賴性糖酵解促進(jìn)了促炎巨噬細(xì)胞中NLRP3的激活，從而誘導(dǎo)肝星狀細(xì)胞激活和纖維化?？蒲腥藛T在實(shí)驗(yàn)中通過(guò)使用AbMole的PMA（佛波酯，AbMole，M4647）（20ng/mL）誘導(dǎo)THP-1（髓系白血病單核細(xì)胞）分化為巨噬細(xì)胞，以展開(kāi)上述研究ADDINEN.CITE<EndNote><Cite><Author>Qu</Author><Year>2024</Year><RecNum>299</RecNum><DisplayText><styleface="superscript">[4]</style></DisplayText><record><rec-number>299</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1749021703">299</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Qu,Hengdong</author><author>Zhang,Di</author><author>Liu,Junli</author><author>Deng,Jieping</author><author>Xie,Ruoyan</author><author>Zhang,Keke</author><author>Li,Hongmei</author><author>Tao,Ping</author><author>Wang,Genshu</author><author>Sun,Jian</author><author>Luo,OscarJunhong</author><author>Qu,Chen</author><author>Ye,Wencai</author><author>Hong,Jian</author></authors></contributors><titles><title>TherapeuticTargetingofPKM2AmelioratesNASHFibrosisProgressioninaMacrophage-SpecificandLiver-SpecificManner</title><secondary-title>Engineering</secondary-title></titles><periodical><full-title>Engineering</full-title></periodical><pages>189-203</pages><volume>41</volume><keywords><keyword>PyruvatekinaseM2</keyword><keyword>Macrophages</keyword><keyword>Nonparenchymalcells</keyword><keyword>Heteroduplexoligonucleotide</keyword><keyword>Nonalcoholicsteatohepatitis</keyword><keyword>Liverfibrosis</keyword></keywords><dates><year>2024</year><pub-dates><date>2024/10/01/</date></pub-dates></dates><isbn>2095-8099</isbn><urls><related-urls><url>/science/article/pii/S2095809924002650</url></related-urls></urls><electronic-resource-num>/10.1016/j.eng.2024.05.005</electronic-resource-num></record></Cite></EndNote>[4]。圖SEQ圖\*ARABIC3.PKM2-dependentglycolysispromotesPKRphosphorylationandNLRP3activationinmacrophagesADDINEN.CITE<EndNote><Cite><Author>Qu</Author><Year>2024</Year><RecNum>299</RecNum><DisplayText><styleface="superscript">[4]</style></DisplayText><record><rec-number>299</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1749021703">299</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Qu,Hengdong</author><author>Zhang,Di</author><author>Liu,Junli</author><author>Deng,Jieping</author><author>Xie,Ruoyan</author><author>Zhang,Keke</author><author>Li,Hongmei</author><author>Tao,Ping</author><author>Wang,Genshu</author><author>Sun,Jian</author><author>Luo,OscarJunhong</author><author>Qu,Chen</author><author>Ye,Wencai</author><author>Hong,Jian</author></authors></contributors><titles><title>TherapeuticTargetingofPKM2AmelioratesNASHFibrosisProgressioninaMacrophage-SpecificandLiver-SpecificManner</title><secondary-title>Engineering</secondary-title></titles><periodical><full-title>Engineering</full-title></periodical><pages>189-203</pages><volume>41</volume><keywords><keyword>PyruvatekinaseM2</keyword><keyword>Macrophages</keyword><keyword>Nonparenchymalcells</keyword><keyword>Heteroduplexoligonucleotide</keyword><keyword>Nonalcoholicsteatohepatitis</keyword><keyword>Liverfibrosis</keyword></keywords><dates><year>2024</year><pub-dates><date>2024/10/01/</date></pub-dates></dates><isbn>2095-8099</isbn><urls><related-urls><url>/science/article/pii/S2095809924002650</url></related-urls></urls><electronic-resource-num>/10.1016/j.eng.2024.05.005</electronic-resource-num></record></Cite></EndNote>[4]JImmunotherCancer.2023Oct;11(10):e007230.科研人員在上述文章中探究了膀胱癌（BLCA）中LRFN2（leucine-richrepeatandfibronectintype-IIIdomain-containingprotein2）在腫瘤微環(huán)境（TME）中的作用，以及它如何影響免疫系統(tǒng)對(duì)腫瘤的清除。研究發(fā)現(xiàn)，LRFN2在膀胱癌中通過(guò)抑制CD8+T細(xì)胞的浸潤(rùn)和功能轉(zhuǎn)變，導(dǎo)致BLCA對(duì)免疫檢查點(diǎn)抑制劑產(chǎn)生抗性。具體來(lái)說(shuō)，LRFN2通過(guò)減少促炎細(xì)胞因子和趨化因子的分泌，抑制CD8+T細(xì)胞的招募和功能轉(zhuǎn)變，從而形成非炎癥性腫瘤微環(huán)境，導(dǎo)致免疫耐受。在文中，AbMole的Phorbol12-myristat