本文關(guān)鍵詞： 近紅外熒光成像 熒光小分子染料 細(xì)胞保護(hù) 干細(xì)胞示蹤 組織修復(fù)與再生　出處：《第三軍醫(yī)大學(xué)》2017年博士論文　論文類型：學(xué)位論文

【摘要】：研究背景干細(xì)胞移植治療在再生醫(yī)學(xué)領(lǐng)域受到越來(lái)越多的關(guān)注,但是由于對(duì)干細(xì)胞在移植后的遷移、歸巢、定植、分布等生物行為缺乏深刻的認(rèn)識(shí)和理解以及非常有限的體內(nèi)移植干細(xì)胞存活數(shù)量等原因,外源性干細(xì)胞移植治療效果還有待進(jìn)一步提高。因此,尋找一種能同時(shí)滿足當(dāng)前干細(xì)胞移植所面臨的細(xì)胞標(biāo)記示蹤和細(xì)胞保護(hù)兩方面需求的多功能材料,有利于促進(jìn)干細(xì)胞移植治療的發(fā)展。據(jù)檢索,目前還沒(méi)有這種多功能材料的相關(guān)報(bào)道。生物光學(xué)成像由于檢測(cè)儀器發(fā)展成熟、靈敏度高、對(duì)比度高、分辨率高、成像直觀、成像速度快和無(wú)損檢測(cè)等優(yōu)點(diǎn)被廣泛應(yīng)用于醫(yī)學(xué)生物研究。依據(jù)檢測(cè)方式的不同生物光學(xué)成像可分為熒光成像、生物發(fā)光成像、光聲成像、光學(xué)斷層層析成像等,其中活體動(dòng)物體內(nèi)光學(xué)成像主要采用生物發(fā)光成像與熒光成像。生物發(fā)光成像技術(shù)是用熒光素酶基因標(biāo)記細(xì)胞或DNA,而熒光成像技術(shù)則采用熒光報(bào)告基團(tuán)(GFP、RFP及Cyt等)或熒光染料進(jìn)行標(biāo)記。生物發(fā)光成像和部分熒光成像技術(shù)由于需要構(gòu)建基因表達(dá)載體轉(zhuǎn)染目標(biāo)細(xì)胞,具有潛在的生物風(fēng)險(xiǎn)性,導(dǎo)致其在臨床干細(xì)胞標(biāo)記成像中的應(yīng)用受到限制。而近紅外(near-infrared,NIR)熒光染料標(biāo)記技術(shù)由于較低的組織自發(fā)熒光和較強(qiáng)的組織穿透能力等特點(diǎn),通過(guò)熒光染料標(biāo)記可以對(duì)目標(biāo)細(xì)胞進(jìn)行實(shí)時(shí)、連續(xù)檢測(cè),在小動(dòng)物活體成像方面具有廣闊的應(yīng)用前景,目前正逐步應(yīng)用于細(xì)胞標(biāo)記示蹤研究。與量子點(diǎn)、碳點(diǎn)等無(wú)機(jī)近紅外熒光材料相比,有機(jī)近紅外熒光染料具有更高的熒光量子產(chǎn)率、結(jié)構(gòu)易調(diào)、生物安全性好等優(yōu)點(diǎn)而備受關(guān)注。但是,當(dāng)前已有的包括吲哚菁綠(indocyanine green,ICG)在內(nèi)的有機(jī)近紅外熒光染料大都是親脂性的細(xì)胞膜標(biāo)記染料,標(biāo)記效率較低,且在生物熒光成像過(guò)程中容易和體內(nèi)蛋白等生物分子相互作用,存在著嚴(yán)重的細(xì)胞沾染以及潛在的細(xì)胞毒性作用,限制了這類染料在干細(xì)胞標(biāo)記示蹤中的應(yīng)用。七甲川花菁染料是近紅外熒光染料中的優(yōu)秀代表,部分染料能夠特異性地蓄積于細(xì)胞線粒體之中并具有良好的光學(xué)穩(wěn)定性,多種七甲川花菁染料已經(jīng)應(yīng)用于生物活體中細(xì)胞標(biāo)記與示蹤成像研究。本課題組在之前的工作中,通過(guò)對(duì)七甲川花菁染料的化學(xué)結(jié)構(gòu)進(jìn)行修飾和改造,合成制備了一系列新型七甲川花菁熒光小分子化合物。通過(guò)結(jié)構(gòu)學(xué)研究分析以及其他研究者的相關(guān)報(bào)道,確定這類七甲川花菁熒光小分子能夠靶向蓄積于細(xì)胞線粒體之中;同時(shí)由于保留了七甲川花菁染料的近紅外熒光特性,為用于細(xì)胞標(biāo)記、活體示蹤以及生物成像提供了基礎(chǔ)。此外,此類七甲川花菁熒光小分子靶向蓄積于細(xì)胞線粒體之中,對(duì)線粒體功能產(chǎn)生一定影響,從而使細(xì)胞命運(yùn)發(fā)生轉(zhuǎn)變�；诋�(dāng)前干細(xì)胞移植治療的迫切需求和本課題組以往的研究工作基礎(chǔ),發(fā)展出一種新的能夠同時(shí)具有細(xì)胞標(biāo)記示蹤和細(xì)胞保護(hù)作用的七甲川花菁熒光小分子化合物,在干細(xì)胞治療領(lǐng)域具有重要意義和潛在應(yīng)用前景。研究方法1.多功能熒光小分子的合成與鑒定在以往的工作基礎(chǔ)上,通過(guò)對(duì)七甲川花菁染料的七甲川核心周邊N-烷基側(cè)鏈進(jìn)行結(jié)構(gòu)修飾和改造,合成制備了一系列新的、保留了七甲川花菁染料細(xì)胞線粒體靶向蓄積和近紅外熒光成像等原有特性的熒光小分子化合物。本研究從這些新型熒光小分子化合物中篩選出一種水溶性、熒光強(qiáng)度以及生物相容性較好,適用于干細(xì)胞示蹤成像,同時(shí)還能夠促進(jìn)細(xì)胞表達(dá)抗氧化相關(guān)蛋白,增強(qiáng)細(xì)胞抗氧化應(yīng)激損傷能力的多功能熒光小分子(NIR cell protector-61,NIRCP-61)。2.NIRCP-61用于間充質(zhì)干細(xì)胞標(biāo)記示蹤成像利用原代分離培養(yǎng)的四種不同來(lái)源的間充質(zhì)干細(xì)胞(mesenchymal stem/stromal cells,MSCs),包括人臍帶間充質(zhì)干細(xì)胞(h UCMSCs)、人真皮間充質(zhì)干細(xì)胞(h DMSCs)、大鼠真皮間充質(zhì)干細(xì)胞(r DMSCs)和小鼠骨髓間充質(zhì)干細(xì)胞(m BMMSCs),通過(guò)近紅外熒光顯微鏡觀察和流式細(xì)胞術(shù)(flow cytometry,FCM)檢測(cè)確定NIRCP-61對(duì)MSCs的最佳標(biāo)記條件和標(biāo)記效率;分別采用CCK-8實(shí)驗(yàn)、Annexin V/碘化丙啶(propidium iodide,PI)細(xì)胞凋亡檢測(cè)實(shí)驗(yàn)、細(xì)胞遷移實(shí)驗(yàn)、成骨誘導(dǎo)分化實(shí)驗(yàn)、成脂誘導(dǎo)分化實(shí)驗(yàn)檢測(cè)在最佳標(biāo)記條件下NIRCP-61對(duì)MSCs正常生理功能的影響;利用近紅外熒光顯微鏡檢測(cè)NIRCP-61標(biāo)記細(xì)胞后6天內(nèi)的熒光穩(wěn)定性并使用激光共聚焦檢測(cè)技術(shù)確定NIRCP-61標(biāo)記在細(xì)胞內(nèi)的亞細(xì)胞定位;建立放創(chuàng)復(fù)合傷小鼠模型,利用流式細(xì)胞術(shù)、激光共聚焦檢測(cè)技術(shù)和小動(dòng)物活體成像技術(shù),通過(guò)體外細(xì)胞實(shí)驗(yàn)和體內(nèi)動(dòng)物實(shí)驗(yàn)分別檢測(cè)NIRCP-61標(biāo)記MSCs對(duì)周邊非標(biāo)記細(xì)胞和組織的沾染情況,以及標(biāo)記細(xì)胞活體示蹤成像情況。3.NIRCP-61對(duì)間充質(zhì)干細(xì)胞抗氧化應(yīng)激損傷的實(shí)驗(yàn)研究通過(guò)過(guò)氧化氫(hydrogen peroxide,H2O2)處理或γ射線照射細(xì)胞,建立體外細(xì)胞氧化應(yīng)激損傷模型,采用CCK-8實(shí)驗(yàn)、Annexin V/PI細(xì)胞凋亡檢測(cè)實(shí)驗(yàn)和鈣黃綠素/PI染色檢測(cè)評(píng)價(jià)NIRCP-61減少H2O2處理造成的細(xì)胞活力降低、凋亡和死亡情況;采用集落形成實(shí)驗(yàn)、細(xì)胞核內(nèi)γ-H2AX免疫熒光染色、中性彗星實(shí)驗(yàn)以及Western Blot檢測(cè)評(píng)估NIRCP-61減輕電離輻射(ionizing radiation,IR)對(duì)細(xì)胞的損傷情況;采用流式細(xì)胞術(shù)和Western Blot檢測(cè)NIRCP-61處理后細(xì)胞和線粒體活性氧(reactive oxygen species,ROS)生成以及細(xì)胞內(nèi)抗氧化相關(guān)蛋白的表達(dá)變化情況;通過(guò)Western Blot檢測(cè)NIRCP-61處理后細(xì)胞內(nèi)核因子2相關(guān)因子2/抗氧化反應(yīng)元件(nuclear factorerythroid 2-related factor 2/antioxidant response element,Nrf2/ARE)和磷脂酰肌醇3-激酶/蛋白激酶B(phosphoinositide 3-kinase/proteinkinase B,PI3K/AKT)信號(hào)通路相關(guān)蛋白的表達(dá)變化情況;使用人Nrf2的小干擾RNA(small interfering RNA,si RNA)以及PI3K的抑制劑LY294002反向驗(yàn)證Nrf2/ARE和PI3K/AKT信號(hào)通路在NIRCP-61促進(jìn)細(xì)胞抗氧化應(yīng)激損傷中發(fā)揮的作用。4.NIRCP-61促進(jìn)間充質(zhì)干細(xì)胞體內(nèi)定植存活和組織修復(fù)的研究建立放創(chuàng)復(fù)合傷Sprague-Dawley(SD)大鼠模型和大劑量皮膚放射損傷大鼠模型,通過(guò)檢測(cè)創(chuàng)面愈合速度、瘢痕形成質(zhì)量、膠原形成情況以及觀察皮膚水皰、脫屑以及壞死性潰瘍形成等指標(biāo)評(píng)估NIRCP-61促進(jìn)間充質(zhì)干細(xì)胞定植存活,減輕機(jī)體組織損傷,促進(jìn)組織修復(fù)再生,提高細(xì)胞治療潛能的情況;通過(guò)人工結(jié)扎大鼠冠狀動(dòng)脈左前降支建立急性心肌梗死大鼠模型,分組治療4周后進(jìn)行超聲心動(dòng)圖檢測(cè)、心臟取材固定后進(jìn)行2,3,5-三苯基氯化四氮唑(2,3,5-Triphenyltetrazolium chloride,TTC)染色、免疫組織化學(xué)染色以及Masson染色檢測(cè)NIRCP-61促進(jìn)間充質(zhì)干細(xì)胞在大鼠體內(nèi)定植存活,減少左心室梗死面積,減輕組織纖維化形成,維持心臟正常功能的情況。研究結(jié)果1.通過(guò)對(duì)七甲川花菁染料進(jìn)行化學(xué)結(jié)構(gòu)修飾、改造,合成制備了一系列新型近紅外熒光小分子化合物,并從中成功篩選出新型熒光小分子(NIRCP-61),其水溶性和生物相容性好、標(biāo)記效率和熒光強(qiáng)度高,并且能夠同時(shí)滿足細(xì)胞移植治療對(duì)細(xì)胞標(biāo)記示蹤成像和細(xì)胞保護(hù)兩方面的需求。2.熒光小分子NIRCP-61保留了七甲川花菁染料良好的近紅外熒光以及細(xì)胞線粒體靶向蓄積的特性,體外細(xì)胞標(biāo)記過(guò)程簡(jiǎn)便易行,無(wú)需額外的轉(zhuǎn)染試劑或操作步驟即可完成。同當(dāng)前已有的其它近紅外熒光探針相比,NIRCP-61標(biāo)記效率高,生物相容性好,并且標(biāo)記細(xì)胞對(duì)周邊細(xì)胞和組織微環(huán)境的非特異性沾染明顯減少,顯著提升了熒光信號(hào)的特異性,在干細(xì)胞標(biāo)記示蹤成像中的應(yīng)用前景更佳。3.熒光小分子NIRCP-61能夠減輕過(guò)氧化氫處理造成的細(xì)胞活力降低,減少細(xì)胞凋亡;維持電離輻射后干細(xì)胞的成集落能力;免疫熒光染色、中性彗星實(shí)驗(yàn)以及Western Blot檢測(cè)結(jié)果證明NIRCP-61可明顯減少IR造成的細(xì)胞DNA損傷;ROS檢測(cè)結(jié)果顯示,NIRCP-61處理能顯著降低過(guò)氧化氫或IR處理引起的細(xì)胞內(nèi)ROS水平升高,減輕過(guò)量ROS對(duì)細(xì)胞造成的損傷;Western Blot檢測(cè)結(jié)果表明,NIRCP-61的細(xì)胞保護(hù)作用是通過(guò)活化細(xì)胞內(nèi)Nrf2和PI3K/Akt信號(hào)通路,激活細(xì)胞內(nèi)源性抗氧化還原反應(yīng)通路,促進(jìn)細(xì)胞高表達(dá)一系列內(nèi)源性抗氧化蛋白而實(shí)現(xiàn)。4.在三種動(dòng)物損傷模型中證明熒光小分子NIRCP-61能夠顯著減輕機(jī)體內(nèi)氧化應(yīng)激微環(huán)境對(duì)移植干細(xì)胞造成的損傷,促進(jìn)移植細(xì)胞在機(jī)體病損部位及周邊組織內(nèi)的定植和存活,減輕組織損傷和纖維化程度,加速創(chuàng)傷愈合,改善瘢痕形成質(zhì)量,促進(jìn)組織修復(fù)與再生,提高細(xì)胞移植治療的效果。研究結(jié)論據(jù)檢索,這種同時(shí)具備近紅外熒光成像和細(xì)胞保護(hù)功能的熒光小分子尚屬首次報(bào)道。線粒體靶向的NIRCP-61對(duì)不同種類的MSCs均具有優(yōu)異的標(biāo)記性能,適合用于干細(xì)胞的示蹤成像;且能顯著減輕氧化應(yīng)激對(duì)MSCs造成的損傷,促進(jìn)MSCs移植后在生物體內(nèi)的定植存活,減輕組織損傷,減少纖維化形成,促進(jìn)機(jī)體組織修復(fù)與再生,提高細(xì)胞治療潛能。因此,近紅外熒光小分子NIRCP-61代表了一類新穎的、可提高細(xì)胞移植治療效果的治療性熒光探針,在干細(xì)胞治療的臨床轉(zhuǎn)化中具有潛在的應(yīng)用前景。
[Abstract]:The research background of stem cell transplantation in the treatment has attracted more and more attention in the field of regenerative medicine, but due to the migration of stem cells in transplantation, homing, colonization, distribution of biological behavior and the lack of deep understanding and the understanding of stem cell transplantation in vivo very limited number of survival and other reasons, the treatment effect of exogenous stem cell transplantation remains to be further to improve. Therefore, looking for a can meet the current stem cell transplantation multifunctional material facing cell tracer and cell protection two aspects of demand, is conducive to the promotion of stem cell transplantation in the treatment of development. According to the search, there have been no reports of this kind of multifunctional materials. Because of bioluminescent imaging detection instrument development, high sensitivity, high contrast, high resolution imaging, intuitive, fast imaging and nondestructive testing is widely used in medical and biological research. According to the test Different biological optical imaging mode can be divided into fluorescence imaging, bioluminescence imaging, photoacoustic imaging, optical tomography tomography, the animal in vivo optical imaging mainly using bioluminescence imaging and fluorescence imaging. Bioluminescence imaging technology is the use of luciferase gene or DNA labeled cells, and fluorescence imaging using fluorescent (GFP, RFP and Cyt) or fluorescent dye labeled. Due to the construction of gene expression vector was transfected into target cell imaging and fluorescence imaging part of bioluminescence, with biological potential risk, resulting in the clinical application of stem cell marker imaging is limited. In the infrared (near-infrared, NIR) fluorescent dye labeled as technology the characteristics of low autofluorescence and strong tissue penetration, by fluorescent dye labeled target cells can be real-time, continuous inspection Test, has a broad application prospect in small animal imaging, is gradually being used in the study of cell labeling. Compared with quantum dots, carbon and other inorganic near infrared fluorescent materials, organic near infrared fluorescent dyes with higher fluorescence quantum yields, easy to adjust the structure, has the advantages of good bio security is attracting more and more attention however, the current including indocyanine green (indocyanine green, ICG), organic near infrared fluorescent dyes are cell membrane marker dye labeled lipophilic, low efficiency, and the interaction in biological fluorescence imaging process easily and in vivo protein and other biological molecules, there are serious contamination and cell cytotoxicity the potential and limits the type of dyes in the application of stem cell labeling in seven. Methine cyanine dye is an excellent representative of near infrared fluorescent dyes, dye specific volume In the mitochondria and has good optical stability, a variety of seven methine cyanine dyes have been applied to the study of cell biology in vivo labeling and tracing imaging. This group in the previous work, modified or altered by the chemical structure of seven methine cyanine dyes, a series of novel seven methine cyanine the fluorescence of small molecule compounds were synthesized. The related reports on analysis and other structural studies of the determination of such seven methine cyanine fluorescent molecules can accumulate in mitochondria targeting; at the same time due to the retention of the near infrared fluorescence of seven methine cyanine dyes, for cell labeling, provide based in vivo tracking and biological imaging. In addition, the seven methine cyanine fluorescent small molecules targeting accumulation in the mitochondria, have a certain impact on mitochondrial function, so that the cell fate occurred The current urgent needs and change. Do the research work based on previous group cell transplantation based on the development of a new can also have a cell labeling and cell protection seven methine cyanine fluorescent small molecule compounds, has important significance and potential application prospect in the field of stem cell therapy. Methods: 1. multifunctional fluorescent small molecule synthesis and identification based on the previous work, for structural modification through seven methine core surrounding the alkyl side chain of N- of seven methine cyanine dyes, the synthesis of a series of new compounds, fluorescent molecules retained seven a mitochondrial Sichuan cyanine dye cell targeting and accumulation of near infrared fluorescence imaging of the original characteristics. This study from these new fluorescent small molecules screened a water-soluble, fluorescence intensity and good biocompatibility, suitable for stem cells Cell imaging, but also can promote the expression of antioxidant related proteins in cells, enhance the multifunctional fluorescent small molecule cell oxidative stress damage ability (NIR cell protector-61, NIRCP-61.2.NIRCP-61) for mesenchymal stem cells labeled by imaging using four different sources of primary cultured mesenchymal stem cells (mesenchymal stem/stromal cells, MSCs), including human umbilical cord mesenchymal stem cells (H UCMSCs), human dermal mesenchymal stem cells (H DMSCs), rat dermal mesenchymal stem cells (R and DMSCs) of mouse bone marrow mesenchymal stem cells (m, BMMSCs) by near infrared fluorescence microscopy and flow cytometry. Surgery (flow cytometry, FCM NIRCP-61) testing to determine the optimal labeling conditions of MSCs and the labeling efficiency; using CCK-8 experiment, Annexin V/ propidium iodide (propidium iodide, PI) cell apoptosis assay, cell migration assay, bone induction The differentiation and adipogenic differentiation assay of NIRCP-61 in the best condition marked influence on MSCs of normal physiological function; the fluorescence stability of NIRCP-61 labeled cells 6 days after detection of near infrared fluorescence microscopy and confocal laser detection technique to determine the subcellular localization of NIRCP-61 labeled cells in mice; establishment of radiation combined wound the model, using flow cytometry, confocal laser detection technology and small animal imaging technology, NIRCP-61 marker MSCs on the surrounding non labeled cells and tissue contamination were detected by in vitro and in vivo animal experiments, and the experimental study of.3.NIRCP-61 labeled cells in vivo imaging of mesenchymal stem cells against oxidative stress injury by hydrogen peroxide (hydrogen peroxide, H2O2) or gamma ray irradiated cells, in vitro cell model establishment of oxidative stress injury, The CCK-8 experiment, the apoptosis of Annexin cells was determined by V/PI experiment and calcein /PI staining for detection and evaluation of NIRCP-61 H2O2 treatment caused a decrease in cell viability decreased, apoptosis and death; using colony formation assay, cell nucleus gamma -H2AX immunofluorescence staining, neutral comet assay and Western Blot detection and evaluation of NIRCP-61 reduced (ionizing radiation, IR of ionizing radiation) damage of cells; used after the treatment of NIRCP-61 cells and mitochondrial ROS was detected by flow cytometry and Western Blot (reactive oxygen species, ROS) generation and expression of related intracellular antioxidant; cell nuclear factor by Western Blot detection after treatment with NIRCP-61 2 related factors 2/ antioxidant response element (nuclear factorerythroid 2-related factor 2/antioxidant response element, Nrf2/ARE) and phosphatidylinositol 3- kinase / protein kinase B (Pho Sphoinositide 3-kinase/proteinkinase B, PI3K/AKT) expression of signal pathway related proteins; small interfering RNA use Nrf2 (small interfering RNA, Si RNA) and PI3K inhibitor LY294002 and PI3K/AKT reverse validation of Nrf2/ARE signal pathway in NIRCP-61 to promote the role of.4.NIRCP-61 in cell oxidative stress damage in promoting the survival of mesenchymal stem cells and tissues in vivo colonization of wound repair to establish the Sprague-Dawley (SD) rats model and large dose of radiation skin injury, wound healing through detection, the quality of scar formation, collagen formation and observation of skin blisters, desquamation and necrotic ulcer formation and evaluation index of NIRCP-61 promote mesenchymal stem cells transplanting survival, reduce the body tissue damage, promote tissue repair and regeneration, improve the therapeutic potential of the cells through the artificial ligation; In the left anterior descending coronary artery to establish acute myocardial infarction rat model group after 4 weeks of treatment, echocardiography, heart were fixed after 2,3,5- three 5-triphenyl tetrazolium chloride (2,3,5-Triphenyltetrazolium chloride, TTC) staining, immunohistochemical staining and Masson staining NIRCP-61 promote mesenchymal stem cell survival in vivo. In rats, reduce the infarct area of the left ventricle, reduce tissue fibrosis, maintain normal heart function. Results: 1. through chemical modification, modification of seven methine cyanine dyes, a series of novel near infrared fluorescent small molecule compounds were synthesized, and the successful screening of new fluorescent molecules (NIRCP-61), its water solubility and biocompatibility, fluorescence labeling efficiency and high strength, and can meet the need of cell transplantation for the treatment on cell labeling and imaging Cell protection requirements of.2. fluorescent molecules on NIRCP-61 two retained seven methine cyanine dyes with good NIR fluorescence and cell mitochondrial targeting accumulation characteristics, in vitro cell labeling process is simple and easy to complete, without additional transfection reagents or operation steps. Compared with other near infrared fluorescent probe with the existing. NIRCP-61 markers of high efficiency, good biocompatibility, and nonspecific contamination of labeled cells on the surrounding cells and tissue microenvironment significantly reduced, significantly enhance the specificity of fluorescence signal, the application prospect of stem.3. better fluorescent molecules of NIRCP-61 cells labeled by imaging can reduce the cell viability of hydrogen peroxide treatment caused a decrease in cell, reduce apoptosis; maintain after ionizing radiation stem cell colony forming ability; immunofluorescence staining, neutral comet assay and Western Blot assay results indicated that NIRCP -61 could significantly reduce DNA cell injury caused by IR; ROS assay showed that NIRCP-61 treatment can significantly reduce the elevated levels of ROS or IR treatment caused by hydrogen peroxide in cells, reduce excessive ROS causing damage to cells; Western Blot assay showed that NIRCP-61 cell protective effect by activating intracellular Nrf2 and PI3K/Akt signaling pathway activation of endogenous antioxidant, reaction pathway, and the realization of.4. in three kinds of animal models demonstrate the damage of fluorescent molecules of NIRCP-61 can significantly reduce the oxidative stress in the light microenvironment of the transplanted stem cells damage caused by the high expression of a series of endogenous antioxidant protein, promote colonization of transplanted cells in the lesion and surrounding tissues in the body and survival, reduce tissue damage and fibrosis, accelerate wound healing, improve the quality of scar formation, promote tissue repair and regeneration. High cell transplantation therapy. The research conclusion retrieval of fluorescent molecules with the near-infrared fluorescence imaging and cell protection function. This is the first report of NIRCP-61 mitochondrial targeting were tagged with excellent performance for different types of MSCs, suitable for stem cell imaging; and can significantly reduce the oxidative stress of damage caused by MSCs, survival in vivo to promote colonization of MSCs after transplantation, reduce tissue damage, reduce fibrosis, promote tissue repair and regeneration, improve the therapeutic potential of cells. Therefore, near infrared fluorescent small molecule NIRCP-61 represents a novel treatment, fluorescent probe can improve the efficacy of transplantation, with the potential application prospect in clinical translation of stem cell therapy.

【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R45

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