背景與目的:腎缺血再灌注損傷（Renal Ischemia/Reperfusion Injury）是引起急性腎損傷（acute kidney injury,AKI）的主要原因之一。間充質(zhì)干細(xì)胞（mesenchymal stem cells,MSCs）的細(xì)胞外囊泡（Extracellular Vesicles,EVs）應(yīng)用于多種疾病模型的干預(yù)治療上,其主要作用中包括減輕氧化應(yīng)激和恢復(fù)線粒體功能,但其中關(guān)鍵有效成分尚不明確。我們嘗試應(yīng)用人臍帶間充質(zhì)干細(xì)胞的細(xì)胞外囊泡對(duì)小鼠腎缺血再灌注損傷進(jìn)行治療的驗(yàn)證;同時(shí)應(yīng)用蛋白組學(xué)和生物信息學(xué)分析,篩選出細(xì)胞外囊泡中可能起治療作用的關(guān)鍵組份,希望獲得的結(jié)果為未來深入研究打下基礎(chǔ)。方法:從人臍帶分離華通膠部分原代培養(yǎng)間充質(zhì)干細(xì)胞,培養(yǎng)至細(xì)胞生長(zhǎng)80%-90%融合時(shí)更換無血清培養(yǎng)基,24小時(shí)后收取條件培養(yǎng)基利用超速離心法提取細(xì)胞外囊泡。對(duì)提取獲得的細(xì)胞外囊泡進(jìn)行透射電子顯微鏡（Transmission Electron Microscope,TEM）和納米流式檢測(cè)儀（NanoFCM）鑒定。建立左腎缺血25min,右腎切除的小鼠模型。隨機(jī)分為未IR損傷組,I... 

【文章頁數(shù)】：74 頁

【學(xué)位級(jí)別】：博士

【文章目錄】：
摘要
ABSTRACT
中文部分
    緒論
    材料與方法
        1.實(shí)驗(yàn)試劑和儀器
        2.人臍帶間充質(zhì)干細(xì)胞和成纖維細(xì)胞的體外分離及培養(yǎng)
        3.提取及鑒定人臍帶間充質(zhì)干細(xì)胞的細(xì)胞外囊泡
        4.動(dòng)物實(shí)驗(yàn)
        5.數(shù)據(jù)分析與統(tǒng)計(jì)學(xué)處理
    結(jié)果
        1.人臍帶間充質(zhì)干細(xì)胞的體外培養(yǎng)及細(xì)胞外囊泡的鑒定
        2.人臍帶間充質(zhì)干細(xì)胞的細(xì)胞外囊泡的濃度測(cè)定及粒徑檢測(cè)
        3.小鼠腎缺血再灌注損傷造模及干預(yù)后腎功能變化情況
        4.小鼠腎缺血再灌注損傷造模及干預(yù)后腎形態(tài)學(xué)的評(píng)估
        5.小鼠腎缺血再灌注損傷造模及干預(yù)后腎組織細(xì)胞凋亡情況檢測(cè)
        6.間充質(zhì)干細(xì)胞和成纖維細(xì)胞兩種細(xì)胞來源的細(xì)胞外囊泡的蛋白組學(xué)分析
    討論
    結(jié)論
英文部分
    Introduction
    Materials and Methods
        1.Experimental reagents and instruments
        2.In vitro isolation and culture of human umbilical cord mesenchymal stem cells and fibroblasts
        3.Extraction and identification of extracellular vesicles of human umbilical cord mesenchymal stem cells
        4.Animal experiment
        5.Data analysis and statistical processing
    Results
        1.Culture of human umbilical cord mesenchymal stem cells in vitro and identification of extracellular vesicles
        2.Determination of the concentration of extracellular vesicles in human umbilical cord mesenchymal stem cells and particle size detection
        3.Modeling of renal ischemia-reperfusion injury in mice and changes of renal function after intervention
        4.Modeling of renal ischemia-reperfusion injury in mice and evaluation of renal morphology after intervention
        5.Detection of renal tissue apoptosis after modeling and intervention of renal ischemia-reperfusion injury in mice
        6.Proteomic analysis of extracellular vesicles derived from mesenchymal stem cells and fibroblasts
    Discussion
    Conclusion
參考文獻(xiàn)
致謝
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