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研究生： 謝佑翎
Hsieh, Yu-Ling
論文名稱： 乙醇與膽固醇對DPPC脂質體雙層膜特性之影響
Ethanol and Cholesterol Effects on Bilayer Characteristics of DPPC Liposomes
指導教授： 楊毓民
Yang, Yu-Min
學位類別： 碩士
Master
系所名稱： 工學院 - 化學工程學系
Department of Chemical Engineering
論文出版年： 2017
畢業學年度： 105
語文別： 中文
論文頁數： 101
中文關鍵詞： 脂質體 、雙層膜堅硬度 、螢光偏極化 、乙醇效應 、膽固醇效應
外文關鍵詞： liposomes, bilayer rigidity, fluorescence polarization technique, ethanol effect, cholesterol effect
相關次數： 點閱：129  下載：9 
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    • 本研究使用磷脂質DPPC作為主材料，混合不同濃度的乙醇與膽固醇，並以強制型製程製備穩定的脂質體，藉以探討乙醇、膽固醇個別添加與同時添加於脂質體時，對液胞粒徑、界面電位、雙層膜堅硬度及相轉移行為等物理特性的影響。
    探討乙醇、膽固醇個別添加的效應，實驗結果顯示，添加乙醇後，液胞粒徑無明顯變化，但界面電位會下降。添加膽固醇後，液胞粒徑會上升，界面電位則下降。以螢光偏極化偵測液胞雙層膜堅硬度及相轉移行為，可發現乙醇主要干擾雙層膜頭基區域的排列，使相轉移溫度下降；膽固醇主要影響雙層膜核心位置之碳鏈秩序，對雙層膜相轉移溫度影響不大，且在相轉移溫度之上與之下，膽固醇的添加會誘發有序液晶相的形成，對雙層膜的堅硬度呈現相反效應。
    探討乙醇、膽固醇同時添加的效應，實驗結果顯示，固定膽固醇濃度，乙醇的添加，對液胞粒徑影響無一致趨勢，但會使界面電位下降，且使雙層膜在相轉移溫度之上時流動性提升；當固定乙醇濃度，膽固醇的添加，會使液胞粒徑呈現先下降後持平的趨勢，界面電位則無顯著影響，且膽固醇對雙層膜堅硬度同樣出現相反效應。

    The aim of this study was to investigate the effect of ethanol and cholesterol, respectively and simultaneously, on bilayer characteristics of DPPC liposomes. Stable liposomes were fabricated by using lipid with various amounts of ethanol and cholesterol in water through a forced formation approach, and the physical properties of vesicles were investigated by size, zeta potential and fluorescence polarization. In this work, fluorescence polarization was measured at different temperatures and typical sigmoidal thermograms could be observed.
    Experimental results revealed that the sizes of vesicles didn't show consistent trend, but zeta potential would decrease with addition of ethanol. Fluorescence polarization showed that addition of ethanol is likely to disorder part of the lipid headgroup region and decrease of phase transition temperature. Addition of cholesterol would increase the sizes and decrease zeta potentials of vesicles. However, as the liposomes also contained high concentration of ethanol, the effect of cholesterol was quite different and showed no consistent trend. Fluorescence polarization showed that the addition of cholesterol had no significant effect on phase transition temperature of lipid bilayers, but it would lead to disordering and condensing effect on vesicular bilayer rigidity at different phases. The opposite effect of cholesterol attributed to the formation of liquid ordered phase, and it could also be observed from liposomes with high concentration of ethanol.

    摘要 I Extended Abstract II 致謝 IX 總目錄 X 表目錄 XIV 圖目錄 XVI 符號 XXII 第一章 緒論 1 1-1 前言 1 1-2 研究動機與目的 7 第二章 文獻回顧 9 2-1 脂質體的結構與型態 9 2-2 脂質體的形成 10 2-3 乙醇體 13 2-4 脂質體的物理穩定性 16 2-5 脂質體的化學穩定性 20 2-6 添加劑對液胞雙層膜的特性之影響 22 2-6-1 乙醇效應 22 2-6-2 膽固醇效應 25 2-7 螢光偏極化測量原理 32 第三章 實驗 38 3-1 實驗藥品 39 3-2 實驗儀器及裝置 40 3-2-1 超音波震盪分散裝置 40 3-2-2 動態雷射光散射法粒徑/界面電位分析儀 (Dynamic Light Scattering, DLS) 41 3-2-3 螢光分光光譜儀 (Fluorescence spectrometer) 45 3-2-4 電子控溫裝置 48 3-2-5 穿透式電子顯微鏡 49 3-3 實驗方法 50 3-3-1 DPPC脂質體的製備 50 3-3-2 粒徑分布、界面電位與液胞存活期的測量 51 3-3-3 液胞雙層膜之螢光偏極化的測定 53 3-3-4 穿透式電子顯微鏡的分析 55 第四章 結果與討論 56 4-1 使用螢光偏極化偵測液胞雙層堅硬度之實驗條件 57 4-1-1 如何將螢光分子正確導入液胞雙層膜 57 4-1-2 螢光分子濃度與液胞物理性質之關聯 63 4-1-3 DPH螢光分子在乙醇水溶液中之螢光強度 65 4-2 乙醇效應 68 4-2-1 添加乙醇對DPPC脂質體粒徑、界面電位與穩定性之影響 68 4-2-2 添加乙醇的DPPC脂質體其表面型態 71 4-2-3 添加乙醇對DPPC脂質體雙層膜堅硬度與相轉移行為之影響 72 4-3 膽固醇效應 73 4-3-1 添加膽固醇對DPPC脂質體粒徑、界面電位與穩定性之影響 73 4-3-2 添加膽固醇的DPPC脂質體其表面型態 76 4-3-3 添加膽固醇對DPPC脂質體雙層膜堅硬度與相轉移行為之影響 77 4-4 同時添加乙醇與膽固醇之效應 81 4-4-1 同時添加乙醇及膽固醇對DPPC脂質體粒徑、界面電位與穩定性之影響 81 4-4-2 同時添加乙醇及膽固醇的DPPC脂質體其表面型態 86 4-4-3 同時添加乙醇及膽固醇對DPPC脂質體雙層膜堅硬度與相轉移行為之影 87 第五章 結論與建議 88 5-1 結論 88 5-2 建議 90 參考文獻 91

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