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研究生： 鄭嘉瑜
Cheng, Chia-Yu
論文名稱： 以螢光偏極化技術探討膽固醇對離子對雙親分子液胞雙層膜堅硬度的影響
A Study of Cholesterol Effect on Ion-Pair Amphiphile Vesicular Bilayer Rigidity by Using Fluorescence Polarization Technique
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： 2019
畢業學年度： 107
語文別： 中文
論文頁數： 111
中文關鍵詞： 離子對雙親分子 、陰陽離子液胞 、強制製程 、雙層膜堅硬度 、螢光偏極化技術 、膽固醇 、凝縮作用 、失序作用
外文關鍵詞： Ion-pair amphiphile, Catanionic vesicles, Mechanical dispersion fabrication process, Bilayer rigidity, Fluorescence polarization technique, Cholesterol, Condensing effect, Disordering effect.
相關次數： 點閱：137  下載：7 
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    • 　離子對雙親分子具有來源豐富、價格便宜、高化學穩定性及分子可設計性等優點，極有潛力做為製備類脂質體的材料。本研究運用螢光偏極化技術探討膽固醇添加劑對陰陽離子液胞雙層膜堅硬度的影響，及其對液胞穩定性與物理性質的效應。
    　　由螢光非等向性溫度圖譜的實驗結果顯示，膽固醇濃度的增加只會造成陰陽離子液胞雙層膜堅硬度的單調提升。此一效應與膽固醇對脂質體雙層膜堅硬度在高於及低於相轉移溫度時，呈現凝縮及失序的相反作用不同。推究原因可能是本研究所使用的離子對雙親分子 DeTMA-DS (decyltrimethylammonium-dodecylsulfate, C10-C12)、DeTMA-TS (decyltrimethylammonium-tetradecylsulfate, C10-C14)、DTMA-DS (dodecyltrimethylammonium-dodecylsulfate, C12-C12)，相較於脂質 dipalmitoylphosphatidylcholine (DPPC, C16-C16) 分子的碳鏈較短，具有較低的堅硬度，膽固醇對雙層膜的失序作用因而無從產生所致。此外，實驗結果也顯示，膽固醇的添加除了可以增進液胞穩定性之外，也使得經由強制製程形成的液胞粒徑隨著膽固醇濃度的增加而增大。本研究所獲得的這些膽固醇添加劑效應的影響結果對促進陰陽離子液胞做為藥物傳輸載體的可行性極有幫助。

    Due to their abundant sources, low cost, high chemical stability and molecular designability, lipid-like ion-pair amphipliles (IPAs, also known as catanionic surfactants) have emerged as attractive materials to prepare potential vesicular carriers in drug and gene delivery. In this work, cholesterol (CHOL) effect on bilayer rigidity of decyltrimethylammonium-dodecylsulfate, DeTMA-DS), decyltrimethylammonium-tetradecylsulfate (DeTMA-TS), and dodecyltrimethylammonium-dodecylsulfate (DTMA-DS) catanionic vesicles and dipalmitoylphosphatidylcholine (DPPC) liposomes were systematically studied by using fluorescence polarization technique. Vesicle stability and physical properties of them as the consequences of CHOL addition were also investigated.
    The experimental fluorescence anisotropy (FA) thermograms showed that fluorescence anisotropies of catanionic vesicle bilayers were monotonically increased with increasing of CHOL concentration. This effect of CHOL, however, is quite different from that on liposomal bilayers in which CHOL exhibited disordering and condensing effects when the temperatures were lower and higher, respectively, than the main phase transition temperature (Tm) of lipid. The different results are due to the fact that catanionic vesicles with thinner bilayer thickness exhibited much lower bilayer rigidity at temperatures lower than Tm as compared with those of liposomes. Under these conditions, there is no role for the disordering effect of CHOL to play on the ordering of bilayers. Furthermore, vesicle stability and size of vesicles fabricated by the process through the formation of thin films were found to be significantly enhanced with the addition of CHOL and reasonably explained by the increment of bilayer rigidity. The results achieved in this work is beneficial to applications of catanionic vesicles as carriers in drug and gene delivery.

    摘要 II Extended Abstract IV 致謝 XII 總目錄 XIII 表目錄 XVI 圖目錄 XVIII 符號 XXII 第一章 緒論 1 1-1 前言 1 1-2 研究動機與目的 9 第二章 文獻回顧 10 2-1 離子對雙親分子 10 2-2 陰陽離子液胞的形成 12 2-3 液胞的結構與型態 15 2-4 膽固醇效應 17 2-4-1 示差掃描熱卡分析(DSC) 19 2-4-2 螢光偏極化 (Fluorescence Polarization) 25 2-4-3膽固醇影響雙層膜相態之機制 27 2-5 螢光偏極化的測量原理 31 第三章 實驗 37 3-1 實驗藥品 38 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(Transmission Electron Microscope) 49 3-3 實驗方法 50 3-3-1 離子對雙親分子製備 50 3-3-2 IPA液胞製備 52 3-3-3 粒徑分布、界面電位與液胞存活期的測量 53 3-3-4 液胞雙層膜之螢光偏極化的測定 55 3-3-5 穿透式電子顯微鏡的分析 58 第四章 結果與討論 59 4-1 膽固醇對於IPA系統液胞物理性質及雙層膜堅硬度的影響 60 4-1-1 DeTMA-DS、DeTMA-TS及DTMA-DS液胞之物理性質 11111及雙層膜堅硬度 60 4-1-2 液胞曲率對於液胞雙層膜堅硬度的影響 74 4-1-3 液胞雙層膜堅硬度對於液胞形成粒徑影響 77 4-2 膽固醇效應對於IPA液胞與脂質DPPC液胞粒徑與雙層膜堅硬度的比較 80 4-2-1 膽固醇對於IPA系統及DPPC液胞粒徑的影響 80 4-2-2 膽固醇對於IPA系統及DPPC液胞雙層膜螢光非等向性的影響 82 4-2-3 IPA系統及DPPC系統雙層膜堅硬度差異 84 4-2-4 IPA系統及DPPC系統液胞粒徑與雙層膜堅硬度關聯性 92 第五章 結論與建議 94 5-1 結論 94 5-1-1膽固醇對於IPA液胞物理性質及雙層膜堅硬度影響 94 5-1-2膽固醇效應對於IPA液胞與脂質DPPC液胞粒徑與雙層膜堅硬度的比較 95 5-2 建議 96 參考文獻 97

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