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    Please use this identifier to cite or link to this item: http://ir.lib.ncu.edu.tw/handle/987654321/3886

    Title: 利用表面電漿共振儀及恆溫滴定微卡計探討微脂粒與類澱粉胜?交互作用之動力學及熱力學研究;Kinetics and Thermodynamics of Liposome & s-amyloid Interactions by Surface Plasmon Resonance and Isothermal Titration Microcalorimetry
    Authors: 邱秀玫;Hsiu-Mei Chiu
    Contributors: 化學工程與材料工程研究所
    Keywords: 表面電漿共振儀;類澱粉胜肽;阿茲海默症;微脂粒;;beta-amyloid;kinetics;surface plasmon resonance;enthalpy;liposome;Alzheimer's disease
    Date: 2005-06-02
    Issue Date: 2009-09-21 12:25:07 (UTC+8)
    Publisher: 國立中央大學圖書館
    Abstract: 類澱粉胜肽(ß-amyloid,Aß)被認為是引起阿茲海默症的主要原因。Aß是具有自我聚集能力的兩性胜肽,一但從單體聚集至纖維狀(fibril)的結構就會毒殺神經細胞,但到目前為止Aß是如何引發細胞毒性及疾病的作用機制仍然不清楚,所以本研究主要是探討Aß與微脂粒之交互作用,以不同培養時間的Aß與不同組成之微脂粒(DPPC、DPPG添加GM1及膽固醇)進行討論,以期能說明Aß與細胞膜之作用行為。整個研究的主題分為兩個部份,第一部份是利用圓二色光譜儀(CD)、螢光光譜儀(Fluorescence spectroscopy)及原子力顯微鏡(AFM)偵測Aß隨培養時間其結構的變化;第二部份是利用表面電漿共振儀(SPR)及恆溫滴定微卡計(ITC)探討Aß與微脂粒交互作用之動力學及熱力學。 在Aß結構的偵測方面,培養ㄧ天後於AFM的結果可看到纖維狀的形成,螢光及CD的結果也都可發現相同的現象。在Aß與微脂粒交互作用之動力學研究方面,培養零天的Aß與不同組成微脂粒作用其親和力大小為DPPG>DPPC/DPPG(75:25 molar ratio)>DPPC/GM1/膽固醇(5:3:2 molar ratio)>DPPG/20 mole%膽固醇>DPPC。由親和力強度可判斷培養零天的Aß與不同組成微脂粒交互作用主要是靠靜電作用力。並可發現Aß對於GM1有較高的親和力;膽固醇的添加會降低微脂粒的流動性而改變了Aß與微脂粒的交互作用,特別於吸附量與吸附後膜結構之維持。培養ㄧ天的Aß與微脂粒作用其親和力大小為DPPC>DPPG/20 mole%膽固醇>DPPC/DPPG(75:25 molar ratio)>DPPG>DPPC/GM1/cholesterol(5:3:2 molar ratio)。由親和力強度可判斷培養一天的Aß與不同組成微脂粒交互作用主要是靠疏水作用力。以上結果初步地說明了Aß與細胞膜之作用機制與Aß培養時間不同而有所改變。在Aß與微脂粒交互作用之熱力學研究方面,由熱量變化可以發現靜電作用力及疏水作用力會同時存在。 ß-amyloid(Aß)was believed to cause the prime factor of Alzheimer’s disease. However, the mechanism of the cytotoxicity and the disease caused by Aß is still unclear. The goal of this work is to study the interactions of various incubated time of Aß with designed liposomes. The objective of this investigation was achived by the following studies : The structural and aggrergation information of Aß by by circular dichroism spectroscopy(CD), by Thioflavin T fluorescence assay and monitored by atomic force microscopy(AFM). Futhermore, this investigation utilized surface plasmon resonance (SPR) and isothermal titration microcalorimetry (ITC) to measure the kinetics and binding enthalpy of Aß with the liposomes. Kinetics of interactions of Aß and liposomes by SPR reveal the driving force of fresh Aß interacts with various liposomes is electrostatics. And fresh Aß have high affinity with GM1. Addition of cholesterol to the liposome could alter membrane fluidity and affect the interactions of fresh Aß with liposomes especially in the amount of absorbed Aß and maintained the structure of liposome after adsorbing. The driving force of the 1 day of incubation of Aß interacts with various liposomes is hydrophobicity. The binding enthalpy measurements between Aß and liposomes by ITC are endothermic reaction. When the composition of liposome is zwitterionic lipids, the interaction of Aß with liposomes is predominantly hydrophobic force; in contrast with the drive force of interaction of charged lipid with Aß is electrostatic force.
    Appears in Collections:[化學工程與材料工程研究所] 博碩士論文

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