本文選題：不對稱陽離子表面活性劑 + 陰-陽離子表面活性劑復(fù)配�。� 參考：《大連理工大學(xué)》2015年碩士論文

【摘要】：近年來,基因治療的研究一直是醫(yī)學(xué)界最重要的領(lǐng)域之一,其對于腫瘤、免疫缺陷癥等疾病已經(jīng)用于臨床；但是高毒性和相對較低的效率嚴(yán)重影響了其發(fā)展。相對于病毒載體的低安全性,非病毒載體的低免疫反應(yīng)、低毒和易制備等優(yōu)點使其成為研究熱點。本論文設(shè)計合成了6種疏水鏈不對稱的陽離子表面活性劑GHCC12-14、GHCC12-16、 GHCC14-16、GHBC12-14、GHBC12-16和GHBC14-16。通過1H-1H相關(guān)譜確定了長鏈異氰酸酯連接在N,N-二甲基氨基-1,2-丙二醇的端位羥基上；所有的中間體和目標(biāo)產(chǎn)物都通過MS、1H NMR和1H-1H gCOSY表征,結(jié)構(gòu)正確。考察了所合成的不對稱陽離子表面活性劑的γcmc、cmc、HLB值和臨界排列參數(shù)P等性質(zhì)。發(fā)現(xiàn)隨著碳鏈長度的增加表面張力和cmc增大,其中碳鏈長度為12-14的表面活性劑的γcmc和cmc都較小,具有較好的表面活性。將6種對稱型的陽離子表面活性劑和六種不對稱型的陽離子表面活性劑均與陰離子表面活性劑(SDBS、SDS和SAS)進(jìn)行復(fù)合制備囊泡,對其進(jìn)行外貌形態(tài)、粒徑、Zeta電位、電導(dǎo)率和透射電子顯微鏡TEM表征,發(fā)現(xiàn)陰-陽離子表面活性劑復(fù)合體系不論是富陽離子區(qū)還是富陰離子區(qū)均能形成囊泡。除X1=0.5時會出現(xiàn)沉淀或聚沉,粒徑基本都分布200～400 nm之間符合囊泡的范圍；Zeta電位在富陰離子區(qū)小于-30 mV,在富陽離子區(qū)大于30 mV,說明混合溶液自發(fā)形成的囊泡穩(wěn)定性較好。通過Masaniko幾何模型和Israelachvili理論對囊泡的形成機(jī)理進(jìn)行了闡述。將6種疏水鏈不對稱型陽離子表面活性劑進(jìn)行了生物學(xué)實驗研究。TEM表明,除GHBC14-16以外,不對稱陽離子類脂GHCn-m能夠較好地形成脂質(zhì)體；不對稱陽離子脂質(zhì)體與DNA復(fù)合后的粒徑和Zeta電位的研究表明,其粒徑大多在300-400 nm之間,符合細(xì)胞轉(zhuǎn)染的要求；瓊脂糖凝膠電泳實驗結(jié)果顯示,不對稱陽離子脂質(zhì)體GHCn-m與DNA的比例大于4：1時,對DNA具有較好的阻滯；當(dāng)GHCn-m/DNA為1：1時,不對稱陽離子脂質(zhì)體對Hela細(xì)胞的轉(zhuǎn)染效率的最高,比商品試劑DOTAP略高；對細(xì)胞毒性進(jìn)行測試,發(fā)現(xiàn)不對稱脂質(zhì)體對Hela細(xì)胞的毒性較大。
[Abstract]:In recent years, the research of gene therapy has been one of the most important fields in the medical field. It has been used in clinic for tumor, immunodeficiency and other diseases, but its development is seriously affected by its high toxicity and relatively low efficiency. Compared with the low safety of viral vectors, the low immune response, low toxicity and easy preparation of non-viral vectors make it a hot research topic. In this paper, six kinds of cationic surfactants, GHCC12-14, GHCC14-16, GHBC12-14, GHBC12-16 and GHBC14-16, have been designed and synthesized. The long chain isocyanate was linked to the terminal hydroxyl of N (N) -dimethyl amino-1o 2-propanediol by 1H-1H correlation spectroscopy, and all the intermediates and target products were characterized by MS ~ (1H) NMR and 1H-1H gCOSY, and the structure was correct. The HLB values and critical permutation parameters P of the synthesized asymmetric cationic surfactants were investigated. It is found that the surface tension and cmc increase with the increase of carbon chain length. The 緯 cmc and cmc of surfactant with carbon chain length 12-14 are smaller and have better surface activity. Six kinds of symmetric cationic surfactants and six asymmetric cationic surfactants were combined with anionic surfactants SDBSU SDS and SAS to prepare vesicles, and their appearance, size and Zeta potential were analyzed. The results of conductivity and transmission electron microscopy (TEM) TEM show that vesicles can be formed in anion or anionic regions of anion / cationic surfactant composite system. Except for X _ 1 = 0.5, precipitation or sedimentation would occur, and the particle size distribution was almost 200 ~ 400 nm. The Zeta potential was less than -30 MV in anion rich region and more than 30 MV in Fuyang ion region, which indicated that the vesicle formed spontaneously in mixed solution had better stability. The mechanism of vesicle formation was explained by Masaniko geometric model and Israelachvili theory. Six kinds of asymmetric cationic surfactants with hydrophobic chain were studied. Tem showed that GHCn-m with asymmetric cationic liposomes could form liposomes except GHBC14-16. The study on the particle size and Zeta potential of asymmetric cationic liposomes combined with DNA showed that most of them were between 300nm and 400nm, which met the requirements of cell transfection. When the ratio of asymmetric cationic liposome GHCn-m to DNA was greater than 4:1, the inhibition of DNA was better. When GHCn-m/DNA was 1:1, the transfection efficiency of asymmetric cationic liposome to Hela cells was the highest, which was slightly higher than that of commercial reagent DOTAP. It was found that asymmetric liposomes were more toxic to Hela cells.
【學(xué)位授予單位】：大連理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R450;O647.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 陳彥祥;喬衛(wèi)紅;劉棟良;李宗石;;陽離子脂質(zhì)體的轉(zhuǎn)染機(jī)制及轉(zhuǎn)染效率影響因素[J];生物工程學(xué)報;2007年05期

，

本文編號：1784239