【摘要】：背景:人工支撐脂雙層膜是最接近細胞膜的體外仿生模型,構(gòu)建結(jié)構(gòu)與功能均類似于真實細胞膜的脂雙層膜具有十分重要的意義。目的:探索表面均勻且流動性高的支撐脂雙層膜的最佳制備條件并檢測指標。方法:采用薄膜擠壓法,分別以孔徑為0.1,0.2μm的濾膜對脂質(zhì)體分子過濾10次,保留未過濾樣品作為對照組,通過原子力顯微鏡對其表面粗糙程度進行檢測,探討制備均勻程度較高的脂雙層膜所需濾膜的最佳孔徑。將160μL卵磷脂(10 g/L)分別與5,10,25μL熒光磷脂(1 g/L)進行混合,通過熒光漂白恢復(fù)技術(shù)對脂雙層膜的流動性進行檢測,探討卵磷脂與熒光磷脂的最適比例。結(jié)果與結(jié)論:原子力顯微鏡實驗結(jié)果顯示,經(jīng)過孔徑為0.1μm濾膜過濾所得的脂雙層膜表面均勻程度最高(P0.01),其表面均方根粗糙度為(0.432±0.181)nm,當濾膜孔徑大于0.1μm時,脂雙層膜表面均勻程度差且存在較多的未鋪展開脂質(zhì)體囊泡。熒光漂白恢復(fù)實驗結(jié)果顯示,采用孔徑為0.1μm濾膜過濾的脂雙層膜熒光恢復(fù)程度最高,當卵磷脂(10 g/L)與熒光磷脂(1 g/L)體積比為160∶10時,采用孔徑為0.1μm濾膜過濾的脂雙層膜熒光恢復(fù)程度達到90%,且擴散系數(shù)大于1μm~2/s,符合優(yōu)質(zhì)脂雙層膜的標準。實驗提示利用薄膜擠壓法制備脂雙層膜時,卵磷脂與熒光磷脂的最適比例以及最佳濾膜孔徑。
[Abstract]:Background: artificially supported lipid bilayer membrane is the closest biomimetic model in vitro. It is very important to construct lipid bilayer membrane with structure and function similar to that of real cell membrane. Objective: to explore the optimum preparation conditions and detection index of the lipid bilayer membrane with uniform surface and high fluidity. Methods: the surface roughness of liposomes was determined by atomic force microscope (AFM). The liposome molecules were filtered 10 times by the filter membrane with the pore diameter of 0.1 渭 m and the unfiltered samples were retained as the control group by the thin film extrusion method, and the surface roughness of the liposomes was determined by atomic force microscope (AFM). The optimum pore size of lipid bilayer membrane with high homogeneity was discussed. 160 渭 L lecithin (10 g / L) was mixed with 5 10 渭 L fluorescent phospholipid (1 g / L). The fluidity of lipid bilayer membrane was detected by fluorescence bleaching recovery technique, and the optimum ratio of lecithin to fluorescent phospholipid was discussed. Results and conclusion: the results of atomic force microscope (AFM) showed that the surface uniformity of lipid bilayer membrane was the highest (P0.01), and the RMS roughness was (0.432 鹵0.181) nm. When the pore diameter of the membrane was greater than 0.1 渭 m, the surface homogeneity of lipid bilayer membrane was the highest (P0.01), and the mean square roughness of the membrane was (0.432 鹵0.181) nm. The surface uniformity of lipid bilayer membrane was poor and there were more unspread liposome vesicles. The results of fluorescence bleaching recovery experiment showed that the fluorescence recovery of lipid bilayer membrane was the highest when the volume ratio of lecithin (10 g / L) to fluorescent phospholipid (1 g / L) was 160: 10. The fluorescence recovery degree of lipid bilayer membrane filtered with 0.1 渭 m pore diameter was 90%, and the diffusion coefficient was more than 1 渭 m ~ 2 / s, which met the standard of high quality lipid bilayer membrane. It is suggested that the optimum ratio of lecithin to fluorescent phospholipids and the optimum pore size of the membrane can be obtained when the bilayer lipids are prepared by thin-film extrusion.
【作者單位】： 華南理工大學生物科學與工程學院;
【基金】：國家自然科學基金(11672109,11272125,11432006)~~
【分類號】：R318.08
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本文編號：2143856