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含二硫键类蛋白质脂质体的制备及其还原响应释放特性

Preparation of liposome bearing disulfide proteinoid and its reduction-responsive release property.

作者信息

Kwon Kyeongnan, Kim Jin-Chul

机构信息

a Department of Medical Biomaterials Engineering, College of Biomedical Science and Institute of Bioscience and Biotechnology , Kangwon National University , Chuncheon , Republic of Korea.

出版信息

J Biomater Sci Polym Ed. 2017 Sep;28(13):1365-1381. doi: 10.1080/09205063.2017.1328729. Epub 2017 May 18.

Abstract

Egg phosphatidylcholine (Egg PC) liposome bearing a disulfide proteinoid exhibited a reduction-responsive release property. Proteinoid composed of Asp, Leu, and cystamine (Prot(ALC)) and Asp and Leu (Prot(AL)) were synthesized by thermal condensation, confirmed by Raman, FT-IR, H NMR and C NMR spectroscopy. Egg PC liposome bearing the proteinoid was prepared by a film hydration and sonication method. The fluorescence quenching of dye (i.e. calcein) loaded in liposome bearing the proteinoid was 68.1-78.1%. The mean hydrodynamic diameter of liposome bearing the proteinoid was less than 200 nm and it decreased with increasing amount of the proteinoid. On the TEM photo, multi-lamellar vesicles were observed and the vesicle diameter was 100-300 nm. At all the phospholipid to proteinod ratios tested (i.e. 1:0.01, 1:0.02, and 1:0.05 (w/w)), the release degree in 12 h of dye loaded in liposome bearing Prot(AL) was less than 2%, and it was almost the same regardless of dithiothreitol (DTT, a reducing agent) concentration (i.e. 0, 10, and 20 mM). Whereas the release degree of dye loaded in liposome bearing Prot(ALC) was 1.5-8.4% and it was significantly dependent on DTT concentration.

摘要

携带二硫键类蛋白的鸡蛋磷脂酰胆碱(Egg PC)脂质体表现出还原响应释放特性。由天冬氨酸、亮氨酸和胱胺组成的类蛋白(Prot(ALC))以及天冬氨酸和亮氨酸(Prot(AL))通过热缩合合成,经拉曼光谱、傅里叶变换红外光谱、氢核磁共振光谱和碳核磁共振光谱确认。通过薄膜水化和超声处理方法制备了携带类蛋白的Egg PC脂质体。负载于携带类蛋白的脂质体中的染料(即钙黄绿素)的荧光猝灭率为68.1 - 78.1%。携带类蛋白的脂质体的平均流体动力学直径小于200 nm,并且随着类蛋白量的增加而减小。在透射电子显微镜照片上,观察到多层囊泡,囊泡直径为100 - 300 nm。在所有测试的磷脂与类蛋白比例(即1:0.01、1:0.02和1:0.05(w/w))下,负载Prot(AL)的脂质体中染料在12小时内的释放度小于2%,并且无论二硫苏糖醇(DTT,一种还原剂)浓度(即0、10和20 mM)如何,释放度几乎相同。而负载Prot(ALC)的脂质体中染料的释放度为1.5 - 8.4%,并且显著依赖于DTT浓度。

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