阳离子双亲性多肽-脂质纳米粒子的构建及细胞毒性和体外稳定性

doi:DOi:10.12066/j.issn.1007-2861.1758

上海大学学报(自然科学版) ›› 2018, Vol. 24 ›› Issue (1): 142-150.doi: DOi:10.12066/j.issn.1007-2861.1758

• 研究论文 • 上一篇

阳离子双亲性多肽-脂质纳米粒子的构建及细胞毒性和体外稳定性

王乔¹^,², 费浩²()

^1. 上海大学生命科学学院, 上海 200444
^2. 中国科学院苏州纳米技术与纳米仿生研究所, 江苏苏州 215123

收稿日期:2016-03-16 出版日期:2018-02-28 发布日期:2018-03-05
通讯作者: 费浩 E-mail:hfei2008@sinano.ac.cn
基金资助:
国家自然科学基金资助项目(21302213);国家自然科学基金资助项目(81573339)

Construction of cationic amphipathic peptide-lipid nanoparticle and evaluation of its cytotoxicity and stability in vitro

WANG Qiao¹^,², FEI Hao²()

^1. School of Life Sciences, Shanghai University, Shanghai 200444, China
^2. Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences,Suzhou 215123, Jiangsu, China

Received:2016-03-16 Online:2018-02-28 Published:2018-03-05
Contact: FEI Hao E-mail:hfei2008@sinano.ac.cn

摘要/Abstract

摘要：

选用阳离子双亲性 R 多肽(RLARLLRRLARWLR)进行脂质纳米粒子的构建. 通过脂质乳浊液清除实验、色氨酸荧光光谱蓝移实验, 对 R 多肽与脂质的重组过程和作用方式进行表征. 通过透射电子显微镜观察、动态光散射、zeta 电位检测, 表征了所构建的阳离子双亲性多肽-脂质纳米粒子(R peptide-lipid nanoparticle, R-LNP)的粒径和表面性质. 通过 MTT 实验检测了 R-LNP 的细胞毒性; 通过超高效液相色谱(ultra high performance liquid chromatography, UPLC)方法比较了 R 多肽和 R-LNP 抗酶解(糜蛋白酶、胰蛋白酶)的稳定性. 结果表明, R 多肽能促进脂质的重组, 其疏水性部分嵌入脂质纳米粒子的疏水核心, 形成粒径为 (14.7$\pm $0.1) nm, zeta 电位为 17.8 mV 的脂质纳米粒子. R-LNP对A549 和 MCF-7 细胞的半抑制浓度为 (5.93$\pm $0.75) 和 (4.36$\pm $0.40) $\mu$mol/L, 保留 R 多肽细胞毒性作用, 同时 R-LNP 有效提高了 R 多肽的稳定性, 有助于多肽的体内应用.

关键词: 阳离子双亲性多肽, 脂质纳米粒子, 构建, 功能

Abstract:

In this paper, cationic amphipathic R peptide (RLARLLRRLARWLR) was chosen to construct peptide-lipid nanoparticle. Recombination and interaction of R peptide with lipid mixture were studied using lipid emulsion clearance assay and blue shift of tryptophan fluorescence assay. The diameter and surface characterization of cationic amphipathic R peptide-lipid nanoparticle (R-LNP) were obtained with transmission electron microscope (TEM), dynamic light scattering (DLS) and zeta potential measurements. MTT assay was used to investigate cytotoxicity of R-LNP. Stability of R-LNP and R peptide against chymotrypsin and trypsin was explored using ultra high performance liquid chromatography (UPLC). The results showed that R peptide caused reconstruction of lipid to form lipid nanoparticle with a diameter of (14.7$\pm $0.1) nm and zeta potential of 17.8 mV. Hydrophobic amino acids of R peptide inserted in the hydrophobic core of lipid nanoparticle. Cytotoxicity of R peptide was retained in R-LNP, of which half-inhibitory concentration for A549 and MCF-7 was (5.93$\pm $0.75) $\mu$mol/L and (4.36$\pm $0.40) $\mu$mol/L respectively. UPLC results showed that R-LNP increased stability of R peptide, facilitating its translational application in vivo.

Key words: cationic amphipathic peptide, lipid nanoparticle, construction, function

中图分类号:

王乔, 费浩. 阳离子双亲性多肽-脂质纳米粒子的构建及细胞毒性和体外稳定性[J]. 上海大学学报(自然科学版), 2018, 24(1): 142-150.

WANG Qiao, FEI Hao. Construction of cationic amphipathic peptide-lipid nanoparticle and evaluation of its cytotoxicity and stability in vitro[J]. Journal of Shanghai University（Natural Science Edition）, 2018, 24(1): 142-150.

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阳离子双亲性多肽-脂质纳米粒子的构建及细胞毒性和体外稳定性

Construction of cationic amphipathic peptide-lipid nanoparticle and evaluation of its cytotoxicity and stability in vitro

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