脂质体对细菌外膜囊泡的修饰及其体外评价

doi:10.5246/jcps.2019.08.055

中国药学(英文版) ›› 2019, Vol. 28 ›› Issue (8): 571-578.DOI: 10.5246/jcps.2019.08.055

脂质体对细菌外膜囊泡的修饰及其体外评价

王佳星¹, 王雨¹, 牟家慧², 魏潇萌², 殷梦雅¹, 刘雅婕¹, 李佳佳¹, 李馨儒^1*

1. 北京大学医学部药学院分子药剂学与新释药系统北京市重点实验室, 北京 100191
2. 中国医学科学院医药生物技术研究所, 北京 100050

收稿日期:2019-04-27 修回日期:2019-05-13 出版日期:2019-09-02 发布日期:2019-05-23
通讯作者: Tel.: +86-10-82801508, E-mail: ll@bjmu.edu.cn
基金资助:
National Natural Science Foundation of China (Grant No. 81573381) and CAMS Initiative for Innovative Medicine (Grant No. CAMS-I2M-1-012).

Decoration of bacterial outer membrane vesicles with liposomes

Jiaxing Wang¹, Yu Wang¹, Jiahui Mou², Xiaomeng Wei², Mengya Yin¹, Yajie Liu¹, Jiajia Li¹, Xinru Li^1*

1. Beijing Key Laboratory of Molecular Pharmaceutics and New Drug System; School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
2. Institute of Medicinal Biotechnology, Chinese Academy of Medical Science & Peking Union Medical College, Beijing 100050, China

Received:2019-04-27 Revised:2019-05-13 Online:2019-09-02 Published:2019-05-23
Contact: Tel.: +86-10-82801508, E-mail: ll@bjmu.edu.cn
Supported by:
National Natural Science Foundation of China (Grant No. 81573381) and CAMS Initiative for Innovative Medicine (Grant No. CAMS-I2M-1-012).

摘要/Abstract

摘要：

本研究使用脂质体对细菌外膜囊泡进行修饰, 并对修饰后的外膜囊泡进行体外评价。采用环丝氨酸诱导结合硫酸铵沉淀离心法提取绿脓杆菌外膜囊泡, 并对其粒径、分布、电荷、蛋白含量和稳定性进行测定。采用薄膜分散法制备若干种脂质体, 分别使用涡旋、超声和挤出法修饰膜囊泡并测定其粒径和电荷。采用荧光共振能量转移技术研究脂质体与膜囊泡相互作用。研究结果表明, 细菌膜囊泡为球形, 粒径在200 nm范围, 带负电荷。采用涡旋的方式对修饰后的膜囊泡的粒径与分布影响不大, 采用超声工艺, 使膜囊泡修饰后粒径和分布均下降, 而采用挤出工艺, 粒径与分布均增加。FRET实验预示脂质体与膜囊泡发生膜融合。上述结果表明脂质体可成功对膜囊泡进行修饰, 修饰后的膜囊泡展示出可调控的理化性质, 这给膜囊泡带来更多的应用。

关键词: 膜囊泡, 脂质体, 粒径, 荧光共振能量转移, 融合

Abstract:

In this study, liposomes were used to decorate bacterial outer membrane vesicles (OMVs), and decorated OMVs were evaluated in vitro. The OMVs of Pseudomonas aeruginosa were extracted by pressure-induced ammonium sulfate precipitation, and their particle size, distribution, zeta potential, protein content and stability were determined. Several types of liposomes were prepared by thin film dispersion method, and the OMVs were decorated by vortexing, sonication and extrusion, respectively. The interaction between liposome and OMV was studied with fluorescence resonance energy transfer (FRET) method. The results showed that the OMVs were spherical in shape and negatively charged. The vortexing method exerted little effect on the particle size and distribution of the decorated OMVs. The sonication process reduced the particle size and distribution of OMVs. FRET experiment indicated that the OMVs were decorated through membrane fusion. The above-mentioned results indicated that liposomes could successfully decorate OMVs, and decorated OMVs certainly widened their applications.

Key words: Outer membrane vesicle, Liposome, Particle size, FRET, Fusion

中图分类号:

R943

Supporting:

王佳星, 王雨, 牟家慧, 魏潇萌, 殷梦雅, 刘雅婕, 李佳佳, 李馨儒. 脂质体对细菌外膜囊泡的修饰及其体外评价[J]. 中国药学(英文版), 2019, 28(8): 571-578.

Jiaxing Wang, Yu Wang, Jiahui Mou, Xiaomeng Wei, Mengya Yin, Yajie Liu, Jiajia Li, Xinru Li. Decoration of bacterial outer membrane vesicles with liposomes[J]. Journal of Chinese Pharmaceutical Sciences, 2019, 28(8): 571-578.

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脂质体对细菌外膜囊泡的修饰及其体外评价

Decoration of bacterial outer membrane vesicles with liposomes

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