普朗尼克—聚乙烯亚胺纳米胶束的制备及其细胞生物学特性

doi:10.3785/j.issn.1008-9292.2017.04.03

浙江大学学报（医学版）

2017, Vol. 46

Issue (2): 127-133 DOI: 10.3785/j.issn.1008-9292.2017.04.03

专题报道

普朗尼克—聚乙烯亚胺纳米胶束的制备及其细胞生物学特性

汪河滨1,2(

),李洋2,刘馨刚2,周峻2,王青青2,汤谷平2,*(

)

1. 新疆兵团南疆化工资源利用工程实验室塔里木大学生命科学学院, 新疆阿拉尔 843300
2. 浙江大学化学系, 浙江杭州 310058

Preparation, characterization and cytology study of Pluronic-PEI micelles

WANG Hebin1,2(

),LI Yang2,LIU Xingang2,ZHOU Jun2,WANG Qingqing2,TANG Guping2,*(

)

1. Engineering Laboratory of Chemical Resources Utilization in South Xinjiang of Xinjiang Production and Construction Corps, College of Life Science, Tarim University, Xinjiang Alar 843300, China
2. Department of Chemistry, Zhejiang University, Hangzhou 310027, China

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摘要：

目的：采用普朗尼克—聚乙烯亚胺纳米胶束制备高效、低毒的药物和基因输送体系。方法：利用小分子量的聚乙烯亚胺作为交联剂制备普朗尼克—聚乙烯亚胺纳米胶束，采用动态光散射法、芘荧光探针法分别测定聚合物的粒径、表面电动电位及临界胶束浓度，考察聚合物携带基因和药物的能力，并在细胞水平上探讨普朗尼克—聚乙烯亚胺的细胞毒性、转染效率以及对细胞内ATP和P-糖蛋白水平的影响。结果：成功制备了普朗尼克—聚乙烯亚胺纳米胶束，其粒径120~180 nm，表面电位+6~+9 mV，具有良好的携载基因、药物的能力。细胞生物学研究表明，普朗尼克—聚乙烯亚胺毒性较小，其中P123-PEI600基因转染效率高，并且能降低细胞内ATP和P糖蛋白水平。结论：普朗尼克—聚乙烯亚胺是一种良好的药物和基因输送体系，P123-PEI600是理想的可用于逆转肿瘤多药耐药的载体。

关键词： 工艺学，制药; 聚乙烯亚胺/化学合成; 纳米球; 药物载体; 基因; 转染; 腺苷三磷酸; P-糖蛋白; 抗药性，肿瘤

Abstract:

Objective: To prepare and characterize Pluronic-PEI micelles as a drug/gene delivery system. Methods: We used the low-molecular-weight PEI as a cross-linking agent to prepare the Pluronic-PEI micelles. The particle size, zeta potential and critical micelle concentration (CMC) were measured by dynamic light scattering (DLS) and pyrene fluorescence probe. The cytotoxicity, transfection efficiency and the impact on the intracellular ATP and P-gp levels of Pluronic-PEI micelles were investigated at the cellular level. Results: Pluronic-PEI micelles were successfully prepared with a suitable particle size (120-180 nm), zeta potential (+6-+9 mv), and a good ability to carry the drug/gene. An in-vitro study showed that Pluronic-PEI had low cytotoxicity, and the P123-PEI600 possessed high gene transfection efficiency and could downregulate the intracellular ATP and P-gp levels. Conclusion: Pluronic-PEI is a good drug/gene delivery system, and P123-PEI600 is an ideal vector, which may be used in the combination therapy for reversing multidrug resistance.

Key words: Technology, pharmaceutical Polyethyleneimine/chemical synthesis Nanospheres Drug carriers Genes Transfection Adenosine triphosphate P-glycoprotein Drug resistance, neoplasm

收稿日期: 2016-11-02 出版日期: 2017-08-07

基金资助: 国家自然科学基金（51573161）

通讯作者: 汤谷平 E-mail: wanghebin329@163.com;tangguping@zju.edu.cn

作者简介: 汪河滨(1980—)，男，博士，主要从事生物医用纳米材料方面的研究；E-mail：wanghebin329@163.com；http://orcid.org/0000-0003-2159-914X

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引用本文:

汪河滨,李洋,刘馨刚,周峻,王青青,汤谷平. 普朗尼克—聚乙烯亚胺纳米胶束的制备及其细胞生物学特性[J]. 浙江大学学报（医学版）, 2017, 46(2): 127-133.

WANG Hebin,LI Yang,LIU Xingang,ZHOU Jun,WANG Qingqing,TANG Guping. Preparation, characterization and cytology study of Pluronic-PEI micelles. J Zhejiang Univ (Med Sci), 2017, 46(2): 127-133.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2017.04.03 或 http://www.zjujournals.com/xueshu/med/CN/Y2017/V46/I2/127

图 1 普朗尼克—聚乙烯亚胺的合成路线

图 2 紫杉醇吸收曲线和标准曲线

图 3 普朗尼克—聚乙烯亚胺在重水中的核磁共振氢谱图

表 1 普朗尼克—聚乙烯亚胺元素分析结果

图 4 不同普朗尼克—聚乙烯亚胺空白胶束的透射电镜图

表 2 普朗尼克—聚乙烯亚胺粒径和电位分析结果

图 5 普朗尼克—聚乙烯亚胺溶液浓度对数值随荧光强度比值变化曲线图

图 6 普朗尼克—聚乙烯亚胺的DNA凝胶电泳图

表 3 普朗尼克—聚乙烯亚胺载药率和包封率

图 7 不同普朗尼克—聚乙烯亚胺的药物释放曲线

图 8 加入普朗尼克—聚乙烯亚胺的细胞生存活曲线

图 9 不同普朗尼克—聚乙烯亚胺和PEI25 000携带pEGFP质粒在HEK293和A549细胞上的转染结果

图 10 普朗尼克—聚乙烯亚胺对细胞内ATP和P-gp水平的影响

图 11 普朗尼克—聚乙烯亚胺对Panc-1细胞内P-gp的影响

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