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| 胆甾醇修饰的PEG-P[Asp(DET)]/miRNA复合物的制备、理化性能及细胞摄取能力 |
| 马志强1,2,刘赣1,高红军1,吴姗2,贾婷婷2,钟延强2*,史林启1 |
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(1. 南开大学高分子化学研究所, 天津 300071;
2. 第二军医大学药学院药剂学教研室, 上海 200433
*通信作者) |
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| 摘要: |
| 目的 制备疏水基修饰的聚阳离子高分子基因载体PEG-P[Asp（DET）]-10%胆甾醇，对其复合miRNA的理化性质和细胞摄取能力进行研究。方法 开环聚合合成PEG-P[Asp（DET）]，再采用氯甲酸胆甾醇基进行疏水修饰，核磁共振法验证其结构；使其与hsa-miR-15a形成胶束复合物，对该胶束复合物的粒径、Zeta电位、稳定性、包封率以及细胞毒性进行考察；以白血病细胞K562细胞系为模型细胞进行体外细胞实验对其摄取进行考察。结果 合成的PEG-P[Asp（DET）]-10%胆甾醇具有良好的溶解性，可与miRNA形成稳定的胶束复合物，当氮磷比（N/P）=20，基因浓度在5 μmol/mL时，形成的胶束复合物粒径为（192.4±10.8）nm，Zeta电位为（6.9±0.9）mV，包封率为（90.5±3.2）%。该复合物在实验条件下表现出了相当的稳定性，细胞对该复合物的摄取能力明显强于商品化试剂脂质体lipo2000。结论 PEG-P[Asp（DET）]-10%胆甾醇是一种优良的高分子基因传递系统载体，在体外实验中可有效实现细胞对于miRNA 的稳定摄取。 |
| 关键词: 高分子胶束复合物 聚乙二醇-聚｛氮-[氮-(2-氨乙基)-2-氨乙基]天冬氨酸｝ 微mRNAs 基因传递系统 基因疗法 |
| DOI：10.3724/SP.J.1008.2014.00171 |
| 投稿时间：2013-07-15修订日期：2013-08-20 |
| 基金项目: |
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| Cholesterol modified PEG-P[Asp(DET)]/miRNA complex：preparation, physicochemical properties, and cellular uptake |
| MA Zhi-qiang1,2,LIU Gan1,GAO Hong-jun1,WU Shan2,JIA Ting-ting2,ZHONG Yan-qiang2*,SHI Lin-qi1 |
(1. Institute of Polymer Chemistry, Nankai University, Tianjin 300071, China;
2. Department of Pharmaceutics, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
*Corresponding author.) |
| Abstract: |
| Objective To synthesize PEG-P[Asp(DET)] with 10% cholesterol chloroformate modified on its side chain and to study the physicochemical properties and cellular uptake of polymer complex PEG-P[Asp(DET)]-chole/hsa-miR-15a. Methods PEG-P[Asp(DET)] was synthesized by ring opening polymerization and modification with cholesterol chloroformate to acquire hydrophobicity, and MRI was used to verify its structure. The particle size, Zeta potential, stability, encapsulation efficiency, and cytotoxicity of the polymer complex PEG-P[Asp(DET)]-10% chole/hsa-miR-15a were examined. Finally, in vitro cellular uptake experiment was carried out with the leukemia cell line K562. Results The synthesized polymer PEG-P[Asp(DET)]-10% chole had fine solubility and could form into stable polymer complex PEG-P[Asp(DET)]-chole/hsa-miR-15a. When nitrogen-phosphorus ratio (N/P) was at 20 and concentration of the miRNA was 5 μmol/mL, the particle size was (192.4±10.8) nm, Zeta potencial was (6.9±0.9) mV, and encapsulation efficiency was (90.5±3.2)%. The complex displayed good stability under experimental condition. In vitro cellular uptake experimental indicated that the uptake capacity of PEG-P[Asp(DET)]-10%chole/hsa-miR-15a was higher than that of the commercial agent lipo2000. Conclusion PEG-P[Asp(DET)]-10%chole is a fine gene polymer carrier for miRNA and can achieve stable cellular uptake of miRNA. |
| Key words: polymer complexes poly (ethylene glycol)-block-poly ｛N-[N-(2-aminoethyl)-2-a minoethyl] aspartamide｝ microRNAs gene delivery system gene therapy |
