穿膜肽提高纳米银穿膜活性的研究.PDF

·论 著· 穿膜肽提高纳米银穿膜活性的研究 1,2 1,2 1 3 1* 2* 刘金华 ，郭倩倩 ，华海婴 ，袁波 ，赵永星 ，黄永焯 (1.郑州大学，郑州 450001 ；2. 中科院上海药物所，上海 201203 ； 3. 山东中医药大学附属医院，济南 250000) 摘要：目的 合成纳米银，在其表面修饰穿膜肽(TAT) ，并检测修饰后纳米银粒对人乳腺癌耐阿霉素细胞(MCF-7/ADR) 的穿膜活性。方法 通过化学还原法制备纳米银(AgNP) ，并通过Ag-S 共价键与 TAT 连接修饰 AgNP 。通过粒度仪、透 射电镜、激光共聚焦显微镜、流式细胞仪以及二喹林甲酸法等仪器及方法对其表征、共价连接及TAT 的介导活性进行测 定。结果 成功制备了 10 nm 以下的AgNP ，且修饰TAT 后的AgNP(AgNP-TAT)表现出了比AgNP 更强的穿膜活性。结 论 TAT 修饰AgNP 后能显著提高其穿膜活性。 关键词：纳米银；穿膜肽；MCF-7/ADR ；细胞摄取 中图分类号：R966 文献标志码：A 文章编号：1007-7693(2012)10-0865-04 The Penetrating Activity of Nano-silver Mediated by Cell Penetrating Peptide LIU Jinhua1,2 1,2 1 3 1* 2* , GUO Qianqian , HUA Haiying , YUAN Bo , ZHAO Yongxing , HUANG Yongzhuo (1.Zhengzhou University, Zhengzhou 450001, China; 2.Shanghai Institute of Materia Medica Chinese Academy of Sciences, Shanghai 201203, China; 3.Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Ji’nan 250000, China) ABSTRACT: OBJECTIVE To assess the intracellular delivery efficiency of silver nanoparticles surface-modified by cell penetrating peptides(TAT) in human breast cancer drug doxorubicin-resistant MCF-7/ADR cells. METHODS Silver nanoparticle(AgNP) was prepared by chemical reduction and was modified with TAT by Ag-S covalent bond. The TAT-AgNP was characterized using particle analyzer, TEM, laser confocal microsopy, and TGA, and TAT-mediated cellular uptake were measured using FACS. RESULTS The AgNP with diameter below 10 nm was successfully synthesized. The cell penetrating activity of TAT-modified AgNP (AgNP-TAT) was higher than that of AgNP. CONCLUSION The cell penetrating activity of AgNP was significantly improved by modification with TAT. KEY WORDS: silver nanopar