PreS1多肽(2-21Aa)介导肝脏靶向脂质体的制备及其靶向性研究  

The research of nano-liposome targeting hepatocyte mediated by PreS1 peptide (2-21Aa)

作　　者：韩超[1] 刘宏利[1] 李军[1] 蓝竹[1] 吴玉章[1] 韩俊峰[1] 

机构地区：[1]第三军医大学基础部免疫学教研室,重庆400038

出　　处：《免疫学杂志》2014年第3期240-245,共6页Immunological Journal

基　　金：重庆市自然科学基金(cstc2011jjA0624);国家自然科学基金(81170396);长江学者和创新团队发展计划资助项目(IRT1052)

摘　　要：目的以来源于乙肝病毒（hepatitisBvirus，HBV）PreSl抗原2-21Aa多肽（HBVpreS／2—21nyr）为导向的肝细胞靶向长效循环脂质体（10ngcirculatingliposome，LCL），通过体外细胞学实验验证其靶向性。方法固相合成HBVpreS／2—21m多肽，化学偶联Mal—PEG2000-DSPE（马来酰亚胺一聚乙二醇2000一硬脂酰基磷脂酰乙醇胺）生成HBVpreS／2五1q—PEG2000-DSPE；采用乙醇注入法制备脂质体，包裹荧光素钠（fluoresceinsodium，FS）形成HBVpreS／2—21nyt-FS—LCL；激光粒度仪、透射电镜、高效液相色谱检测和观察脂质体的粒径、超微形态及包封率；以稳定表达NTCP的HepG2为细胞模型，分析纳米脂质体靶向性。结果合成的靶向脂质体，呈类圆形，大小均匀，粒径为（94．09±9．2）nm，达到纳米级别；有效包裹荧光素钠，包封率为（89．32±1．02）％；与无靶向修饰脂质体相比，可递送更多的荧光素钠进入细胞内。结论HBVpreS／2—21myt修饰的靶向脂质体，有较高的包封率，为纳米脂质体，能特异性地通过NTCP靶向肝细胞，有望成为一种新型的肝细胞靶向给药系统。In this study, we aimed to establish a nano-liposome targeting hepatocytes via HBVpreS/2-21myr, which was derived from HBV preS1 antigen and could bind with HBV receptor, sodium taurocholate cotransporting polypeptide （NTCP） on the hepatocyte. To get the nano-liposomes, the peptide HBVpreS/2-21myr was synthesized by a solid-phase method, and coupled with Mal-PEGam-DSPE for constructing HBVpreS/2-21mry-PEGz0o0-DSPE as a targeted media, then taken HBVpreS/2-21myr-PEGz000-DSPE as one of components, the liposomes were made by the ethanol injection method. The liposome encapsulating fluorescein sodium （FS）, HBVpreS/2-21myr-FS-LCL, was produced and then its physical and chemical characters were detected using nanoparticle size analyzer, transmission electron microscope （TEM） and high performance liquid chromatography（HPLC）, respectively. HepG2-NTCP cell was built by transferring NTCP encoded plasmid to HepG2 cell line as a cell model for analyzing the target ability of the HBVpreS/2-21myr-FS-LCL. As a result, we found that the average particle sizes of the liposomes were （94.09±9.2） nm and dispersed evenly in the quasi-circular shape, consistent with what shown under the TEM, and the encapsulation efficiencies of the liposomes were （89.32±1.02）%. Compared to the liposomes （FS-LCL） without HBVpreS/2-21myr, the HBVpreS/2-21myr-FS-LCL could send more FS into HepG2-NTCP cells by a NTCP dependent manner. In conclusion, the HBVpreS/2-21myr-FS-LCL was a nano-liposome with a high encapsulation efficiency, and could specifically target the hepatocyte by the receptor NTCP, and it might be a novel drug delivery system for the therapy of liver diseases.

关 键 词：肝细胞靶向 纳米脂质体 钠离子-牛磺胆酸共转多肽 HBVpreS 2-21nyr 

分 类 号：R943[医药卫生—药剂学]