靶向HIV-1细胞受体基因CCR5的不对称siRNA的稳定性和体内分布研究  

Stability and in vivo Distribution of Asymmetric siRNA Targeting HIV-1 Coreceptors CCR5

作　　者：张昌栋[1] 殷妍[1] 郭佳佳[1] 梁晓飞[2] 段友容[2] 吴稚伟[3] 王建军[1] 徐根兴[3,4] 

机构地区：[1]南京大学生命科学院,南京210008 [2]上海市肿瘤研究所,上海200032 [3]南京大学医学院公共健康医学中心和生命分析化学国家重点实验室,南京210093 [4]江苏省基因药物工程技术研究中心,苏州215128

出　　处：《药学与临床研究》2011年第5期391-396,共6页Pharmaceutical and Clinical Research

基　　金：国家重大新药创制项目(2009ZX09103-635);国家自然科学基金(30870124);江苏省自然科学基金(BK2009109)

摘　　要：目的:探讨靶向HIV-1细胞受体基因CCR5的不对称siRNA(asiRNA)在血清和动物体内的稳定性和在动物体内分布情况。方法:用内皮抑素同脂质胶束联合递送asiRNA,琼脂糖凝胶电泳法检测asiRNA的血清稳定性,ELISA法和活体荧光成像法检测asiRNA的体内分布。结果:改变合适的实验条件和asiRNA的修饰,可提高asiRNA的血清稳定性。用ELISA法检测出静脉给药15 min后双链asiRNA在血液、肝、肺和肾内浓度较高,用活体荧光成像法检测出静脉给药24 h后asiRNA向肾内汇集排泄,两种检测方法测得asiRNA在脾内浓度较低。结论:两种检测方法方便实用,灵敏度不同,均可用于阳离子脂质体递送asiRNA的体内分布检测。Objective： To evaluate the stability in serum and distribution in vivo of asymmetric siRNA（asiRNA） targeting HIV-1 coreceptors CCR5.Methods： Agarose gel electrophoresis was used to detect the stability of asiRNA combined lipid micelles and endostatin in serum.ELISA-based assay and in vivo fluorescence imaging were used to detect the in vivo distribution of asiRNA.Results： Modification of asiRNA improved its stability in serum at appropriate conditions.Higher distribution of asiRNA occurred in blood,liver,lung and kidney at 15 min and the highest occurred in the kidney at 24 hour after administration,while lower levels of asiRNA occurred in spleen.Conclusion： Both methods are,with different sensitivities,convenient for detecting the distribution of asiRNA targeting CCR5 delivered via cationic liposome after intravenous administration,particularly from a pharmacokinetic perspective.

关 键 词：CCR5 SIRNA 不对称siRNA ELISA 活体荧光成像 

分 类 号：R318[医药卫生—生物医学工程] Q78[医药卫生—基础医学]