重组9型腺相关病毒载体转染小鼠心脏及对心功能的影响  

Transfection of recombinant adeno-associated virus serotype 9 to mouse heart in vivo and the impact on cardiac function

作　　者：向阳[1] 马依彤[1] 杨毅宁[1] 陈邦党[1] 刘芬[1] 高霞[1] 杜雷[1] 苗娜[2] 

机构地区：[1]新疆医科大学第一附属医院心脏中心,乌鲁木齐830054 [2]新疆医科大学第一附属医院病理科,乌鲁木齐830054

出　　处：《中华微生物学和免疫学杂志》2009年第10期899-902,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金项目（30760263）;新疆自治区卫生厅青年科技人才专项基金（2007Y23）

摘　　要：目的探讨重组9型腺相关病毒（recombinant adeno—associated serotype9，rAAV9）载体对小鼠心脏的转染效果及对心功能的影响。方法16只昆明种小鼠经尾静脉将携带增强型绿色荧光蛋白（enhanced green fluorescent protein，eGFP）报告基因的rAAV9（rAAV9-eGFP）转染入小鼠，于7、14、21、28d留取标本，使用荧光显微镜观察eGFP在心肌、肝、肺、肾、脑、骨骼肌的荧光表达，Westem blot法检测eGFP的蛋白表达。另20只昆明种小鼠随机分为2组，每组10只，分别经尾静脉注射rAAV9-eGFP和生理盐水，28d后行心脏超声及血流动力学检查。结果rAAV9-eGFP经尾静脉注射21d时eGFP表达在心脏达到高峰，转染效率可达32％，在其他脏器表达量少或无表达。rAAV9-eGFP组较生理盐水组心功能差异无统计学意义（P〉0．05）。结论rAAV9可通过外周静脉注射在心脏较高表达，且对心功能无不良影响。Objective To evaluate the transfection efficiency of recombinant adeno associated virus serotype 9 carrying enhanced green fluorescent protein （rAAV9-eGFP） to mouse heart in vivo and the impact on cardiac function. Methods Sixteen Kunming（KM） mice were transfected rAAV9-eGFP by tail injection, eGFP expression in the heart, liver, lung, kidney and brain cryosections was observed under inverted fluorescence microscope 7, 14, 21,28 days after the injection of rAAV9-eGFP and eGFP was quantitated by Western blot. Twenty KM mice were divided into control group and rAAV9-eGFP group randomly, and were received with saline or rAAV9-eGFP. The echocardiography and hemodynamics were performed 28 days after the injection of saline or rAAV9-eGFP. Results eGFP expression in the heart reached the maximum at day 21, at the point of which the transduction efficiency of rAAV9-eGFP in myoeardium was 32%. The other tissues had a little or no eGFP expression. The cardiac function did not reveal significant difference between rAAV9-eGFP group and the control group after transfection （ P 〉 0.05 ）. Conclusion rAAV9-eGFP gene can be stably and efficiently expressed in mouse heart, and has no toxic effect on cardiac function.

关 键 词：9型腺相关病毒 绿色荧光蛋白 转染 心功能 

分 类 号：R54[医药卫生—心血管疾病]