小鼠尾静脉注射与心肌注射腺病毒载体转染效率的比较

Comparison of the efficiency of tail vein injection and intramyocardial injection of adenovirus vector in mice

目的比较小鼠尾静脉注射或心肌注射腺病毒载体后心脏组织和肝脏组织中靶基因的转染效率。方法构建表达绿色荧光蛋白GFP的腺病毒载体(GFP-Ad)。C57BL/6小鼠20只随机分为尾静脉注射腺病毒载体组与心肌注射腺病毒载体组各10只,用实时定量PCR检测不同时间点小鼠心肌组织和肝脏组织中GFP的mRNA表达水平,并且通过荧光显微镜观察GFP荧光表达情况。结果心肌注射腺病毒载体组心脏组织中GFP的mRNA表达水平明显高于尾静脉注射腺病毒载体组,心肌注射腺病毒载体组心脏组织中荧光强度明显增强。同时,我们发现两组的肝脏组织中GFP的mRNA表达水平和荧光强度均明显高于心脏组织中;且在尾静脉注射腺病毒载体组,肝脏组织中GFP的mRNA表达水平和荧光强度在7 d达高峰,而心肌注射腺病毒载体组则在3 d达高峰。结论提高心脏组织中靶基因的转染效率宜采用心肌注射腺病毒载体的方法;对于肝脏组织转染效率,两种注射方法均可,鉴于尾静脉注射腺病毒载体的方法创伤小,宜采用。

Objective To investigate the efficiency of target gene transfection of the heart and liver after tail vein or intramyocardial injection of adenovirus vector（ GFP-Ad）. Methods GFP-AD was constructed at first. A total of 20 male 8-week old C57 BL /6 mice were randomly and equally divided into tail vein injection of GFP-AD group and intramyocardial injection of GFP-AD group. The mRNA levels of GFP in the heart and liver tissues were detected by Q-PCR at different time points. Fluorescence microscopy was performed to visualize the expression of GFP fluorescence. Results Compared with the tail vein injection group,the GFP mRNA level in mouse heart tissue was apparently higher in the intramyocardial injection group. In both groups,the GFP mRNA levels in liver tissue were significantly increased compared with that in the heart tissue. In the tail vein injection group,the GFP mRNA level in liver tissue reached a peak on day 7;but in the intramyocardial injection group,the mRNA level of GFP in liver tissue reached apeak on day 3. We also observed the same trend of GFP fluorescence expression in the tail vein injection group compared with that in theintramyocardial injection group. Conclusions Intramyocardial injection of adenovirus vector is suitable to achieve a higher transfection efficiency in mouse heart tissue compared with the tail vein injection method. Although both injection methods are suitable for transfection of mouse liver,the tail vein injection method is preferential for it is simple and less invasive.