重组1和2型腺相关病毒载体在小鼠心脏转导研究

Efficacy of transduction of recombinant adeno-associated virus vectors serotypes 1 and 2 to mouse heart

目的探讨重组1和2型腺相关病毒(rAAV1和rAAV2)载体向小鼠心脏转导效果。方法14只昆明小鼠应用经腹心包腔内注射术,分别将携带增强绿色荧光蛋白(EGFP)、荧光素酶(LUC)报告基因的rAAV1(rAAV1-EGFP组6只,rAAV1-LUC组1只)r、AAV2(rAAV2-EGFP组6只,rAAV2-LUC组1只)分别转导入小鼠的心包腔内,使用共聚焦显微镜观察EGFP在心肌的表达,冷光CCD相机体内监测LUC在心肌表达。结果rAAV1-EGFP组绿色荧光出现于心肌组织全层;rAAV2-EGFP组绿色荧光仅局限于心外膜下少数心肌细胞。rAAV1-EGFP组小鼠心肌细胞的表达范围和强度优于rAAV2-EGFP组。rAAV1-LUC组小鼠心肌组织表达强度、持续表达时间优于rAAV2-LUC组。结论在小鼠心脏中,rAAV1是一种比rAAV2更为有效的转基因载体。

Objective To determine the efficacy of tmnsduction of recombinant adeno-associated virus vectors semtypes 1 and 2 （rAAV1, rAAV2） to the mouse heart. Methods Recombinant adeno-assoeiated vires seretypes 1 and 2 carrying either enhanced green fluorescent protein（EGFP） or luciferase （LUC） reporter gene were separately injected into the pericardium of mice. After 30 days, cryosection was analyzed by confocal microsco- py to examine the expression of EGFP. Expression of LUC gene in myocardium was monitored with a cooled CCD camera on 14th and 45th days. Results Green fluorescence was detected in cryosection of the hearts in all mice. The expression of rAAV1-EGFP was seen in cardiac myocytes from epicardium to endoeardium. The expression of rAAV2-EGFP was limited in some cardiac myoeytes under epicardium. The intensity and range of rAAV1-EGFP expression were better than those of rAAV2. The expression intensity and duration of rAAV1-LUC reporter gene were better than those of rAAV2-LUC reporter gene in cardiac myoeytes. Conclusion As a gene-tmnsducing vector in mouse heart, rAAV1 is superior to rAAV2.