9型重组腺相关病毒转染microRNA-21前体在大鼠心肌的转染效率及安全性

发布时间：2018-04-23 23:19

本文选题：腺相关病毒 + 型重组腺相关病毒　；参考：《中国现代医学杂志》2013年09期

【摘要】：目的研究含荧光标记ZsGreen的9型重组腺相关病毒(rAAV9)转染目的基因microRNA-21前体(pre-miR-21)至大鼠心肌的转染效率、表达时间及安全性。方法用冠脉注射方法转染PBS及rAAV9-Zs-Green-pre-miR-21 1.0×1010vg/只、1.0×1011vg/只、1.0×1012vg/只至SD大鼠心肌,倒置荧光显微镜观察心肌组织荧光表达并计算转染效率,Realtime PCR方法测定miR-21表达,超声心动图观察大鼠心功能。结果冠脉注射后7 d PBS组未见绿色荧光,余各组大鼠心肌均可观察到少量绿色荧光表达,第14天达高峰,之后呈下降趋势。14 d高病毒量组较其他中、低病毒量组转染效率明显高([73.7±8.4)%vs(39.6±4.8)%,(P0.001)];([73.7±8.4)%vs(24.1±4.2)%,(P0.001)]。14 d miR-21表达在低、中、高病毒量组较PBS组分别增加1.51倍、2.89倍及4.43倍。rAAV9转染后对大鼠心功能无影响(P0.05)。结论 rAAV9-Zs-Green-pre-miR-21可以有效、持久、安全地在大鼠心肌表达。
[Abstract]:Objective to study the transfection efficiency, expression time and safety of recombinant adeno-associated virus type 9 (rAAV9) containing fluorescent labeled ZsGreen into rat myocardium after transfection of microRNA-21 precursor pre-miR-21). Methods Coronary artery injection was used to transfect PBS and rAAV9-Zs-Green-pre-miR-21 1.0 脳 1010vg/ and 1.0 脳 1011vg/ to SD rat myocardium. The fluorescence expression of myocardial tissue was observed by inverted fluorescence microscope and the expression of miR-21 was measured by Realtime PCR method. The cardiac function of rats was observed by echocardiography. Results there was no green fluorescence in the PBS group 7 days after coronary artery injection. A small amount of green fluorescence expression was observed in the myocardium of the rest of the rats, which reached the peak on the 14th day, and then decreased. The amount of high virus in the 14 days group was higher than that in the other groups. The transfection efficiency ([73.7 卤8.4)%vs(39.6 卤4.8 卤P0.001)] ([73.7 卤8.4)%vs(24.1 卤4.2 + P0.001] 路14 d miR-21 expression in the low viral group was significantly higher than that in the PBS group. The expression of miR-21 in the high virus group was 1.51 times, 2.89 times and 4.43 times higher than that in the PBS group, respectively. Conclusion rAAV9-Zs-Green-pre-miR-21 can be expressed effectively, persistently and safely in rat myocardium.
【作者单位】：中南大学湘雅三医院心内科;广州医学院第三附属医院心内科;
【分类号】：R346

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