重组腺病毒miR-15b对心肌细胞凋亡的影响  

Effect of recombinant adenoviral miR-15b on apoptosis of cardiomyocytes

作　　者：刘丽凤[1] 刘秀华[2] 白静[1] 陈杰[1] 王一茹[1] 王禹[1] 

机构地区：[1]解放军总医院老年心血管病研究所,北京100853 [2]解放军总医院病理生理研究室,北京100853

出　　处：《中华老年心脑血管病杂志》2012年第5期535-537,共3页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

摘　　要：目的通过构建miR-15b过表达及低表达腺病毒载体,探索其对心肌细胞凋亡的影响。方法体外分离培养SD乳鼠原代心肌细胞,建立模拟心肌缺氧/复氧(H/R)模型。实验随机分为对照组、H/R组、miR-15b过表达组(AdmiR-15b组)、miR-15b低表达组(AdasmiR-15b组)、miR-15b阴性对照组(AdmiR-NC组)。Annexin V-异硫氰酸荧光素标记/碘化丙啶标记细胞后,流式细胞仪检测细胞凋亡。Taqman实时定量PCR法精确定量miR-15b表达水平。结果细胞转染效率为100%。与AdmiR-NC组比较,AdmiR-15b组miR-15b表达上调>9倍,而AdasmiR-15b组表达下调50%(P<0.01)。与对照组比较,H/R组、AdmiR-NC组细胞凋亡率明显升高(P<0.01);与AdmiR-NC组比较,AdmiR-15b组细胞凋亡率明显升高,AdasmiR-15b组明显降低(P<0.01)。结论在心肌细胞miR-15b为重要的凋亡调节因子,阻断miR-15b表达有可能减少心肌细胞凋亡,从而保护心肌减轻再灌注损伤。Objective To study the effect of recombinant adenoviral microRNA-15b on apoptosis of cardiomyocytes by constructing its over-expression and low-expression vectors.Methods Primary myocardiocytes of neonatal SD rats were isolated in vitro.A myocardial hypoxia/reoxygenration（H/R） model was established and exposed to hypoxia for 6 h and to reoxygenation for 24 h.The rats were divided into miR-15b overexpression（AdmiR-15b） group,low miR-15b expression（AdasmiR-15b） group,and miR negative control（AdmiR-NC） group.Apoptosis of myocardiomyocytes was detected by flow cytometry after stained with annexinⅤfluorescein isothiocyanate.Expression level of miR-15b was measured by Taqman quantitative RT-PCR.Results The transfection rate of myocardiocytes was 100%.The miR-15b expression level was over 9-fold higner in AdmiR-15b group than in AdmiR-NC group and 50% lower in AdasmiR-15b group than in AdmiR-NC group（P〈0.01）.The apoptosis rate of myocardiocytes was significantly higher in H/R group and AdmiR-NC group than in control group and in AdmiR-15b group than in AdmiR-NC group（P〈0.01）.Conclusion The miR-15b is an important factor for the apoptosis of myocardiomyocytes and down-regulation of its expression can reduce apoptosis of myocardiomyocytes,thus protecting myocardium against reperfusion injury.

关 键 词：细胞凋亡 腺病毒科 微RNAS 聚合酶链反应 转染 心肌再灌注损伤 

分 类 号：R363[医药卫生—病理学]