靶向Akt1基因RNA干扰对胸主动脉平滑肌细胞增殖的影响  

Effect of targeting Akt1 gene RNA intervention on proliferation of smooth muscle cell of rabbits

作　　者：刘苏健[1] 范波[2] 丁明超[1] 王意忠[1] 王斌[1] 

机构地区：[1]北京市航天中心医院外周血管介入科,100049 [2]河北省保定市第一医院

出　　处：《河北医药》2014年第18期2725-2727,共3页Hebei Medical Journal

摘　　要：目的构建兔Akt1短发卡RNA(shRNA)真核表达载体,探讨靶向Akt1基因RNA干扰对兔胸主动脉平滑肌细胞增殖的影响。方法经预实验挑选2条有效短发卡RNA转染入原代培养日本大耳白兔胸主动脉平滑肌细胞内,设置空白对照组(A组)、阴性对照组(B组)、Akt1-shRNA-1组(C组)、Akt1-shRna-2组(D组)。48、72 h荧光显微镜下观察细胞转染效率,分别在转染平滑肌细胞后24、48、72、96 h使用流式细胞仪和TUNEL法检测凋亡细胞数目。结果平滑肌细胞48 h转染效率为17.8%;流式细胞仪测定C组和D组在48 h凋亡率分别为19.72%和15.43%,抑制增值效果达到最高峰,与A、B组比较,差异有统计学意义(P<0.05)。TUNEL法测定结果表明,C、D组细胞凋亡水平与A、B组比较明显增多(P<0.05),且以C组较为明显。结论成功构建了2条兔Akt1 shRNA真核表达载体,可有效地抑制平滑肌细胞增殖。Objective To construct Akt1 shRNA eukaryon expression vector to investigate the effect of targeting Akt1 gene RNA intervention on proliferation of smooth muscle cell of rabbits. Methods After preliminary experiment,two strips of effective shRNA were selected to be transfected into thoracic aorta smooth muscle cells（ SMC） of rabbits（ Oryctolagus cuniculus）. The experiment was divided into four groups,blank control group（ group A）,negative control group（ group B）,Akt1-shRNA-1group（ group C）,Akt1-shRna-2 group（ group D）. The cell transfection rate was detected under fluorescence microscope at 48 h,72h after transfection,and the cell apoptosis rates were measured by flow cytometry and TUNEL respectively at 24 h,48h,72 h,96h after transfection. Results The two sequences of synthesized shRNA named Akt1-shRNA-1 and Akt1-shRNA-2 were exactly the same as the designed ones. The SMC transfection rate at 48 h was 17. 8%.The cell apoptosis rate was 19. 72% and 15. 43% at 48 h respectively in group C and Group D,there were significant differences between group C,group D and group A,group B（ P 〈0. 05）. The results by TUNEL showed that the levels of cell apoptosis in group C and group D were significantly increased,as compared with those in group A and group B（ P 〈0. 05）,especially in group C. Conclusion Two strips of Akt1 shRNA eukaryon expression vector in rabbits are successfully constructed,which can effectively inhibit the proliferation of smooth mouse cell of rabbits.

关 键 词：载体构建 AKT1 再狭窄 SHRNA 

分 类 号：R543.12[医药卫生—心血管疾病]