CAJ | 학술논문

目的观察RNA干扰(RNAi)沉默成年大鼠心肌细胞缺氧诱导因子-1α(HIF-1α)的实验可能性和转染效率.方法构建靶向HIF-1α小干扰RNA基因,并转染到成年大鼠细胞中.反转录-聚合酶链反应(RT-PCR)检测成年大鼠心肌细胞的HIF-1 αmRNA水平变化,噻唑蓝(MTT)法检测不同浓度的作用下大鼠心肌细胞活力,转染后24h内荧光显微镜检测转染对照的转染效果.结果转染小干扰RNA(siRNA)结果显示si-r-HIF-1α的沉默有效性达到73％,按此浓度si-r-HIF-1α和脂质体转染的心肌细胞最佳转染标本.结论 HIF-1α siRNA可显著降低成年大鼠心肌细胞中HIF-1α表达,而且转染效率很高.
목적 관찰RNA간우(RNAi)침묵성년대서심기세포결양유도인자-1α(HIF-1α)적실험가능성화전염효솔.방법 구건파향HIF-1α소간우RNA기인,병전염도성년대서세포중.반전록-취합매련반응(RT-PCR)검측성년대서심기세포적HIF-1 αmRNA수평변화,새서람(MTT)법검측불동농도적작용하대서심기세포활력,전염후24h내형광현미경검측전염대조적전염효과.결과 전염소간우RNA(siRNA)결과현시si-r-HIF-1α적침묵유효성체도73％,안차농도si-r-HIF-1α화지질체전염적심기세포최가전염표본.결론 HIF-1α siRNA가현저강저성년대서심기세포중HIF-1α표체,이차전염효솔흔고.
Objective To observe interference RNAi silencing of adult rat cardiomyocytes hypoxia-inducible factor-1 alpha (HIF-1 α) of the experimental possibilities and transfection efficiency.Methods Construction of small interfering RNA (siRNA) targeting HIF-1 α genes and transfected into cells of adult rats.Reverse transcriptase polymerase chain reaction (RT-PCR) testing HIF-1α mRNA levels in adult rat cardiac myocytes,methyl thiazol tetrazolium (MTT) method for detection of myocardial cells of rats in different concentrations of cells,transfected with fluorescence microscopy to detect within 24 hours after transfection effect of transfection with control.Results siRNA transfection results showed effectiveness about 73％ si-r-HIF-1α silence,at which concentration si-r-HIF-1α and liposome transfection of myocardial cells transfected with best samples.Conclusion HIF-1 α siRNA can significantly reduce the HIF-1 alpha expression in adult rat cardiomyocytes,and very high transfection efficiency.