摘要
目的:本研究通过应用RNA干扰的方法,观察了钙调神经磷酸酶(CaN)在醛固酮(A ld)诱导的大鼠心肌细胞肥大时其AβmRNA表达水平。方法:在A ld诱导心肌细胞肥大模型上同时转染CaN AβmRNA靶标siRNA及其阴性对照进行转染。通过免疫荧光染色及RT-PCR方法检测CaN Aβ蛋白及mRNA的表达,同时测定心肌细胞面积变化。结果:A ld组CaN AβmRNA表达较对照组显著升高,细胞表面积显著增加;CaN AβmRNA靶标siRNA组细胞浆中绿色荧光表达比siRNA阴性对照组少,即CaN蛋白表达量siRNA组较siRNA阴性对照组减少,RT-PCR检测结果显示siRNA组CaN AβmRNA表达较siRNA阴性对照组受到明显抑制,细胞面积siRNA阴性对照组较siR-NA组增加。结论:siRNA可抑制钙调神经磷酸酶AβmRNA的表达,对于心肌细胞中基因功能的研究,RNA干涉可以作为应用工具。
AIM： To investigate post-transcriptional gene silencing of calcineurin Aβ mRNA initiated by siRNA in neonatal rat cardiomyocyte induced by aldosterone （ Ald）. METHODS ：Ald was used to stimulate neonatal rat cardiomyocytes, at the same time, calcineurin Aβ mRNA was transfected to target siRNA and its negative control. The expression of calcineurin Aβ mRNA was detected by immunofluorescence and RT-PCR. RESULTS ： The expression of calcineurin Aβ mRNA and the area of the cells of the Ald group were more and larger than those of the control group. The expression of immunofluorescence of the siRNA target group was less than that of the siRNa control group. RT-PCR examination showed that the expression of CaN Aβ mRNA of the siRNA group was greatly inhibited, and the area of the cells was smaller, as compared with the siRNA negative control. CONCLUSION： Calcineurin plays a role in the progression of cardiac hypertrophy induced by Ald. The calcineurin Aβ mRNA is markedly inhibited by RNAi. So RNA can be a very effective method by which to study the gene functions of cardiomyocytes.
出处
《心脏杂志》
CAS
2005年第4期313-316,共4页
Chinese Heart Journal
基金
"十五"军队医药卫生科研基金重点课题资助(02Z0010)
