应用siRNA抑制由醛固酮诱导的大鼠肥大心肌细胞中钙调神经磷酸酶Aβ mRNA的表达(PDF)
《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]
- 期数:
- 2005年第4期
- 页码:
- 313-316
- 栏目:
- 基础研究
- 出版日期:
- 2005-09-05
文章信息/Info
- Title:
- Inhibition of the expression of calcineurin Aβ mRNA initiated by siRNA in rat hypertrophic cardiomyocytes induced by aldosterone
- Author(s):
- LIU Li; XIE Xiaohua; CHANG Lianqing
- General Hospital of PLA, Beijing 100853, China
- Keywords:
- cardiomyocyte; calcineurin; siRNA; RNAi; aldosterone
- 分类号:
- R335
- DOI:
- -
- 文献标识码:
- A
- 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（Ald）诱导的大鼠心肌细胞肥大时其Aβ mRNA表达水平。方法 在Ald诱导心肌细胞肥大模型上同时转染CaN Aβ mRNA靶标siRNA及其阴性对照进行转染。通过免疫荧光染色及RTPCR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 Ald组CaN Aβ mRNA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，RTPCR检测结果显示siRNA组 CaN Aβ mRNA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siRNA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。
- Abstract:
- AIM To investigate posttranscriptional 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 RTPCR. 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. RTPCR 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.
参考文献/References
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备注/Memo
- 备注/Memo:
- 收稿日期：2005-01-05.基金项目：“十五”军队医药卫生科研基金重点课题资助（02Z0010）
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