肥厚型心肌病患者超极化激活环核苷酸门控阳离子通道mRNA的表达

Detection of hyperpolarization-activated cyclic nucloetide-gated cation channel mRNA expression in hypertrophic cardiomyopathy

目的探讨肥厚型心肌病患者超极化激活环核苷酸门控阳离子通道(HCN4)的表达。方法构建重组质粒,用梯度稀释的质粒模板建立SYBRGreen荧光定量逆转录-聚合酶链式反应(RT-PCR)法检测HCN4的标准曲线,利用建立的RT-PCR体系对肥厚型心肌病和正常心肌组织标本的左室HCN4mRNA进行检测。结果成功构建了质粒重组子,并建立用SYBRGreen荧光染料在RNA水平检测HCN4通道的标准曲线。HCN4通道在肥厚型心肌病中的表达水平高于正常心肌。结论肥厚型心肌病患者HCN4mRNA的表达较正常人增高。

Objective To establish a fluorogenie quantitative RT-PCR （FQ-RT-PCR） method with SYBRGreen dye for detecting the expression of hyperpolarization-activated cyclic nueloetide-gated cation channel （HCN4） mRNA in hypertrophic eardiomyopathy. Methods To construct the recombinant plasmid and establish the standard curve of HCN4 by FQ-RT-PCR using a serial dilution of plasmids. The expression of HCN4 gene in the hypertrophic cardiomyopathy and nor- mal cardie muscles were detected by the established FQ-RT-PCR assay. Results The recombinant plasmid was successfully constructed and the standard curve to detect the expression of the HCN4 gene was established. HCN4 expression in the hypertrophic cardiomyopathy was higher than that in the normal cardic muscles. Conclusion The expression level of HCN4 mRNA in the hypertrophic cardiomyopathy patients was higher than control group.