慢性心房颤动患者心房肌mink亚单位基因mRNA水平的定量检测

目的：心房颤动是目前最为常见的快速性心律失常之一。mink是心肌细胞延迟整流钾通道（Iks）的重要辅助亚单位,在Iks发挥正常功能中起着重要的作用,同时参与心肌细胞其它离子通道电流的调控。本研究采用实时荧光定量PCR技术通过对房颤时mink基因mRNA水平进行定量研究,探讨mink在房颤中的作用和潜在调控机制。方法：①16例风湿性心脏病（rheumaticheartdisease,RHD）患者分为2组：窦性心律组（SR组）8例、房颤心律组（AF组）8例,取其常规切除的右心耳组织作为研究对象;②提取心房肌组织总RNA,通过RT-PCR和TA克隆技术构建含有目的基因片段的质粒标准品;③以GAPDH为内参照基因,采用SYBRGreenⅠ法实时荧光定量PCR技术检测慢性心房颤动和窦性心律患者mink基因mRNA水平。结果：①标准曲线和熔解曲线：标准曲线线性关系良好,相关系数R2〉0.99。熔解曲线呈明显单峰状,退火温度与预测值基本一致;②SR组和AF组mink/GAPDH比值分别为0.1890±0.0721（n=8）和0.1413±0.0954（n=8）,AF组mink基因mRNA水平低于SR组,P〈0.05。结论：与窦性心律患者相比,心房颤动患者mink基因明显下调,mink基因表达量的改变可能参与房颤重构的分子基础。

REAL-TIME FLUORESCENT QUANTITATION PCR DETECTS MINK GENE MRNA EXPRESSION LEVEL IN ATRIAL MYOCYTES OF HUMAN CHRONIC ATRIAL FIBRILLATION

Objective： Atrial Fibrillation （AF） is one of the most common tachyar rhythmia. This study is to detect the mink mRNA level in AF by real-time PCR, and discuss the role of mink in AF. Methods： ①30 right atrial appendage samples from patients with rheumatic heart disease （RHD） were divided into two groups： the AF group （n=8） and the sinus rhythm（SR） group （n=8）. ②The total RNA was acquired from the atrial tissues, and the standard preparations were reconstructed from plasmids including the target gene by RT-PCR and TA cloning. ③ The mink gene mRNA level was compared between the two groups by SYBR Green I real-time PCR with the GAPDH as the house keeping gene. Results： ①the standard curve and the melting curve： the standard curves had good linear correlation, the contiguous coefficient R20.99. The shape of the melting curves was unimodality. The Tm values of the products were very approximate to the predictive values. ②The respective ratio of KChIP2/ GAPDH between the two groups was 0.1890±0.0721 （n=8） and 0.1413±0.0954 （n=8）, P0.05; Conclusion：Compared to the RHD patients with SR, the mink gene mRNA level in the patients with chronic AF was down-regulated significantly. The down-regulation of the mink gene is one of the molecular bases on AF.