质粒shRNA体内干扰Klotho基因对窦房结通道基因的影响

Study on the Effect of Klotho Gene Interferred by Plasmid-mediated Short Hairpin RNA(shRNA) on Sinoatrial Node Pacing Channel Gene

通过质粒shRNA体内干扰,研究Klotho基因与窦房结起搏通道相关基因HCN4及HCN2之间的关系,为病窦综合征的研究提供新思路。取C57BL/6J小鼠20只,分为4组,每组5只,分别为:质粒shRNA 24h组、质粒shRNA 12h组、生理盐水24h组、生理盐水12h组。质粒shRNA组经尾静脉注射质粒shRNA 50μL(1μg质粒/μL),生理盐水组经尾静脉注射生理盐水50μL。分别于注射12h及24h后取窦房结周围组织,行RT-PCR检测各组小鼠的Klotho、HCN2、HCN4基因的mRNA水平。RT-PCR结果显示:与生理盐水12h组比较,shRNA 12h组的klotho、HCN4和HCN2的mRNA表达量明显降低,均有统计学差异(P<0.05)。以上结果提示,小鼠Klotho基因和窦房结起搏基因可能存在一定关系。

The study was aimed to assess the effect of Klotho gene and sinoatrial node pacing channel gene （HCN4 and HCN2） for studying sick sinus syndrome, with Klotho gene under the interference of Plasmid-mediated short hairpin RNA. Twenty-five C57BL/6J mice were divided into four groups, i. e, plasmid shRNA 24h group, plasmid shRNA 12h group, sodium chloride 24h group and sodium chloride 12h group. Plasmid shRNA 50vL （lvg/t^L） and sodium chloride 50~1 were respectively injected according to mice vena caudalis into those in plasmid shRNA group and sodium chloride group. After 12h or 24h respectively, all mice were executed and their sinoatrial node tissues were cut. The mRNA of Klotho, HCN4 and HCN2 gene were detected by RT-PCR. The results of RT-PCR showed that Klotho, HCN4 and HCN2 mRNA levels were lower compared with those in sodium chloride 12h group after i2h interference interval. The results indicated that there might be the a certain relationship between Klotho gene and sinoatrial node pacing channel gene.