肝素对血浆和组织microRNA的PCR扩增效率的影响  

Impact of Intravenous Heparin on the Amplification Efficiency of PCR on microRNAs in Plasma and Tissues

作　　者：陈心培 张瑜[1] 马永江[2] 吴丹[1] 陈桂颖[3] 孟鹤[3] 单兆亮[1] 

机构地区：[1]解放军总医院心内科,北京100853 [2]解放军304医院心内科,北京100037 [3]天津市胸科医院胸内科,天津300151

出　　处：《现代生物医学进展》2014年第21期4041-4043,4082,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金面上项目(81270308);北京市自然科学基金项目(7122173)

摘　　要：目的:研究肝素对血浆和组织microRNA(miRNA)提取及PCR扩增效率的影响。方法:将杂种犬9只,雌雄不分,随机分为两组,对照组(n=5)和实验组(n=4),实验组静脉注射肝素十分钟后取静脉血(用不含肝素的真空采血管采集)及左心室组织,对照组注射相对应量的生理盐水后步骤同实验组,应用miRNA试剂盒分别提取实验组和对照组血浆和组织的miRNA,逆转录成cDNA后进行实时定量PCR分析。结果:实验组与对照组比较血浆miRNA Ct值明显增高,其差异具有统计学意义(P<0.01),组织miRNA Ct值无明显变化。结论:静脉注射肝素短时间内对血浆miRNA的PCR扩增效率造成明显影响,对组织miRNA提取及PCR扩增效率无明显影响。Objective： To study the impact of intravenous heparin on the amplification efficiency of PCR on microRNAs in plasma and tissues. Methods： Nine dogs were randomly divided into control group （n=5） and intravenous heparin group （n=4）. We collected venous blood, tissues of left ventricle 10 mins after heparin administration in intravenous heparin group, so as the control group except injected heparin. The extraction of miRNA was performed by specific miRNA kit. miRNA was converted into DNA for RT-PCR analysis. Results： Compared with control group, the intravenous heparin group Ct values of miRNA in plasma increased significantly （P 〈0.01. There was no statistical significance in tissues. Conclusion： Heparin has a significant influence on the amplification efficiency of PCR on microRNAs in plasma and has no obvious effect on that in tissues.

关 键 词：肝素 MICRORNA PCR 

分 类 号：Q95-3[生物学—动物学] Q503