急性心肌梗死和急性脑梗死miRNA疾病标志物的初步筛查

Preliminary Screening of mi RNA Disease Markers for Acute Myocardial and Cerebral Infarctions

目的确定一种可以作为急性心肌梗死(AMI)或急性脑梗死(ACI)疾病诊断标志物的新的mi RNA组合。方法根据文献与公开的互联网数据库,选择高频出现的mi RNA做为候选基因,收集AMI患者外周血样本13例、ACI患者外周血样本11例和健康志愿者样本20例。利用试剂盒提取这些样本中的游离RNA,进行反转录及实时定量PCR测定,检测所有样本中候选mi RNA的表达情况。对2个疾病组和健康对照组中候选mi RNA的表达差异进行统计学分析。结果经过筛查,AMI组中出现频率最高的mi RNA有mi RNA1、mi RNA499a和mi RNA133a,ACI组中出现频率最高的mi RNA有let7i、mi RNA16和mi RNA223。2个疾病组中6种mi RNA的表达量均显著高于健康对照组(P<0.01)。AMI组mi RNA1和mi RNA499a的表达量分别为ACI组的4.82倍和3.50倍(P<0.01),而mi RNA133a的表达量无统计学差异(P>0.05)。ACI组与AMI组比较,mi RNA16和mi RNA233的表达量无统计学差异(P>0.05),ACI组let7i的表达量为AMI组的2.61倍(P<0.05)。结论人外周血游离RNA中6种mi RNA均异常升高可以作为AMI或ACI的初步诊断标志物,在此基础上,mi RNA1和mi RNA499a的表达量达到11~20倍以上时,对AMI的发病具有辅助诊断及预警作用;let7i的表达量达到6倍以上时,对ACI的发病具有辅助诊断及预警作用。

Objective To identify a new miRNA combination that could be used as a diagnostic marker for acute myocardial infarction （AMI） or acute cerebral infarction （ACI）. Methods We surveyed the literatures and databases and selected miRNAs with a high frequency in AMI or ACI as candidate genes. We collected peripheral blood samples from 13 patients with AMI, 11 patients with ACI, and 20 healthy volunteers. RNA was extracted from these samples, and gene expression levels were determined by reverse transcription and real-time quantitative PCR. The differences in the expression of the candidate miRNAs were then analyzed. Results The frequencies of miRNA1, miRNA499a, and miRNA133a were high- er in the AMI group, whereas the frequencies of let7i, miRNA16, and miRNA223 were higher in the ACI group. These six miRNAs were signifi- cantly higher in the two disease groups than in the control group （P 〈 0.01 ）. The levels of miRNA1 and miRNA499a were 4.82- and 3.50-times higher in the AMI group, respectively （beth P 〈 0.01 ）, when compared with the ACI group. The levels of let7i in the ACI group were 2.61-times higher than that in the AMI group （P 〈 0.05 ）. There were no significant differences in the expression levels of miRNA 133a, miRNA 16, and miR- NA233 between the AMI and ACI groups （P 〉 0.05 ）. Conclusion The abnormal increase in six miRNAs in human peripheral blood could be used as a marker for the diagnosis of AMI or ACI. The expression levels of miRNA 1 and miRNA499a were 11- to 20-times higher than control lev- els, and the expression levels of let7i were 6-times higher than control levels, which could be used to predict the risk of AMI and ACI, respectively.