不同LOX-1基因型急性心肌梗死患者血清6种微小RNAs的表达特点

Expression of 6 serum microRNAs in patients with acute myocardial infarction and different LOX-1 genotype

目的探讨急性心肌梗死患者的6个微小RNA(microRNA,miRNA):miRNA-1、miRNA-21、miRNA-133、miRNA-199、miRNA-208、miRNA-499的表达水平及其在不同凝集素样氧化型低密度脂蛋白受体-1(lectin-likeoxidized low-density lipoprotein receptor-1,LOX-1)基因型患者的表达特点。方法采用新型分子信标探针和实时荧光定量聚合酶链反应(polymerase chain reaction,PCR)技术检测120例急性心肌梗死患者和80名正常健康人血清中以上6个miRNAs的表达水平;采用实时荧光定量PCR技术检测LOX-1基因,并进行各基因型以上miRNAs表达水平的比较;利用多指标同步分析(flexible multi-analyte profiling,xMAP)液态芯片技术验证以上6个miRNAs的表达水平。结果 LOX-1 GG基因型的血清miRNA-208在急性心肌梗死发生后50 min表达升高,miRNA-21、miRNA-133、和miRNA-199在急性心肌梗死发生后2 h表达升高,血清miRNA-499在急性心肌梗死发生后4 h表达升高,各自在不同时间下降,低水平表达维持到急性心肌梗死发生后69 h。携带LOX-1N基因型的急性心肌梗死患者的miRNA-21、miRNA-133、miRNA-199、miRNA-208、miRNA-499表达下调,miRNA-1表达上调。新型分子信标实时荧光定量PCR技术和xMAP液态芯片技术检测的miRNA-21(r=0.810,P<0.05),miRNA-208(r=0.717,P<0.05),miRNA-133(r=0.631,P<0.05),miRNA-499(r=0.604,P<0.05)表达呈正相关。结论miRNA-21、miRNA-133、miRNA-199、miRNA-208、miRNA-499在急性心肌梗死患者的异常表达可作为早期诊断心肌梗死的生物标志物,为早期诊断心肌梗死提供新的检测手段。

Objectives To investigate the expression and characteristics of 6 microRNAs （miRNAs） （miRNA-1 ,miRNA-21, miRNA-133,miRNA-199,miRNA-208,miRNA-499） in patients with acute myocardial infarction （AMI） and different oxidized low-density lipoprotein receptor-1 （LOX-1 ） genotype. Methods Serum levels of the 6 miRNAs in 120 patients with AMI and 80 normal people were determined by molecular beacon （MB） and fluorescence real-time polymerase chain reaction （PCR）. LOX-1 of the 120 patients with AMI was detected by fluorescence real-time PCR. Serum levels of the 6 miRNAs in patients with AMI and different LOX-1 genotype were compared. Expression of the 6 miRNAs was identified by flexible multi-analyte profiling （xMAP）. Results Serum level of miRNA-208 of LOX-1 GG genotype significantly up-regulated 50 min after AMI. Serum levels of miRNA-21, miRNA-133 and miRNA-199 significantly up-regulated 2 h after AMI. Serum levels of miRNA-499 up-regulated 4 h after AMI. They significantly down-regulated in different time and maintained at low level until 69 h after AMI. Serum levels of miRNA-21, miRNA-133, miRNA-199, miRNA-208 and miRNA-499 significantly down-regulated in patients with AMI with LOX-1N gene, and miRNA-1 significantly up-regulated. MiRNA-21 （r=0.810,P〈0.05）, miRNA-208 （r=0.717,P〈0.05）, miRNA-133 （r=0.631, P〈0.05 ） and miRNA-499 （r=0.604, P〈0.05） tested by xMAP positively correlated with those tested by MB and fluorescence real-time PCR. Conclusions MiRNA-21, miRNA-133, miRNA-199, miRNA-208 and miRNA-499 are potential diagnostic serum marker of AMI, which provides the basis for new detection means of early diagnosis of AMI.