成人暴发性心肌炎中循环微小RNA-29b的表达及其意义

目的 探讨成人暴发性心肌炎(FM)循环微小RNA(miRNA)的表达谱特征及其诊断的潜在标志物。方法 通过微阵列分析检测循环miRNA表达谱,实时荧光定量PCR方法验证miRNA表达水平,通过KEGG通路分析循环miRNA在FM中的关键作用,对FM患者的miRNA与心功能参数进行相关分析,绘制受试者工作特征(ROC)曲线分析循环miRNA表达在FM诊断中的灵敏度和特异度。结果 与健康对照组相比,FM患者组血浆中miR-29b(t=18.925,P<0.001)和miR-125b(t=5.981,P=0.029)的表达水平显著上调。与治疗前相比,治疗后FM患者miR-29b(t=12.943,P<0.001)和miR-125b(t=14.016,P<0.001)的表达水平显著下调。KEGG通路富集分析显示,miR-29b靶点参与炎症反应和细胞凋亡途径。细胞增殖与凋亡检测显示,与miR-125b模拟物相比,转染miR-29b模拟物对心肌细胞凋亡的诱导作用更加明显(χ²=6.168,P=0.047),而两组细胞增殖抑制情况差异无统计学意义(χ²=1.452,P=0.417)。线性相关分析显示,miR-29b和miR-125b表达水平与左心室射血分数呈负相关(r=-0.67,P=0.071;r=-0.49,P=0.003),与心肌肌钙蛋白Ⅰ浓度(r=0.61,P=0.019;r=0.52,P=0.016)、干扰素β(IFN-β)浓度(r=0.42,P=0.014;r=0.36,P=0.021)以及心肌水肿面积(r=0.86,P=0.005;r=0.73,P=0.013)呈显著正相关。ROC曲线分析显示,与miR-125b相比,miR-29b诊断FM具有较高的灵敏度(93.6%比89.2%;t=0.896,P=0.795)和特异度(72.4%比59.6%;t=9.478,P=0.002)。结论 循环miR-29b可作为FM诊断的潜在生物标志物。

Expression and Significance of Circulating microRNA-29b in Adult Fulminant Myocarditis

Objective To explore the expression profiles of circulating microRNA(miRNA)and potential markers for the diagnosis of adult fulminant myocarditis(FM). Methods The expression profiles of circulating miRNA were determined by microarray analysis and verified by real-time quantitative PCR.The key role of circulating miRNA in FM was determined via KEGG pathway enrichment.The correlations between miRNA and cardiac function parameters in patients with FM were analyzed.The receiver operating characteristic(ROC)curve was established to evaluate the sensitivity and specificity of circulating miRNA in the diagnosis of FM. Results Compared with healthy controls,the FM patients had up-regulated expression levels of miR-29b(t=18.925,P<0.001)and miR-125b(t=5.981,P=0.029)in the plasma.After treatment,the expression levels of miR-29b(t=12.943,P<0.001)and miR-125b(t=14.016,P<0.001)were significantly down-regulated.KEGG pathway enrichment showed that the targets of miR-29b were involved in inflammatory response and apoptosis pathways.The results of cell proliferation and apoptosis assay demonstrated the transfection of miR-29b mimic had a more significant inducing effect on cardiomyocyte apoptosis than that of miR-125b mimic(χ ²=6.168,P=0.047),whereas there was no significant difference in the inhibition of cell proliferation between the two groups(χ²=1.452,P=0.417).The expression levels of miR-29b and miR-125b were negatively correlated with left ventricular ejection fraction(r=-0.67,P=0.071;r=-0.49,P=0.003).They were positively correlated with cardiac troponin I level(r=0.61,P=0.019;r=0.52,P=0.016),interferon β level(r=0.42,P=0.014;r=0.36,P=0.021),and myocardial edema area(r=0.86,P=0.005;r=0.73,P=0.013).The ROC curve analysis demonstrated that miR-29b had higher sensitivity for the diagnosis of FM(93.6% vs.89.2%;t=0.896,P=0.795)and specificity(72.4% vs.59.6%;t=9.478,P=0.002)than miR-125b. Conclusion The circulating miR-29b may be a potential biomarker for the diagnosis of FM.