miR-130a和miR-125b在川崎病外周血细胞中的表达及意义

Plasma miR-130a and miR-125b as biomarkers of coronary artery dilatation in Kawasaki disease

目的通过检测miR-130a和miR-125b在伴或不伴冠脉扩张(CAD)川崎病(KD)患儿与正常健康儿童外周血单核细胞(PBMC)中的表达差异,探讨两者可否作为KD和CAD诊断及预后评估的全新血清生物标志物。方法 利用基因芯片技术筛选出KD患儿与正常健康儿童之前具有差异表达的miRNAs,通过生物信息学分析,确定候选基因。进一步选取KD患儿30例(伴或不伴CAD各15例)、正常健康儿童15例,采用茎环RT-PCR的方法 ,验证候选基因miR-130a和miR-125b在PBMC中的表达情况。结果 (1)通过基因芯片技术,分析KD患儿与正常健康儿童PBMC中存在明显差异表达的miRNA共63条,经过生物信息学分析,确定候选基因;(2)通过茎环RT-PCR验证发现miR-130a和miR-125b在患儿IVIG治疗前表达明显下调,而IVIG治疗后表达明显上调(P<0.05);(3)伴CAD的KD患儿miR-130a及miR-125b表达较不伴CAD明显升高,且呈正相关性(R^2=0.734,mir-130a;R^2=0.709,miR-125b);(4)ROC曲线分析表明miR-130a(特异度87.5%和敏感度77.5%)和miR-125b(特异度83.3%和敏感度76.6%)对KD及CAD的判断有较高的特异度和敏感度。结论 PBMC中的miR-130a和miR-125b参与了KD的发生与发展,可作为KD及CAD诊断及预后评估的一项全新血清生物标志物,并为KD冠脉扩张的防治提供新的思路。

Objective To investigate the expression of microRNAs （miRNAs） in peripheral blood mononuclear cel s （PBMCs） and plasma of patients with Kawasaki disease （KD）. Methods Fifteen KD pediatric patients with coronary artery di-latation （CAD）, 15 KD patients without CAD and 15 healthy children were enrolled in the study. The expression of KD- associated miRNAs in PBMC was screened by microarray. Results The results showed that 63 miRNAs were differential y expressed in PBMCs between the KD group and control group. Two miRNAs miR- 130a and miR- 125b down- regulated more than 3- fold were selected for study. The expression of miR- 130a and miR- 125b in plasma samples of KD patients was detected by stem- loop RT- PCR before and after treatment with intravenous immunoglobulin. The expression of miR- 130a and miR- 125b were higher after treatment than that before treatment （P〈0.05）. miR- 130a and miR- 125b in KD patients were correlated with coronary artery dilatation（CAD） （R2=0.734 and R2=0.709, respectively）. Receiver operating characteristic analysis indicated that miR- 130a had relatively high sensitivity and specificity to KD with CAD （87.5% and 77.5%, respectively）. For miR- 125b, these values were 83.3% and 76.6%. Conclusion miR- 130a and miR- 125b may be associated with the progression of KD and they might be used as potential biomarkers for the diagnosis and prognosis of KD patients with CAD.