miRNA表达谱在缺氧缺血性脑病新生儿血浆中的变化及意义

Expression profile of plasma mi RNA in neonates hypoxic-ischemic encephalopathy and its clinical significance

目的筛选新生儿缺氧缺血性脑病(HIE)相关的miRNA,探讨miRNA在HIE诊断、治疗和预后中的指导意义。方法选择HIE新生儿117例,根据病情分为轻度组69例和中重度组48例;选择同期分娩的健康足月新生儿30例为健康对照组。利用miRNA芯片检测3组新生儿血浆miRNA表达谱的差异,从中挑选6个miRNA进行Real-time PCR验证,比较各组血浆miRNA表达并分析与HIE诊断、治疗和预后的关系。结果 miRNA芯片检测HIE组共有55种miRNA表达有明显差异,有29种miRNA表达上调,26种表达下调。与健康对照组比较,HIE组血浆中miR-4472、miR-5195-3p、miR-6794-5p表达明显上调,let-7b-5p表达明显下调,差异均有统计学意义(均P<0.01)。HIE中重度组miR-4472、miR-5195-3p、miR-6794-5p表达显著高于轻度组,而let-7b-5p表达显著低于轻度组,差异均有统计学意义(均P<0.01)。与治疗前比较,治疗后HIE轻度组和中重度组miR-4472、miR-5195-3p、miR-6794-5p表达量下降,let-7b-5p表达量上升,差异均有统计学意义(均P<0.01)。ROC曲线分析显示miR-4472、miR-5195-3p、miR-6794-5p和let-7b-5p 4种miRNA的AUC分别为0.958、0.938、0.986和0.969。miR-4472、miR-6794-5p高表达组后遗症发生率明显高于低表达组(均P<0.05),而miR-5195-3p、let-7b-5p与HIE预后无相关性(均P>0.05)。结论 HIE患儿血浆中miR-4472、miR-5195-3p、miR-6794-5p表达水平显著升高,而let-7b-5p表达水平显著下降,其中miR-4472、miR-6794-5p可能是预测HIE预后的生物标志物。

Objective To screen the relevant plasma miRNA in neonates hypoxic-ischemic encephalopathy（HIE）, and to investigate its clinical significance. Methods A total of 117 neonates with HIE（including 69 mild cases and 48 moderate or severe cases） were enrolled in the study, 30 full term healthy neonates served as healthy controls. The expression profile of plasma miRNA in three groups was examined with miRNA microarray, then 6 miRNAs were chosen for further validation by Real-time PCR. The expression of miRNA was compared among these groups, and the relationship between miRNA expression and diagnosis, treatment and prognosis of HIE was analyzed. Results The miRNA microarray revealed that 55 miRNA were differentially expressed in plasma of HIE group, 29 miRNA were up-regulated and 26 miRNA were down-regulated. Compared to healthy controls, the expression of mi R-4472, mi R-5195-3p, mi R-6794-5p was increased significantly in HIE group, while the expression of let-7b-5p decreased significantly（all P〈0.01）. The expression of mi R-4472, mi R-5195-3p and mi R-6794-5p in moderate or severe HIE group was significantly higher, and the expression of let-7b-5p was lower than those in mild HIE group（all P〈0.01）. In mild HIE group and moderate or severe HIE group, the expression mi R-4472, mi R-5195-3p and mi R-6794-5p was decreased, and the expression of let-7b-5p was increased markedly after successful treatment（all P〈0.01）. ROC curve showed that the area under the curve（AUC） of these 4 miRNAs were 0.958, 0.938, 0.986 and 0.969, respectively. The incidence of sequelae in HIE neonates with highly expressed mi R-4472 and mi R-6794-5p was significantly higher than that with low expression（both P〈0.05）. The expression of mi R-5195-3p and let-7b-5p was not correlated with prognosis of HIE neonates（both P〈0.05）. Conclusion The expression of mi R-4472, mi R-5195-3p, mi R-6794-5p in HIE group are significantly up-regulated, while let-7b-5p is down-regulated. The mi R-4472 and mi R-6794-5p might be used as biomarkers for prognosis of HIE.