孕妇循环外泌体miR-122表达在子痫前期诊断中的临床价值

The Diagnosis Value of Circulating Exosomal MiR-122 in Pregnant Women for Preeclampsia

目的研究母体循环血清外泌体miR-122表达对子痫前期的诊断价值。方法选取2018年3月至2020年4月在我院妇产科确诊的369例子痫前期孕妇作为研究对象,包括早发型(20~34孕周)子痫前期(EOPE)249例作为EOPE组,晚发型(>37孕周)子痫前期(LOPE)120例作为LOPE组。确诊时采集血样提取外泌体,另外分娩后采集胎膜组织。同时选取50例正常孕妇作为正常对照组,分别于20~34孕周和34孕周~分娩前采集静脉血样本以及分娩时的胎膜组织标本用作对照分析。采用实时荧光定量PCR法检测血清外泌体中和胎膜组织中miR-122表达,并采用甲基化特异性PCR检测胎膜组织miR-122甲基化状态。结果与正常对照组比较,EOPE组和LOPE组孕妇胎膜组织或血清外泌体中miR-122表达量升高,而组织miR-122基因甲基化率则降低(P<0.05)。另外组织miR-122甲基化亚组胎膜组织和血清外泌体miR-122表达量均低于非甲基化亚组(P<0.05)。重度亚组孕妇胎膜组织和血清外泌体miR-122表达量均高于轻度亚组(P<0.05)。经非条件Logistic回归分析,血清外泌体miR-122表达升高是EOPE和LOPE发生的独立危险因素(P<0.05)。经ROC曲线分析,血清外泌体miR-122用于EOPE和LOPE诊断的曲线下面积(AUC)分别为0.918(95%CI:0.812~0.990)、0.868(95%CI:0.776~0.973)。同样用于诊断EOPE组和LOPE组重度子痫前组患者的AUC分别为0.923(95%CI:0.817~0.991)、0.782(95%CI:0.639~0.870)。结论EOPE和LOPE孕妇血清外泌体中miR-122表达量普遍增加,这与胎膜组织中miR-122去甲基化有关。血清外泌体miR-122水平有望成为子痫前期诊断和评估疾病严重程度的重要生物标志物。

Objective To investigate the diagnosis value of circulating exosomal miR-122 in pregnant women for preeclampsia.Methods A total of 369 preeclampsia pregnant women from January,to August,2019 were enrolled for the study,including 249 cases as the early onset-preeclampsia(EOPE)group(20-34 weeks)and 120 cases as the late onset-preeclampsia(LOPE)group(>37 weeks).Blood samples were taken at the time of diagnosis and exosomes were collected.Fetal membranes were also collected after delivery.During the same period,50 normal pregnant women were selected as the control group,and venous blood samples were collected during 20~34 gestational weeks and after 34 gestational weeks to parturition,and fetal membrane tissue samples were collected at parturition.The expression of miR-122 in serum exosomes and fetal membranes was detected by real-time fluorescent quantitative PCR,and the methylation status of miR-122 in fetal membranes was detected by methylation-specific PCR.Results Compared with the normal control group,the expression of miR-122 was increased in the EOPE group and LOPE group,but the methylation rate was decreased(P<0.05).And the expression of miR-122 in fetal membranes and serum exosomes in the miR-122 methylated group was lower than that in the non-methylated group(P<0.05).The expression of miR-122 in fetal membranes and serum exosomes in the severe group was higher than that in the mild group,while the methylation rate of miR-122 in the severe group was lower than that in the mild group(P<0.05).The expression of serum exosomal miR-122 was an independent risk factor for the occurrence of EOPE and LOPE(P<0.05)by non-conditional logistic regression analysis.ROC curve analysis showed that area under the curve(AUC)of serum exosomal miR-122 for the diagnosis of EOPE and LOPE were 0.918(95%CI:0.812-0.990)and 0.868(95%CI:0.776-0.973),while AUC for the diagnosis of severe EOPE or severe LOPE were 0.923(95%CI:0.817-0.991)and 0.782(95%CI:0.639-0.870),respectively.Conclusion The expressions of serum exosomal miR-122 in EOPE and LOPE pregnant women significantly increase,which are associated with miR-122 demethylation in fetal membranes.The detection of serum exosomal miR-122 level is expected to be an important biomarker for the diagnosis and assessment of the severity of preeclampsia.