miRNA表达谱在产前诊断胎儿先天性心脏病中的研究  

The Study of MiRNA Expression Profile in Prenatal Diagnosis of Fetal Congenital Heart Disease

作　　者：杨微微[1] 任晨春[1] 常颖[2] 王文靖[1] 鞠明艳 姚立英[2] 赵晓敏[2] 赵丹阳 YANG Wei-wei;REN Chen-chun;CHANG Ying;WANG Wen-jing;JU Ming-yan;YAO Li-ying;ZHAO Xiao-min;ZHAO Dan-yang(Department of Laboratory,Tianjin Central Hospital of Gynecology Obstetrics,Tianjin 300100,China;Prenatal Diagnosis Center,Tianjin Central Hospital of Gynecology Obstetrics,Tianjin 300100,China;Tianjin Union Medical Center,Tianjin 300121,China)

机构地区：[1]天津市中心妇产科医院检验科,300100 [2]天津市中心妇产科医院产前诊断中心,300100 [3]天津市人民医院

出　　处：《国际妇产科学杂志》2024年第3期342-346,共5页Journal of International Obstetrics and Gynecology

基　　金：天津市卫生健康委员会科技项目(ZC20117);天津市科技计划项目(21JCQNJC01860);天津市医学重点学科(专科)建设项目(妇产科学TJYXZDXK-043A)。

摘　　要：目的:探究微小RNA(microRNA,miRNA)表达谱在产前诊断胎儿先天性心脏病(congenital heart disease,CHD)中的应用。方法:收集2021年1月—2022年12月于天津市中心妇产科医院就诊的30例超声确诊为CHD的孕妇(病例组)和同期10例要求行羊水穿刺的孕妇(对照组),用Illumina测序平台对2组孕妇的羊水上清进行全转录组测序,2组孕妇的全部miRNA进行归一化,分析差异表达的miRNA。从差异表达的miRNA中挑选P<0.05和|log2 FC|>3(差异倍数,Fold Change,FC)的miRNA再在羊水和外周血中进行实时荧光定量聚合酶链反应(real time fluorescence quantitative polymerase chain reaction,RT-qPCR)验证,比较羊水中miRNA测序与RT-qPCR的差异倍数,挑选外周血与羊水表达调控方向一致的miRNA。结果:共发现138个差异表达miRNA,其中85个上调,53个下调。进一步挑选出了15个差异表达的miRNA,羊水中miRNA测序与RT-qPCR结果比较相一致。外周血与羊水中表达调控方向一致的miRNA有2个,分别为miR-222-3p和miR-189-5p,这2个miRNA在病例组母血中表达量较对照组显著上升(均P<0.05)。结论:母血中miRNA作为新的血清学标志物可以初步应用于筛查胎儿CHD。Objectives:To explore the application of microRNA(miRNA)expression profile in prenatal diagnosis of fetal congenital heart disease(CHD).Methods:30 pregnant women diagnosed with CHD by ultrasound(case group)and 10 pregnant women who required amniocentesis(control group)were collected from January 2021 to December 2022 at Tianjin Central Hospital of Gynecology Obstetrics.The amniotic fluid supernatant of the two groups of pregnant women was sequenced by Illumina sequencing platform.All miRNAs of the two groups of pregnant women were normalized and the differentially expressed miRNAs were analyzed.The miRNA with P<0.05 and|log2 FC|>3(Fold Change,FC)were selected from the differentially expressed miRNAs for further validation by real time fluorescence quantitative polymerase chain reaction(RT-qPCR)in amniotic fluid and peripheral blood.The difference between miRNA sequencing and RT-qPCR in amniotic fluid was compared,and the miRNAs with consistent expression regulation direction in peripheral blood and amniotic fluid were selected.Results:A total of 138 differentially expressed miRNAs were found,of which 85 were up-regulated and 53 were down-regulated.Further,15 differentially expressed miRNAs were selected,and the results of miRNA sequencing in amniotic fluid were consistent with those of RT-qPCR.There were two miRNAs with the same expression and regulation direction in peripheral blood and amniotic fluid,which were miR-222-3p and miR-189-5p,respectively.The expression of these two miRNAs in maternal blood of the case group was significantly higher than that of the control group,and the difference was statistically significant(all P<0.05).Conclusions:As a new serological marker,miRNA in maternal blood can be preliminarily applied to the screening of fetal CHD.

关 键 词：心脏缺损 先天性 微RNAS 产前诊断 羊膜腔穿刺术 非侵入性产前检测 实时聚合酶链反应 

分 类 号：R714.5[医药卫生—妇产科学]