Li F, Lin H, Li Y, Zhu W, Sun Y, Huang Y, Qiu Y, Qin X, Chang Q
Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2024 Nov 20;44(11):2256-2264. doi: 10.12122/j.issn.1673-4254.2024.11.24.
To investigate the role of miRNAs in maternal amniotic fluid exosomes in development of isolated ventriculomegaly (VM) in fetuses.
Amniotic fluid samples were collected from 9 cases of moderate isolated VM and 8 normal control cases to extract exosomal miRNA, and miRNA sequencing technique was used to identify differentially expressed miRNAs between the two groups. Three miRNAs with significant differential expression between the two groups, whose high expression was associated with VM, were selected for verification with RT-qPCR. Dual luciferase reporter assays were used to verify the regulatory effect of miR-122-5p on its predicted target genes and . Gene ontology (GO) and KEGG pathway analyses were performed to explore the possible roles of the top 40 significant differential miRNAs in the pathophysiology of VM.
We identified a total of 272 differentially expressed miRNAs in VM cases, including 43 up-regulated and 229 down-regulated miRNAs. The target genes of these differential miRNAs were associated with DNA and transcription factor binding, transmembrane transporter and nucleic acid binding transcription factor activity, and cell developmental process. These miRNAs were mostly enriched in the MAPK, cGMP-PKG and Wnt signaling pathways. Verification with RT-qPCR showed that miR-122-5p expression level was significantly lower in VM group than in the control group ( < 0.05), which was consistent with miRNA sequencing results; let-7b-5p expression level was significantly lower in VM group, which was contrary to miRNA sequencing result. Dual luciferase reporter assays showed that miR-122-5p was not capable of regulating or expressions.
The highly abundant differentially expressed miRNAs in maternal amniotic fluid exosomes play important roles in the occurrence of fetal VM possibly by regulating the MAPK, PI3K-Akt, Wnt and cGMP-PKG signaling pathways.
探讨微小RNA（miRNAs）在母体羊水外泌体中对胎儿孤立性脑室扩大（VM）发育的作用。
收集9例中度孤立性VM病例和8例正常对照病例的羊水样本，提取外泌体miRNA，采用miRNA测序技术鉴定两组间差异表达的miRNA。选择两组间差异表达显著且高表达与VM相关的3种miRNA，用逆转录定量聚合酶链反应（RT-qPCR）进行验证。采用双荧光素酶报告基因检测法验证miR-122-5p对其预测靶基因和的调控作用。进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路分析，以探讨前40个差异显著的miRNA在VM病理生理学中的可能作用。
我们在VM病例中总共鉴定出272个差异表达的miRNA，包括43个上调的和229个下调的miRNA。这些差异miRNA的靶基因与DNA和转录因子结合、跨膜转运体以及核酸结合转录因子活性和细胞发育过程相关。这些miRNA大多富集于丝裂原活化蛋白激酶（MAPK）、环磷酸鸟苷-蛋白激酶G（cGMP-PKG）和Wnt信号通路。RT-qPCR验证显示，VM组中miR-122-5p表达水平显著低于对照组（<0.05），这与miRNA测序结果一致；VM组中let-7b-5p表达水平显著降低，这与miRNA测序结果相反。双荧光素酶报告基因检测显示，miR-122-5p不能调控或的表达。
母体羊水外泌体中高度丰富的差异表达miRNA可能通过调节MAPK、磷脂酰肌醇-3激酶-蛋白激酶B（PI3K-Akt）、Wnt和cGMP-PKG信号通路在胎儿VM的发生中起重要作用。
