Early-onset preeclampsia, plasma microRNAs, and endothelial cell function
Early-onset preeclampsia, plasma microRNAs, and endothelial cell function
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DOI：
10.1016/j.ajog.2019.11.1286
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发表时间：
2020-05-01
影响因子：
9.8
通讯作者：
Faas, Marijke M.
中科院分区：
文献类型：
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作者：
Lip, Simone V.;Boekschoten, Mark V.;Faas, Marijke M.
背景技术背景：先兆子痫是一种以全身炎症反应和母体内皮功能障碍为病理生理基础的妊娠期高血压疾病。miRNAs是一类非编码小分子RNA，参与基因表达的转录后调控，参与多种生理过程。他们主要下调其靶基因的翻译。目的：我们的目的是比较先兆子痫，健康孕妇和非孕妇的血浆miRNA浓度。此外，我们的目的是评估3个高度增加的血浆中的先兆子痫对内皮细胞功能的影响在vitro.Study设计：我们比较了3391（前体）的血浆样本中的miRNA浓度从早发性先兆子痫的妇女，胎龄匹配的健康孕妇，和非孕妇使用miRNA 3.1。阵列（Affyellow），并通过实时定量聚合酶链反应验证了我们的研究结果。随后，用microRNA模拟物转染内皮细胞（人脐静脉内皮细胞）（我们选择在先兆子痫与健康妊娠中具有最大倍数变化和最低错误发现率的3种miRNA）。转染后，进行功能测定以评估微RNA在内皮细胞中的过表达是否影响体外内皮细胞功能。功能性试验包括伤口愈合试验（测量细胞迁移和增殖）、增殖试验和管形成试验（评估血管生成过程中内皮细胞管的形成）。为了确定miRNA是否能够降低某些基因的基因表达，从转染的内皮细胞中分离RNA，并通过基因表达微阵列（Genechip Human Gene 2.1ST arrays; Life Technologies）评估基因表达（通过测量RNA表达）。对于微阵列，我们使用混合样本，但微阵列中的差异表达基因通过实时定量聚合酶链反应在单个样本中进行验证。
BACKGROUND: Preeclampsia is a hypertensive pregnancy disorder in which generalized systemic inflammation and maternal endothelial dysfunction are involved in the pathophysiology. MiRNAs are small non-coding RNAs responsible for post-transcriptional regulation of gene expression and involved in many physiological processes. They mainly downregulate translation of their target genes.OBJECTIVE: We aimed to compare the plasma miRNA concentrations in preeclampsia, healthy pregnant women, and nonpregnant women. Furthermore, we aimed to evaluate the effect of 3 highly increased plasma miRNAs in preeclampsia on endothelial cell function in vitro.STUDY DESIGN: We compared 3391 (precursor) miRNA concentrations in plasma samples from early-onset preeclamptic women, gestational age-matched healthy pregnant women, and nonpregnant women using miRNA 3.1. arrays (Affymetrix) and validated our findings by real-time quantitative polymerase chain reaction. Subsequently, endothelial cells (human umbilical vein endothelial cells) were transfected with microRNA mimics (we choose the 3 miRNAs with the greatest fold change and lowest false-discovery rate in preeclampsia vs healthy pregnancy). After transfection, functional assays were performed to evaluate whether overexpression of the microRNAs in endothelial cells affected endothelial cell function in vitro. Functional assays were the wound-healing assay (which measures cell migration and proliferation), the proliferation assay, and the tube-formation assay (which assesses formation of endothelial cell tubes during the angiogenic process). To determine whether the miRNAs are able to decrease gene expression of certain genes, RNA was isolated from transfected endothelial cells and gene expression (by measuring RNA expression) was evaluated by gene expression microarray (Genechip Human Gene 2.1 ST arrays; Life Technologies). For the microarray, we used pooled samples, but the differently expressed genes in the microarray were validated by real-time quantitative polymerase chain reaction in individual samples.RESULTS: No significant differences (fold change 1.2 with a false-discovery rate