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Introduction Hypertension is the most common complication during pregnancy, affecting 10% of pregnant women worldwide. miRNAs are small, non-coding RNA molecules, which negatively regulate gene expression. They take part in the regulation of placental development and contribute to pregnancy complications, including hypertensive disorders like preeclampsia. miRNAs are released from trophoblast cells to maternal circulation via exosomes predominantly. In preeclampsia, impaired placental function with placental apoptosis and necrosis causes increased release of exosomes. Hsa-mir-210 is one of the so-called hypoximiRs, which are sensitive to hypoxic conditions. Placental hypoxia can be induced by altered placental development and spiral artery remodeling, which are the main characteristics of preeclampsia. Objectives Our aims were to measure the quantity of total-miRNA and to perform expression analysis of exosomal hsa-miR-210 in the circulation of women affected by gestational hypertension (GHT) or preeclampsia (PE). Methods We collected plasma samples from women affected by GHT (n = 10), PE (n = 10) and from healthy controls (n = 10). First, exosomes were isolated using a precipitation method and then miRNAs were extracted from diluted exosomes. We measured the miRNA concentration with Qubit fluorometer applying miRNA specific assay. Reverse transcription was performed by adding UniSp6 spike-in for cDNA synthesis control. RT-PCR was carried out with hsa-miR-210 specific primer and expression was normalized to hsa-miR-103-3p internal control miRNA. The concentrations were calculated using the standard curve method. One-way ANOVA and unpaired t-test were implemented to determine significant differences between the means of the groups. Results The mean total-miRNA concentration was different in the two disease subgroups: 0,29 ± 0,1 ng/ μ l in GHT vs. 0,43 ± 0,15 ng/ μ l in PE, and was significantly smaller in the control group: 0,17 ± 0,07 ng/ μ l, p μ l in GHT, 0,0500 ± 0,0362 ng/ μ l in PE vs. 0,0101 ± 0,0096 ng/ μ l in control , p Conclusions We conclude that the concentration of total-miRNA and exosomal hsa-miR-210 was significantly higher in the circulation of pregnant women affected by hypertension, and it was correlated to the severity of the disease. According to our preliminary results, hsa-miR-210 is a suitable biomarker for hypertensive disorders of pregnancy. |
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