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).
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.
The mean total-miRNA concentration was different in the two disease subgroups: 0,29 ± 0,1 ng/223a3142e9e584c26c6fa6d16c3af231" title="Click to view the MathML source">μl in GHT vs. 0,43 ± 0,15 ng/223a3142e9e584c26c6fa6d16c3af231" title="Click to view the MathML source">μl in PE, and was significantly smaller in the control group: 0,17 ± 0,07 ng/223a3142e9e584c26c6fa6d16c3af231" title="Click to view the MathML source">μl, p < 0,05. We found that exosomal hsa-miR-210 was significantly upregulated in the circulation of pregnant women affected by hypertensive disorders compared to the normotensive pregnant women: 0,0357 ± 0,019 ng/223a3142e9e584c26c6fa6d16c3af231" title="Click to view the MathML source">μl in GHT, 0,0500 ± 0,0362 ng/223a3142e9e584c26c6fa6d16c3af231" title="Click to view the MathML source">μl in PE vs. 0,0101 ± 0,0096 ng/223a3142e9e584c26c6fa6d16c3af231" title="Click to view the MathML source">μl in control , p < 0,05.
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.