Elevated microRNA-520g in pre-eclampsia inhibits migration and invasion of trophoblasts.

INTRODUCTION: Pre-eclampsia (PE) is a common and severe obstetric complication. MicroRNAs (miRs) have emerged as molecules that are associated with the disease. METHODS: Quantitative reverse transcription PCR (RT-qPCR) was used for serum miR-520g characterization from 19 severe pre-eclamptic and 19 normal pregnancies. In situ hybridation was adopted to localize microRNA-520g (miR-520g). Migration and invasion of HTR-8/SVneo cells were evaluated after miR-520g mimic treatment with transwell system. MiR-520g target gene was verified in luciferase reporter system. RESULTS: The expression of serum miR-520g displayed an upward trend as pregnancies progress. At first-trimester, miR-520g in pre-eclampsia was significantly higher than that in the control, but no significant differences were found in the second and last trimesters. MiR-520g localized in cytoplasm of early trimester placental trophoblasts. The migration and invasion of HTR8/SVneo were inhibited by miR-520g mimic treatment. Matrix metalloproteinase 2 (MMP2) was verified as a direct target of miR-520g. CONCLUSIONS: Elevated maternal serum level of miR-520g level in first trimester was detected in patients with severe PE. By suppressing the migration and invasion of trophoblast via at least partial inhibition of MMP2 translation inhibition, miR-520g might play a role in the defective spiral artery remodeling, and thus contribute to pre-eclampsia pathophysiology.

Association between the maternal serum levels of 19 eicosanoids and pre-eclampsia.

OBJECTIVE: To investigate whether serum levels of 19 eicosanoids are associated with pre-eclampsia. METHODS: A case-control study was performed using data for pregnant women with pre-eclampsia, normotensive pregnant women, and nonpregnant women, for all of whom serum samples had been collected at a hospital in Shanghai, China, between December 2012 and December 2013. Ultra-performance liquid chromatography-tandem mass spectrometry was used to measure the serum levels of 19 eicosanoids. RESULTS: Overall, 49 pregnant women with pre-eclampsia, 26 normotensive pregnant women, and 14 nonpregnant women were included. Women with pre-eclampsia had significantly higher serum levels of 11,12-epoxyeicosatrienoic acid (11,12-EET), the hydroxyeicosatetraenoic acids 5-HETE, 8-HETE, 12-HETE, and 15-HETE, and leukotriene B4 than did women with a normal pregnancy and nonpregnant women, both before and after the onset of pre-eclampsia (P<0.01 for all comparisons). Women with severe pre-eclampsia had significantly higher serum levels of 5-HETE, 15-HETE, and leukotriene B4 than did women with mild pre-eclampsia, women with a normal pregnancy, and nonpregnant women (P<0.01 for all comparisons). CONCLUSION: The eicosanoids 11,12-EET, 5-HETE, 8-HETE, 12-HETE, 15-HETE, and leukotriene B4 might play important parts in the occurrence and development of pre-eclampsia.

Quantification of preeclampsia-related microRNAs in maternal serum.

To identify the specific serum preeclampsia (PE)-related biomarkers, 10 microRNAs (miRNAs) were selected based on their reported aberrant (4 upregulated and 6 downregulated) expression in PE placenta. A total of 1,035 pregnant patients were enrolled. Finally, 32 pregnancies with PE and 32 healthy pregnancies were incorporated in the study. The expression of these 10 miRNAs in the different trimesters was determined by SYBR-Green reverse transcription-quantitative polymerase chain reaction. Compared with that in the healthy controls, the expression levels of miR-152, miR-183 and miR-210 in PE serum were higher in the second and third trimester, whereas the expression of miR-182 was only higher in the third trimester. The expression levels of 6 miRNAs (miR-1, miR-328, miR-363, miR-377, miR-500 and miR-584) that were downregulated in PE placenta showed no significant differences between pregnancies complicated by PE and healthy pregnancies throughout the 3 trimesters. Areas under the receiver operating characteristic [standard error (SE)] during the 20-24th gestational week for predicting PE were miR-152: 0.94 (SE, 0.026), miR-183: 0.97 (SE, 0.031) and miR-210: 0.93 (SE, 0.018). In conclusion, the expression levels of serum miR-152, miR-183 and miR-210 were elevated since the second trimester in pregnancies complicated with PE, indicating their potentials as serum biomarkers for forecasting PE.

Detection of 19 types of para-arachidonic acids in five types of plasma/serum by ultra performance liquid chromatography-tandem mass spectrometry.

The aim of this study was to examine the consistency of ultra performance liquid chromatography-tandem mass spectrometry (UPLC-TMS) in detecting the levels of para-arachidonic acids (PAAs) among differently processed plasma/serum samples. Ethylenediaminetetraacetic acid (EDTA)-K2, sodium citrate, heparin lithium, coagulant/separation gel, and coagulant-free vacuum blood-sampling tubes were used to collect the fasting blood samples from 15 volunteers. All blood samples were subjected to solid-phase extraction using an Oasis HLB µElution 96-well plate, and UPLC-TMS was used to detect 19 types of PAAs in the blood samples. Within the plasma samples, the concentrations of 5, 6-DHET; 11, 12-epoxyeicosatrienoic acid (EET); 5-hydroxyeicosatetraenoic acid (HETE); leukotriene B4 (LTB4); plasma thromboxane B2 (TXB2); and 12-HETE were significantly higher in the heparin lithium group than in the EDTA-K2 and sodium citrate groups. Within the serum samples, the concentration of LTB4 was significantly higher in the coagulant/separation gel group than in the coagulant-free group, while that of TXB2 was significantly higher in the coagulant-free group than in the coagulant/separation gel group. The levels of some types of PAAs in differently processed plasma/serum samples were inconsistent, and the concentrations of 5, 6-DHET; 5-HETE; 12-HETE; TXB2; and LTB4 were significantly higher in the two serum samples and the heparin lithium group than in the EDTA-K2 and sodium citrate groups.

Effect of 20-hydroxyeicosatetraenoic acid on biological behavior of human villous trophoblasts and uterine vascular smooth muscle cells.

During pregnancy, disorders in uterine spiral artery remodeling (USAR) cause preeclampsia and other diseases. The aim of this study was to investigate the effect of 20-hydroxyeicosatetraenoic acid (20-HETE) on the biological behavior of human villous trophoblasts (HVTs) and human uterine vascular smooth muscle cells (HUVSMCs), and explore the role of 20-HETE in USAR. 20-HETE and its inhibitor HET0016 were used to study migration, invasion and apoptosis in the HVT and HVT-HUVSMC models. 20-HETE inhibited the migration and invasion of HVTs, and inhibited apoptosis in HUVSMCs and HUVSMCs co-cultured with HVTs. 20-HETE had thus obvious effects on the biological behavior of HVTs and HUVSMCs. These effects may cause USAR disorders and vascular dysfunction, leading to preeclampsia.