sPE Patients Research Articles

MiR-139-5p promotes the proliferation and invasion of trophoblast cells by targeting sFlt-1 in preeclampsia

BackgroundThe biological functions of placental trophoblast cells have been reported to be critical in preeclampsia (PE) and its complications. Here, we aimed to investigate the role and underlying mechanism of soluble fms-like tyrsine kinase-1 (sFlt-1) and miR-139-5p in severe preeclampsia (sPE) by culturing the trophoblast cells from patients. MethodsELISA and qRT-PCR were used to measure the expression of sFlt-1 and miR-139-5p. The direct interaction between sFlt-1 and miR-139-5p was determined by luciferase reporter assay. Cell proliferation and invasion were evaluated by CCK-8 analysis and transwell assay. ResultsOur results showed that miR-139-5p was downregulated in sPE patients and was negatively correlated with the expression of sFlt-1. Further, sFlt-1 was a direct target of miR-139-5p, which monitored the expression of sFlt-1. Besides, miR-139-5p promoted the proliferation and invasion of trophoblast cells derived from sPE patients. Overexpression of sFlt-1 attenuated the effects of miR-139-5p on cell proliferation and invasion of trophoblast cells from sPE patients. ConclusionOur research proposes a novel mechanism where the role of miR-139-5p is dependent on sFlt-1. Our data demonstrated that miR-139-5p promoted the proliferation and invasion of trophoblast cells by directly targeting sFlt-1 in PE.

The expression profile of circRNA and its potential regulatory targets in the placentas of severe pre-eclampsia

ObjectiveTo determine the expression profiles of circular RNAs (circRNAs) of women with severe pre-eclampsia (sPE group) versus normal pregnancies (normal control group). Materials and methodsRNA-sequencing (RNA-seq) was conducted to characterize differentially expressed circRNAs and mRNAs in the placental tissues of women with sPE versus normal pregnancies. circRNA functions were predicted by Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) database analysis. The backsplicing junctions of circRNAs were validated with the use of divergent primers. Relative expression levels of cirRNAs were verified by quantitative real-time PCR (qPCR). A circRNA-miRNA-mRNA interaction network was constructed to outline the regulatory network of the differentially expressed circRNAs. ResultsA total of 49 differentially expressed circRNAs were found in the placental tissues of women with sPE. Several differentially expressed mRNAs were also observed in the sPE patients. KEGG analysis revealed that the most enriched pathway of the circRNAs was the MAPK signaling pathway, while the differentially expressed mRNAs were primary enriched in pathways in cancer. Among these circRNAs, hsa_circ_0001438, hsa_circ_0001326, and hsa_circ_32340 were upregulated in the sPE patients and the circRNA-miRNA-mRNA interaction network generated with these three circRNAs revealed a broad regulatory network that might be involved in the pathogenesis of sPE. ConclusioncircRNAs are differentially expressed in sPE. The upregulation of hsa_circ_0001438, hsa_circ_0001326, and hsa_circ_32340 has a potential role in the regulation of miRNA and mRNA expression. Changes to the expression profiles of the circRNAs might be linked to the pathogenesis of sPE and could function as biomarkers.

Mme V. Grossesse délirante en pré-ménopause

Small ubiquitin-like modifiers (SUMO) conjugate to target proteins in a dynamic, reversible manner to function as post-translational modifiers. SUMOylation of target proteins can impinge on their localization, in addition to their activity or stability. Differential expression of deSUMOylating enzymes (SENP 1 and 2) contributes to altered mammalian placental development and function in mice. Severe preeclampsia (sPE) is associated with abnormal placental development and chronic ischemic injury. Extra- and intracellular stimuli/stressors that include hypoxic-activated pathways are known modulators of SUMOylation. In this current study we hypothesized that placentas from sPE patients will display up regulation in the SUMO regulatory pathway.Utilizing qRT-PCR, immuno-blotting and Western techniques, we determined the expression levels of SUMO pathway genes in healthy and diseased placentas. We also exposed placental explants to hypoxia to study the effect on the SUMOylation pathway.We observed steady-state expression of SUMO1–3, SUMO-conjugated enzyme-UBC9 and deSUMOylating enzymes – SENPs, throughout normal gestation. An elevated level of free SUMO1–3 and SUMO-protein conjugates was observed in sPE placentas. Furthermore, placental UBC9 levels were strikingly increased in the same sPE patients. Hypoxia-induced SUMOylation in first trimester placental explants.Our data demonstrate an elevated steady-state of SUMOylation in sPE placentas compared with gestational aged-matched controls. The observed hyper-SUMOylation in sPE placentas correlates with elevated expression of UBC9 rather than with reduced expression of SENPs Hypoxia may contribute to alterations in placental SUMOylation pathway.Increased placental SUMOylation may contribute to the pathogenesis of serious placental pathology that causes extreme preterm birth.

How do the mechanical influences of blood flow impact trophoblast plugging and unplugging of the spiral arteries in early pregnancy?

The biological functions of placental trophoblast cells have been reported to be critical in preeclampsia (PE) and its complications. Here, we aimed to investigate the role and underlying mechanism of soluble fms-like tyrsine kinase-1 (sFlt-1) and miR-139-5p in severe preeclampsia (sPE) by culturing the trophoblast cells from patients.ELISA and qRT-PCR were used to measure the expression of sFlt-1 and miR-139-5p. The direct interaction between sFlt-1 and miR-139-5p was determined by luciferase reporter assay. Cell proliferation and invasion were evaluated by CCK-8 analysis and transwell assay.Our results showed that miR-139-5p was downregulated in sPE patients and was negatively correlated with the expression of sFlt-1. Further, sFlt-1 was a direct target of miR-139-5p, which monitored the expression of sFlt-1. Besides, miR-139-5p promoted the proliferation and invasion of trophoblast cells derived from sPE patients. Overexpression of sFlt-1 attenuated the effects of miR-139-5p on cell proliferation and invasion of trophoblast cells from sPE patients.Our research proposes a novel mechanism where the role of miR-139-5p is dependent on sFlt-1. Our data demonstrated that miR-139-5p promoted the proliferation and invasion of trophoblast cells by directly targeting sFlt-1 in PE.

Downregulation of miR-424 in placenta is associated with severe preeclampsia

Previous studies have suggested that altered miRNA expression in the placenta is associated with preeclampsia. The aim of this study was to investigate the expression of miR-424 in placental samples of severe preeclampsia (sPE) and uncomplicated pregnancy patients. miRNA was isolated from placentas obtained from 30 sPE patients and 30 healthy women. Quantitative real-time polymerase chain reaction was used to analyze the expression of miR-424. The prediction of target genes of miR-424 was performed using miRGen database. The function of these target genes was analyzed further by DAVID and Gorilla software. The expression of miR-424 was significantly lower in patients with sPE than in healthy controls. Changed expression of miR-424 in the case of pregnancy-related hypertensive disorders might affect the Wnt signaling pathway. These factors have a strong correlation with the development of PE. Expression of miR-424 in placenta was lower in patients with sPE, suggesting its role in the pathology of sPE.

Various levels of circulating exosomal total-miRNA and miR-210 hypoxamiR in different forms of pregnancy hypertension.

Hypertension is a common complication during pregnancy, affecting 10% of pregnant women worldwide. Several microRNA (miRNA) were shown to be involved in hypertensive disorders of pregnancy. In preeclampsia (PE), placental dysfunction causes the enhanced release of extracellular vesicle-derived miRNAs. The hypoxia-sensitive hsa-mir-210 is the most common PE-associated miRNA, but its exosomal profile has not been investigated. Our aims were to measure exosomal total-miRNA concentration and to perform expression analysis of circulating exosomal hsa-miR-210 in women affected by chronic hypertension (CHT) gestational hypertension (GHT) or PE. We collected plasma samples from women with CHT, GHT, PE (moderate: mPE and severe: sPE) and from normotensive pregnancies. Exosomal miRNAs were extracted and miRNA concentration was measured. RT-PCR was carried out with hsa-miR-210-3p-specific primers and relative expression was calculated using the comparative Ct method. The total-miRNA concentration was different in the disease subgroups, and was significantly higher in mPE and sPE compared to the other groups. We found a significant difference in the relative exosomal hsa-miR-210-3p expression between all hypertensive groups compared to the normotensive samples, but significant upregulation was only observed in case of mPE and sPE patients. Both the level of total-miRNA and hsa-miR-210 expression was higher in case of severe PE. The level of circulating exosomal total-miRNA and hsa-miR-210 was elevated in women with PE, and it was higher in the severe form. We showed that hsa-miR-210 is secreted via exosomes, which may have a role in the pathomechanism of the disease.

Expression profiling of maternal plasma and placenta microRNAs in preeclamptic pregnancies by microarray technology

Preeclampsia (PE) is one of the leading causes of maternal and fetal morbidity and mortality, occurring usually in the second half of pregnancy and affecting approximately 5–8% of pregnancies in the world. miRNAs play critical role in the regulation of placental development processes. We aimed to determine specific novel miRNAs for early diagnosis of preeclampsia which is one of the most dangerous pregnancy diseases. In this study 72 samples, maternal age 22 ≤ and ≤36, have been analyzed; maternal plasma and placental miRNAs were isolated from 18 severe preeclampsia (sPE) patients and 18 controls, respectively. Profiling of human miRNAs (1368 probe) was performed in samples with Agilent v16 microarrays for detection of the differences in miRNA expression between two groups. The results were validated by using TaqMan RT-qPCR method. The analysis indicated that 406 of these miRNAs in all placentas and 42 of these miRNAs in all maternal plasma were expressed. The relative expression analysis has shown that 12 miRNAs (p < 0.05 and >2-fold) in maternal plasma were differentially expressed in PE and control group. However, five miRNAs were validated by qRT-PCR. Once validated miRNAs have been searched in databases for their target genes and function, it has been shown that there are some preeclampsia related pathways as a target such as angiogenesis, cardiovascular, hypertension, placental abruption and preeclampsia disorders. Differentially expressed and validated plasma miRNAs might be used as notable biomarkers for non-invasive early diagnosis of preeclampsia and treatment of disease.

Increased circulating Th22 cells correlated with Th17 cells in patients with severe preeclampsia

ABSTRACTObjective: We aimed to investigate Th22 cells and their association with Th17 and Treg cells in the etiology of severe preeclampsia (sPE).Methods: Thirty sPE patients and 30 healthy pregnant women were recruited in this study. The percentages of Th17, Th22, and regulatory T cells (Tregs) in the peripheral blood were measured by flow cytometry. ELISA was used to measure the plasma concentrations of interleukin (IL)-17, IL-22, and IL-10.Results: The percentages of Th17 and Th22 cells and the plasma concentrations of IL-17 and IL-22 were significantly increased in sPE patients along with a decreased percentage of Treg cells and a decreased plasma IL-10 concentration. There was a positive correlation between the levels of Th22 cells and Th17 cells in sPE patients. Moreover, a positive correlation was found between plasma IL-22 concentration and the percentage of Th22 cells in sPE patients.Conclusions: Increased circulating Th22 cells, which were correlated with Th17 cells, were observed in patients with sPE. The immune imbalance between T helper (Th) cells may contribute to the pathogenesis of sPE.

Assessment of ADMA, estradiol, and progesterone in severe preeclampsia

ABSTRACTBackground: Decreased nitrous oxide (NO) levels are crucial factors in severe preeclampsia (sPE), and asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of NO synthetase. Steroid hormones are closely related to the vascular endothelium. This study determined the levels of and correlations between ADMA, estradiol (E2), and progesterone (Pg) in sPE to investigate the roles of these factors in this disease. Methods: Sixty-two sPE patients (sPE group) were divided into the sPE1 subgroup (28+1–32+0 weeks of pregnancy), the sPE2 subgroup (32+1–36+0 weeks), and the sPE3 subgroup (36+1–40+0 weeks) and 75 normal pregnant women (NC group) were divided into the NC1 subgroup (28+1–32+0 weeks of gestation), the NC2 subgroup (32+1–36+0 weeks), and the NC3 subgroup (36+1–40+0 weeks). Serum and placental ADMA levels were measured by enzyme-linked immunosorbent assay (ELISA), and serum E2 and Pg concentrations were determined by the chemilumineseent immunoassay (CLIA). Results: ADMA concentrations in both the placenta and the maternal serum were significantly higher in the sPE group (p < 0.05). Higher ADMA contents were observed in the placenta relative to the maternal serum (p < 0.05). Serum E2 levels were significantly lower in the sPE group (p < 0.05). For Pg, the only significant difference was observed between the sPE1 and NC1 subgroups (p < 0.05). The Pg/E2 ratios in the sPE groups were significantly higher, with a significant high positive correlation between Pg/E2 ratios and serum ADMA levels. Conclusion: Increased serum levels of ADMA in sPE may result from increased secretion from the placenta, and the increased Pg/E2 ratio may play a role in the development of sPE by aggravating ADMA.

Variations of MicroRNAs in Human Placentas and Plasma From Preeclamptic Pregnancy

Preeclampsia is a major cause of maternal and fetal mortality and morbidity worldwide. The differential expression of several microRNAs (miRNAs) has been found in preeclamptic placentas. However, great conflict exists regarding this aspect, and detailed examinations have largely been lacking of miRNA profiles in different parts of the placenta and in maternal plasma of women with this disorder. In this study, a total of 9 downregulated miRNAs (miR-195, miR-223, miR-218, miR-17, miR-18a, miR-19b1, miR-92a1, miR-379, and miR-411) and 7 upregulated miRNAs (miR-210, miR-30a-3p, miR-518b, miR-524, miR-17-3p, miR-151, and miR-193b) were identified in severe preeclampsia (sPE) placentas when compared with normal pregnant controls. In addition, sampling position in the chorionic or basal plate of placenta led to evident variations in differential miRNAs of sPE placentas. In a prospective pregnant cohort, we found that the circulating levels of 3 members of miR-17-92 cluster (ie, miR-18a, miR-19b1, and miR-92a1) were significantly lower, whereas that of miR-210 was higher in sPE patients than those in normal controls at gestational weeks 15 to 18 and at term. The results of in situ hybridization revealed the localization of miR-18a, miR-92a1, and miR-210 in various subtypes of placental trophoblasts and endothelial cells. In human trophoblast cell line, HTR8/SVneo cells, miR-18a could promote trophoblast cell invasion via targeting and suppressing Smad2 expression. This study provides fundamental evidences for exploring the roles of miRNAs in the pathogenesis of preeclampsia.

Emerging role of SUMOylation in placental pathology

IntroductionSmall ubiquitin-like modifiers (SUMO) conjugate to target proteins in a dynamic, reversible manner to function as post-translational modifiers. SUMOylation of target proteins can impinge on their localization, in addition to their activity or stability. Differential expression of deSUMOylating enzymes (SENP 1 and 2) contributes to altered mammalian placental development and function in mice. Severe preeclampsia (sPE) is associated with abnormal placental development and chronic ischemic injury. Extra- and intracellular stimuli/stressors that include hypoxic-activated pathways are known modulators of SUMOylation. In this current study we hypothesized that placentas from sPE patients will display up regulation in the SUMO regulatory pathway. MethodsUtilizing qRT-PCR, immuno-blotting and Western techniques, we determined the expression levels of SUMO pathway genes in healthy and diseased placentas. We also exposed placental explants to hypoxia to study the effect on the SUMOylation pathway. ResultsWe observed steady-state expression of SUMO1–3, SUMO-conjugated enzyme-UBC9 and deSUMOylating enzymes – SENPs, throughout normal gestation. An elevated level of free SUMO1–3 and SUMO-protein conjugates was observed in sPE placentas. Furthermore, placental UBC9 levels were strikingly increased in the same sPE patients. Hypoxia-induced SUMOylation in first trimester placental explants. DiscussionOur data demonstrate an elevated steady-state of SUMOylation in sPE placentas compared with gestational aged-matched controls. The observed hyper-SUMOylation in sPE placentas correlates with elevated expression of UBC9 rather than with reduced expression of SENPs Hypoxia may contribute to alterations in placental SUMOylation pathway. ConclusionIncreased placental SUMOylation may contribute to the pathogenesis of serious placental pathology that causes extreme preterm birth.

Circulating microRNAs are elevated in plasma from severe preeclamptic pregnancies

Until recently, the molecular pathogenesis of preeclampsia (PE) remained largely unknown. Reports have shown that circulating microRNAs (miRNAs) are promising novel biomarkers for cancer, pregnancy, tissue injury, and other conditions. The objective of this study was to identify differentially expressed miRNAs in plasma from severe preeclamptic pregnancies compared with plasma from normal pregnancies. By mature miRNA microarray analysis, 15 miRNAs, including 13 up- and two downregulated miRNAs, were screened to be differentially expressed in plasma from women with severe PE (sPE). Seven miRNAs, namely miR-24, miR-26a, miR-103, miR-130b, miR-181a, miR-342-3p, and miR-574-5p, were validated to be elevated in plasma from severe preeclamptic pregnancies by real-time quantitative stem-loop RT-PCR analysis. Gene ontology and pathway enrichment analyses revealed that these miRNAs were involved in specific biological process categories (including regulation of metabolic processes, regulation of transcription, and cell cycle) and signaling pathways (including the MAP kinase signaling pathway, the transforming growth factor-β signaling pathway, and pathways in cancer metastasis). This study presents, for the first time, the differential expression profile of circulating miRNAs in sPE patients. The seven elevated circulating miRNAs may play critical roles in the pathogenesis of sPE, and one or more of them may become potential markers for diagnosing sPE.

Effect of lipoxin A4 on interleukin-1β production of human monocytes from severe preeclamptic women and its relationship with cytoplasm free calcium concentration-an in vitro study

Objective To investigate the effect of lipoxin A4(LXA4) on interleukin-1β(IL-1β)production of monocytes in maternal peripheral blood from severe preeelampsia women and its relationship with cytosolic free calcium([Ca2+]i)concentration. Methods Peripheral venous blood was drawn from 15 women with severe preeelampsia(SPE group)and 20 normal pregnant women (control group)who were admitted to the First Affiliated Hospital of Wenzhou Medical College from October 2008 to May 2009.Monoeytes were obtained from peripheral blood with Ficolt density gradient centrifugation and then were suspended in RPMI-1640 culture supplemented with LXA4 at the final concentrations of 0,10 and 100 nmol/L,respectively.IL-1β levels in monocytes supernatant were detected by enzyme-linked immunosorbent assay.The cytoplasma[Ca2+]i of cultured monocytes and its variations affected by LXA4 were measured by laser scanning confocal microscope. Results (1) After incubation with different concentrations of LXA4(0,10,100 nmol/L)for 24 h,the levels of IL-1β in SPE group were(63.16±8.20)pg/L,(53.71±8.08)pg/L and(43.16±6.89)pg/L,respectively, indicating a significant inhibition effect on IL-1β level in a dose-dependent manner (P 0.05). (2) Without LXA4, the [Ca2+ ]i concentrations of monocytes in the SPE group were higher than that of the control (1028.09 ± 160. 52 vs 323.61 ±87.86, P<0. 05). After treatment with 100 nmol/L LXA4, the [Ca2+ ]i concentration of monocytes significantly decreased in the SPE group (409.67± 116.73, P<0. 05). Conclusions In vitro LXA4 may inhibit the IL-1β production in monoeytes of SPE patients through decrease of the cytoplama [Ca2+ ]i concentration. Key words: Lipoxins; Pre-eclampsia; Monocytes; Interleukin-lbeta; Calcium