sFlt-1 Concentrations Research Articles

Outcomes of pregnancies in women with extremely high ratio of sFlt-1 / PlGF: a series of clinical cases

Preeclampsia as one of the manifestations of placental dysfunction associated with dysregulation of pro- and antiangiogenic factors. During normal pregnancy, the concentration of antiangiogenic factor sFlt-1 remains low, which allows accurate transmission of signals induced by proangiogenic factors VEGF and PlGF. This balance is crucial to maintain physiological vasodilation. Under hypoperfusion, the placenta increases the synthesis of sFlt-1, trying to increase maternal pressure and increase placental perfusion.The objective: to study the clinical outcomes of pregnancies in a group with extremely high ratios of sFlt-1 and PlGF.Materials and methods. The results of determining the levels of placental growth factor (PlGF) and soluble fms-like tyrosine kinase (sfLT-1), which were performed during 2017-2020 in the serum of 128 pregnant women at 18–39 weeks, were analyzed.Results. The mean age of pregnant women was 28,4±4 years. The average gestational age during the study was 26±5 weeks. The majority of pregnant women had signs of IUGR (62,5%) and/or preeclampsia (50%) during the study. The average level of the sFlt/PIGF ratio was 1747 pg/ml. In 100% of cases with a digital ratio of more than 850, signs of an obstetric angiogenic catastrophe that required immediate delivery developed during the nearest future. The average time from analysis to delivery was 10 days. In 50% of pregnant women, urgent delivery was required within 48 hours after analysis. Favorable neonatal results were observed in 37,5% of women.Conclusion. We found a tendency for worsening perinatal outcomes in women with extremely high sFlt-1/PlGF ratios after reaching 850. Careful monitoring of the pregnant woman and consideration of need for prophylactic administration of corticosteroids to accelerate fetal lung maturation in this group is rational.

1072 Early pregnancy BMI and angiogenesis-related biomarkers (sFlt-1/PlGF): a pathway for adverse pregnancy outcomes?

Obesity is a risk factor for adverse pregnancy outcomes, but it is unclear whether this is mediated through placental angiogenic factors. We examined the relationship between early pregnancy body mass index (BMI) and 1st- and 2nd-trimester concentrations of sFlt-1/PlGF as a potential biologic pathway for placental insufficiency. We conducted secondary analyses of data and samples from a large, multi-site prospective cohort study of nulliparous pregnant women conducted in the U.S. First- and 2nd-trimester blood samples, 1st-trimester BMI, demographic, lifestyle, and pregnancy data were gathered for all participants. sFlt-1 and PlGF concentrations were measured for a subsample of 1,502 cases (delivery<37weeks, preeclampsia or eclampsia, birthweight for gestational age <5th%, or stillbirth) and 911 controls. We used one-way ANOVA and multivariable linear and logistic regression models to test the association of BMI and sFlt-1/PlGF ratio, using a previously published ratio cutoff of ≥38 in logistic regression. We identified potential confounders a priori and used backward elimination to determine the best-fit model. Plots of mean sFlt-1/PlGF ratios by BMI category and gestational week showed lower ratios in overweight and obese groups in the 1st trimester, which reversed to higher ratios among overweight and obese groups in the 2nd trimester (Fig. 1). After controlling for clinical and demographic factors, women in all high BMI groups had significantly lower mean sFlt-1/PlGF ratios than normal weight women in the 1st trimester (Table 1). In the 2nd trimester, women with BMI ≥35 had a 35% higher mean sFlt-1/PlGF ratio than normal weight women (p=0.009). Logistic regression suggested increased odds of a 2nd-trimester ratio ≥38 among women with BMI ≥35, though not statistically different from 1.0 (OR 2.59, 95% CI 0.95-7.08). Early pregnancy BMI is associated with alterations in first- and second-trimester sFlt-1/PlGF ratios. Findings suggest a potential physiologic pathway between obesity and placental insufficiency.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

27 Metformin to treat Preterm Pre-eclampsia (PI-2): A randomised, double blind, placebo-controlled trial

Preterm preeclampsia has high rates of maternal and perinatal morbidity and mortality. No drugs have clearly shown a prolongation in gestation. Preclinical studies have identified metformin as a potential treatment. We performed a double-blind, randomised placebo-controlled trial at Tygerberg Hospital, South Africa. We randomly assigned women with preterm preeclampsia between 26+0 to 31+6 weeks gestation and undergoing expectant management to 3 grams of oral metformin XR (extended released) in divided doses, or placebo. The primary outcome was prolongation of pregnancy. Secondary outcomes included maternal and neonatal composite, and individual outcomes. Before the trial we performed metformin pharmacokinetic studies on 15 women with preterm preeclampsia. Between Feb 2018 and Mar 2020 we randomly allocated 180 participants to receive metformin (n=90) or placebo (n=90). Pharmacokinetic studies demonstrated oral metformin XR resulted in good circulating drug concentrations. The primary outcome analysis included 87 allocated to metformin and 84 assigned to placebo. The median time from randomization to delivery was 16.2 days (interquartile range (IQR), 5.2-28.8 days) in the metformin group and 9.5 days (IQR 3.4-22.7) days in the placebo group (difference of 6.7 days (95% Confidence Interval (CI) -0.17 to 13.64; P=0.056; Fig 1). In a pre-specified per protocol analysis that included only those who were compliant, the median prolongation was 16.2 (IQR 5.3-27.5; n=75) versus 7.4 (IQR 2.9-21.2; n=72) days, respectively (difference of 8.4 days, 95%CI 1.0-15.8; P=0.026). There were trends towards increased birthweight (1538g vs 1402g, difference of 136g, 95% CI-12 to 285;P=0.07) and a decreased length of stay (12 vs 18.0, difference of 6 days (95% CI-0.4 to 12.4;P=0.07) in the neonatal nursery among those in the metformin group (Table 1). There were no differences in composite maternal or neonatal outcomes or circulating concentrations of sFlt-1 or placental growth factor. In women with preterm preeclampsia, metformin prolongs gestation and may improve neonatal outcomes.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Association study of hypertension susceptibility genes ITGA9, MOV10, and CACNB2 with preeclampsia in Chinese Han population

Objective Preeclampsia (PE) is a disorder that occurs during the pregnancy and could affect the maternal and perinatal mortality as well as morbidity. The aim of our study is to investigate the associations between the hypertension susceptibility genes ITGA9, MOV10 and CACNB2 with PE in Chinese Han population. Methods A case–control study including 178 PE patients and 202 healthy controls was conducted to assess the associations between three loci (ITGA9 rs155524, MOV10 rs2932538 and CACNB2 rs4373814) and PE. The TaqMan probe assay was applied for genotyping in our study. Quantitative real-time PCR was performed to detect the mRNA expression levels of ITGA9, MOV10 and CACNB2. ELISA was carried out to detect the concentration of serum sFlt-1 or PLGF. Results Our study detected no significant differences in allelic frequencies of three SNPs between PE patients and healthy controls. In the genetic model, the results showed that the patients with ITGA9 rs155524 GA or AA genotypes had a higher risk of PE development compared to those with GG genotype in codominant model. And PE patients had a higher frequency of GA + AA genotypes based on the dominant model. Subgroup analysis showed ITGA9 rs155524 was associated with early-onset PE but not with late-onset PE. No association was observed between MOV10 and CACNB2 with PE in any genetic model and subgroup analysis. Quantitative real-time PCR results showed that ITGA9 mRNA expression level was apparently increased in the placental tissues of PE patients. In addition, ITGA9 expression levels of GA + AA subjects were apparently higher than that in the genotype GG of placental tissues. sFlt-1/PLGF ratio was higher in GA + AA subjects than that in GG subjects. Regression analysis revealed that ratio of sFlt-1/PLGF was positively correlated with ITGA9 mRNA expression level. Conclusion This study has identified ITGA9 is a promising candidate susceptibility gene for early-onset PE. Our findings demonstrated that the high expression of ITGA9 might be associated with an increased risk of PE.

Increasing efficiency of preterm birth prediction

Abstract. The disturbed placentation in a certain way plays an important role in the pathogenesis of great obstetric syndromes. Therefore, the possibility of using biochemical indicators of the level of placental growth factor (PlGF) and fms-like tyrosine kinase (sFlt-1) for predicting preterm labor is of great interest. The aim of the study was developing criteria for predicting preterm labor based on the combined use of ultrasound cervicometry and the concentration of PlGF and sFlt-1. Materials and methods. A total of 227 pregnant women were examined, 190 of whom had preterm birth. Group I included 48 women whose pregnancy completed at 23 to 27 weeks. In group II, 142 women with prematurity in terms of 28 to 36 weeks were observed. Group III included 37 women with a healthy pregnancy, which completed at 38–41 weeks. All patients involved in the study underwent ultrasound cervicometry on a Voluson 730 (GE Healthcare, USA) at 16 weeks, and also the concentration of PlGF and sFlt-1 in blood serum by electrochemiluminescence immunoassay using a Cobas e411 analyzer was studied (Roche diagnostics, Switzerland). Results. The study revealed a significant (p &lt; 0.001) decrease in the average concentration of PlGF in the blood serum by 65 % associated with an increased level of sFlt-1 by 93% in women with early termination of pregnancy – 23–27 weeks (group I) in relation to women with a healthy pregnancy. In women with preterm birth at 28–36 weeks of gestation (group II), a similar trend of changes was observed: the PlGF level significantly (p = 0.014) decreased on average by 68 % in relation to women with a healthy pregnancy. In women of group I, the value of the sFlt-1 / PlGF ratio exceeded (p &lt; 0.001) the values of patients in group III by 14 times. But an even more pronounced imbalance of angiogenic factors was observed in women of group II – on average 16 times. Two main markers were used to predict premature birth: shortening of the cervix according to ultrasound cervicometry data of less than 30 mm, as well as the sFlt-1/PlGF ratio of more than 50 conv. units. The odds ratio (OR) of preterm birth in the case of using only the length of the cervix was 34,133 (95 % confidence interval – 12.308–94.660). OR in the case of combined use of cervicometry data and angiogenesis variables was 148.750 (95 % confidence interval – 33.243–665.593). Conclusion. Thus, the use of an additional biochemical marker significantly improved the accuracy of preterm labor prediction.

Effects of modified lipoproteins on human trophoblast cells: a role in pre-eclampsia in pregnancies complicated by diabetes

IntroductionPre-eclampsia (PE) is increased ~4-fold by maternal diabetes. Elevated plasma antiangiogenic factors, soluble fms-like tyrosine kinase (sFLT-1) and soluble endoglin (sENG), precede PE onset. We investigated whether diabetes-related stresses, modified...

Antiangiogenic Biomarker Soluble Fms-like Tyrosine Kinase-1 in Pregnancy Complicated with Preeclampsia: A Cohort Study

Introduction: Preeclampsia (PE) is one of the leading obstetric diseases with significant morbidity and mortality in both mother and foetus. The etiology of preeclampsia is unknown. It may result from several reasons with imbalance between angiogenic regulatory factors in maternal circulation as one of the factor. High circulatory levels of Soluble Fms-like Tyrosine Kinase-1 (sFlt-1) are detectable several weeks before clinical presentation of preeclampsia. Aim: To determine association of serum levels of sFlt-1 with preeclampsia in second and third trimester of pregnancy. Materials and Methods: A prospective cohort study was conducted in the Department of Biochemistry in collaboration with the Department of Obstetrics and Gynaecology in a tertiary care hospital (Sri Guru Ram Das Institute of Medical Sciences and Research, Amritsar, Punjab, India), from November 2018 to March 2021. The study participants were divided into two groups i.e, normotensive (group 1) and preeclamptic (group 2). The enrollment of participants was done during second trimester. Serum sFlt-1 concentration was measured in second trimester (24-28 weeks) and in third trimester (beyond 28 weeks) using Enzyme Linked Immunoassay (ELISA) kits. Receiver Operating Characteristics (ROC) curve analysis was done for evaluation of Area Under Curve (AUC), sensitivity and specificity using software defined cut-off values. Results: Total 60 participants were there in each group with maternal mean age of 25.8±3.2 years in group 1 and 30.6±5.5 years in group 2 (p-value &lt;0.001). sFlt-1 levels were significantly higher in preeclampsia group both in second and third trimesters when compared with normotensive group with median 313.07 versus 65.150 (p-value &lt;0.001) and 337.875 versus 76.925 (p-value &lt;0.001), respectively. The ROC curve analysis using 190.5 ng/mL as cut-off point in second trimester showed sensitivity 90%, specificity 85%, AUC was 0.832, 95% CI (0.745-0.918) and in third trimester at cut-off point 271.5 ng/mL showed sensitivity 90%, specificity 90%, AUC was 0.884, 95% CI (0.817-0.951). Conclusion: The soluble Fms-Like Tyrosine Kinase-1-1 (sFlt-1), may serve as biomarker for early diagnosis and can improve prediction of preeclampsia.

Are serum Netrin-4 levels predictive of preeclampsia?

To investigate the levels of anti-angiogenic factors, namely sFlt-1 and Netrin-4, in patients with preeclampsia (PE). Cord-blood (UC) sFlt-1 and Netrin-4 concentrations were measured in 30 patients with severe PE, 30 patients with PE and 30 control infants and their mothers (MS). Maternal sFlt-1 levels were significantly higher in the severe PE and PE groups than in the control group. There were no statistical differences among the three groups in maternal and fetal Netrin-4 levels. But Netrin-4 levels were found to be the lowest in the control group and higher in the PE and severe PE groups. The correlation analysis revealed a positive correlation between maternal sFlt-1 levels and maternal Netrin-4 levels (p = 0.012, and r = 0.263), maternal sFlt-1 levels and fetal sFlt-1 levels (p = 0.012, and r = 0.263). There was a positive correlation found between maternal sFlt-1 levels and maternal Netrin-4 levels. We are of the opinion that elevation in levels of Netrin-4 might be secondary to placental hypoxia occurring in PE. The present study led to the consideration of anti-angiogenic biomarkers (sFlt-1 and Netrin-4) on automated platforms for clinical use as an aid in establishing the diagnosis and prognosis of PE.

Association between urinary placental protein 13 and soluble fms-like tyrosine kinase-1 in preeclamptic women in the third trimester of pregnancy

Background: Preeclampsia (PE) is diagnosed after 20 weeks of gestation. This multisystem disorder affects 2-7% of pregnant women. PE is a serious complication of pregnancy and one of the main causes of maternal and neonatal mortality and morbidity in the world. The inadequate placentation process results in a change in anti-angiogenic factors levels, such as placental protein 13 (PP13) and soluble fms-like tyrosine kinase 1 (sFlt-1). Objectives: To investigate the correlation between urinary PP13 and sFlt-1 in preeclamptic women in their third trimester of pregnancy. Methods: A case-control study was carried out from August 2018 till January 2019. Urine samples were collected from pregnant women at Al-Elweyia Hospital, Al-Hakeem Hospital, and Al- Imamain Al-Kadhimain Medical City (in Baghdad, Iraq). The patients’ group include fifty women with PE in the third trimester (25 mild and 25 severe). Fifty healthy pregnant women (at their third trimester of pregnancy) were studied as a control group. Results: The mean urinary PP13 levels were decreased in women with PE significantly (p≤ 0.05) (mild and severe) compared with healthy women (43.44±4.914 pg/ml, 33.34±1.863 pg/ml, and 51.84 ±2.60 pg/ml) respectively. Also, urinary sFlt-1 concentrations were decreased non-significantly (p&gt; 0.05) in women with PE (mild and severe) compared with healthy women (5.71±0.414 ng/ml, 5.31±0.38 ng/ml and 6.01±0.282 ng/ml), respectively. Conclusions: Urinary PP13 and sFlt-1 levels in the third trimester of pregnancy were significantly correlated with the severity of PE, and urinary levels of PP13 were found to be decreased significantly in patients with PE than in healthy pregnant women in the 3rd trimester of pregnancy.

Circulating Vascular Endothelial Growth Factor and Soluble fms-Like Tyrosine Kinase-1 as Biomarkers for Endometrial Remodeling Across the Menstrual Cycle.

To characterize variation in circulating vascular endothelial growth factor (VEGF) and its receptor, soluble fms-like tyrosine kinase-1 (sFLT-1), across the menstrual cycle in normal ovulating women in relation to reproductive hormones to identify the utility of VEGF and sFLT-1 as peripheral biomarkers of endometrial remodeling. Ninety-six healthy, regularly menstruating ovulatory women, aged 18-44 years, enrolled in the BioCycle Study, a prospective cohort study at a U.S. academic research center. Vascular endothelial growth factor and sFLT-1 were measured in concurrently collected plasma, serum, and urine up to eight times across a single cycle. Reproductive hormones were measured in serum. Mean concentrations of VEGF and sFLT-1 were compared across phases of the cycle, and correlations between specimen types were calculated. Harmonic models estimated associations between VEGF and sFLT-1 and characteristics of hormonal patterns. No variation in VEGF or sFLT-1 levels were detected over the menstrual cycle. Median (25th percentile, 75th percentile) concentrations of VEGF during the menstrual cycle were 31.2 pg/mL (24.1, 56.9) in plasma, 194.1 pg/mL (125.4, 350.2) in serum, and 101.7 pg/mL (64.2, 165.8) in urine. Plasma and serum measures were consistently correlated, whereas urinary measures were not. Vascular endothelial growth factor was not consistently associated with reproductive hormone concentrations, although sFLT-1 was associated with higher mean and amplitude of estradiol. Circulating VEGF and sFLT-1 did not vary across the menstrual cycle and therefore are unlikely to be useful peripheral biomarkers of endometrial changes across the menstrual cycle. For studies measuring circulating VEGF for other reasons, plasma may be the preferred medium and timing to menstrual cycle phase need not be considered for reproductive-age women.

Modelling preeclampsia: a comparative analysis of the common human trophoblast cell lines.

ABSTRACTPreeclampsia remains a challenge without an effective therapy. Evidence supports targetability of soluble fms‐like tyrosine kinase‐1 (sFlt‐1) and soluble endoglin (sEng), which are released excessively from the placenta under ischemic and hypoxic stresses. We compared four trophoblast cell lines, BeWo, Jar, Jeg‐3, and HTR‐8/SVneo, in order to identify a suitable model for drug screening. Cultured trophoblasts were exposed to 1% oxygen vs. normoxia for 24‐48 hr; human umbilical vein and aortic endothelial cells were included for comparison. Supernatant sFlt‐1 and sEng concentrations were measured by ELISA, and sFlt‐1 mRNA expression determined by RT‐PCR. Cellular responses to experimental therapeutics were explored. All four trophoblast lines secreted sEng, which did not increase by hypoxia. BeWo, Jar, and Jeg‐3 exhibited significantly enhanced expression of sFlt‐1 i13 and e15a mRNA in response to hypoxia; however, only BeWo released a detectable level of sFlt‐1 protein, which was doubled by hypoxia. In contrast, hypoxia decreased sFlt‐1 mRNA expression and protein release in HTR‐8/SVneo, similarly to endothelial cells. The cellular mechanism involved HIFα. BeWo responded to representative agents similarly to human primary placental tissues in the literature. These data support that the BeWo‐hypoxia model mimics a key pathogenic mechanism of preeclampsia and has potential value for translational drug discovery.

Evaluation of Angiogenic Factors (PlGF and sFlt-1) in Pre-eclampsia Diagnosis.

Recent observations support the hypothesis that an imbalance between angiogenic factors has a fundamental role in the pathogenesis of pre-eclampsia and is responsible for the clinical manifestations of the disease. The goal of the present study was to evaluate the sensitivity, specificity, and the best accuracy level of Soluble fms-like tyrosine kinase-1 (sFlt-1), placental growth factor (PlGF), and sFlt-1/PlGF ratio in maternal serum and protein/creatinine ratio in urine sample to define the best cutoff point of these tests to discriminate between the patients with gestational hypertension and the patients with pre-eclampsia, to evaluate the possibility of using them as diagnostic methods. A prospective longitudinal study was performed, and blood samples were collected from 95 pregnant patients with hypertension to measure serum concentrations of biomarkers sFlt-1 and PlGF. Urine samples were collected for protein screening. Significance was set as p < 0.05. The sFlt-1/PlGF ratio demonstrated a sensitivity of 57.5% and a specificity of 60% using 50.4 as a cutoff point. The test that showed the best accuracy in the diagnosis of pre-eclampsia was protein/creatinine ratio, with a sensitivity of 78.9% and a specificity of 70% using 0.4 as a cutoff point and showing an area under the receiver operating characteristic curve of 0.80 (p < 0.001). No studied laboratory test proved to be fairly accurate for the diagnosis of pre-eclampsia, except for the protein/creatinine ratio. The evidence is insufficient to recommend biomarkers sFlt-1 and PlGF to be used for the diagnosis of pre-eclampsia.

Prediction of adverse maternal outcomes of early severe preeclampsia.

Assess soluble FMS-like tyrosine kinase-1/placental growth factor ratio (sFlt-1/PlGF) diagnostic accuracy for predicting adverse maternal outcome in patients with early severe preeclampsia, and whether its predictive performance is superior to full preeclampsia integrated estimate of risk score (PIERS). Prospective study enrolled patients with early severe preeclampsia (defined by American College of Obstetricians and Gynaecologists 2013 guidelines) admitted to the Clinic for Obstetrics and Gynaecology, Clinical Center of Serbia intensive care unit. Patients underwent delivery to terminate preeclampsia within 48h of admission. PIERS was generated and blood samples taken at admission. Multiple pregnancies and gestational ages outside 24-34weeks were excluded. sFlt-1 and PlGF serum concentrations were measured using Elecsys® assays and cobas e 601 analyser. Maternal complications were recorded for seven days post-delivery. Diagnostic accuracy (sensitivity and specificity), and predictive performance (receiver operating characteristic area under curve [AUC]) vs. PIERS, of sFlt-1/PlGF for predicting adverse maternal outcome. Of 89 patients enrolled, 61 were evaluable. Median frequency of adverse maternal outcomes within seven days of delivery was two. Median sFlt-1/PlGF and PIERS were 521·0 and 5·0%, respectively. sFlt-1/PlGF showed greater correlation with complication number than PIERS (Spearman's rho: 0·728 [p<0·001] and 0·134 [p=0·304], respectively). AUC for sFlt-1/PlGF and PIERS were 0·853 and 0·628, respectively. A 377·0 sFlt-1/PlGF cut-off was optimal for predicting complications (75·0% sensitivity; 92·3% specificity). sFlt-1/PlGF correlated more closely with number of adverse maternal outcomes than PIERS, and was a superior predictor of maternal complications.

PLGF/ SFLT-1 Concentrations As Early Predictors Preeclampsia In Pregnancy During Pandemic Covid-19

PLGF/ SFLT-1 Concentrations As Early Predictors Preeclampsia In Pregnancy During Pandemic Covid-19

The soluble VEGF receptor sFlt-1 contributes to endothelial dysfunction in IgA nephropathy.

Endothelial injury is a common manifestation in IgA nephropathy (IgAN). After the previous identification of the upregulated soluble fms-like tyrosine kinase-1 (sFlt-1) correlated with endothelial injury in IgAN, in the present study, we further explored the role of sFlt-1 in endothelial injury in IgAN. We enrolled 72 patients with IgAN and detected the sFlt-1 levels. The polymeric IgA1 (pIgA1) complexes were isolated from the pooled plasma samples of another 10 patients with IgAN. Apoptosis proteins were detected in cultured human umbilical vein endothelial cells (HUVECs) with the stimulation of recombinant sFlt-1 or the caspase-9 inhibitor Z-LEHD-FMK. We identified there were positive correlations between sFlt-1 and IgA-IgG complex as well as vWF levels in patients with IgAN. The sFlt-1 levels in HUVECs were significantly upregulated by pIgA1 complex derived from IgAN patients in a concentration-dependent manner. The proliferation ability of HUVECs was damaged when stimulated with sFlt-1 protein in a time- and dose- dependent manner. And the apoptosis rate was up-regulated significantly as the stimulation concentrations of sFlt-1 increased. We found sFlt-1 challenge could significantly increase the expression of vWF. In addition, sFlt-1 increased the levels of caspase-9, caspase-3, Bax and mitochondrial membrane potential; facilitated the release of cytochrome C from mitochondria to cytoplasma. In contrast, Z-LEHD-FMK attenuated high sFlt-1-induced HUVECs apoptosis. In conclusion, our study demonstrated that sFlt-1 expression was up-regulated by the challenge of pIgA1 complex derived from patients with IgAN. Furthermore, increased sFlt-1 facilitated human umbilical vein endothelial cells apoptosis via the mitochondrial-dependent pathway.

Quercetin attenuates reduced uterine perfusion pressure -induced hypertension in pregnant rats through regulation of endothelin-1 and endothelin-1 type A receptor

BackgroundQuercetin was reported to be crucial for a broad range of activities, including attenuating inflammation, platelet aggregation, capillary permeability, and lipid peroxidation. However, the effect of quercetin in hypertension during pregnancy, was not fully understood.MethodsThe model of hypertension in pregnancy was established in rats by reduced uterine perfusion pressure (RUPP). Quercetin was administrated by gavage. Systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured using the CODA 6 BP system. Plasma concentrations of Endothelin-1 (ET-1), soluble fms-like tyrosine kinase-1 (sFlt-1), and vascular endothelial growth factor (VEGF) were detected using enzyme-linked immunosorbent assay kits. The mRNA and protein levels of ET-1 and endothelin-1 type A receptor (ETAR) were determined by RT-PCR and Western blotting. The ETAR antagonist BQ-123 was performed by osmotic minipumps.ResultsIn RUPP induced rats, quercetin treatment decreased SBP and DBP, fetal resorptions percentage, plasma ET-1 and sFlt-1 concentrations, ET-1 and ETAR levels, but increased fetal body weight and VEGF expression. BQ-123 administration attenuated SBP and DBP, suppressed fatal resorptions percentage, and increased fetal body weight of RUPP rats.ConclusionQuercetin attenuates RUPP induced hypertension in pregnant rats through the regulation of ET-1 and ETAR.

Placental Growth Factor and Soluble, Fms-Like Tyrosine Kinase-1 in Preeclampsia: A Case-Cohort (PEARL) Study.

Preeclampsia is associated with a higher maternal blood levels of soluble fms-like tyrosine kinase-1 (sFlt-1) and lower levels of placental growth factor (PlGF) that appear before clinical onset. We aimed to estimate the normal progression of these biomarkers in normal pregnancies and in those affected by preeclampsia. We conducted a case-cohort study including low-risk nulliparous women recruited at 11-13 weeks gestation (cohort) and women with preeclampsia (cases). Maternal blood was collected at different points during pregnancy including at the time of diagnosis of preeclampsia for cases. Maternal serum PlGF and sFlt-1 concentrations and the sFlt-1/PlGF ratio were measured using B•R•A•H•M•S plus KRYPTOR automated assays and were compared between patients in both groups matched for gestational age. Cases were stratified as early- (≤34 weeks), intermediate- (35-37 weeks) and late-onset (>37 weeks) preeclampsia. The cohort consisted of 45 women whose results were compared with those of 31 women who developed preeclampsia, diagnosed at a median gestational age of 32 weeks (range 25-38 weeks). We observed that sFlt-1, PlGF and their ratio fluctuated during pregnancy in both groups, with a significant correlation with gestational age after 28 weeks (P < 0.05). We observed a significant difference between cases and controls, with a median ratio 100 times higher in early preeclampsia (P < 0.001), 13 times higher in intermediate preeclampsia (P < 0.001), but no significant difference between groups in late-onset preeclampsia with matched controls. PlGF, sFlt-1, and their ratio may be useful in the prediction and diagnosis of early- and intermediate-onset preeclampsia but are not useful for late-onset preeclampsia.

The sFlt-1/PlGF ratio as a triage tool to identify superimposed preeclampsia in women with chronic hypertension in emergency rooms.

Assess the usefulness of the sFlt-1/PlGF ratio for the differential diagnosis of uncontrolled chronic hypertension vs. superimposed preeclampsia. We performed a cross-sectional study from 2015 to 2017 and 42 women with initial diagnosis of superimposed preeclampsia were enrolled in the emergency room. After a 12week follow up patients were grouped as superimposed preeclampsia (Group A) and uncontrolled chronic hypertension (Group B) according to the American College of Obstetricians and Gynecologist criteria. A group of 33 healthy women paired by gestational age were included as controls (Group C). Maternal serum levels of sFlt-1 and PlGF were measured at enrollment, and the ratios of the groups were compared. Superimposed preeclampsia vs. uncontrolled chronic hypertension. After follow-up, group distribution was 30 women in Group A, 12 women in Group B, and 25 women in Group C. The sFlt-1/PlGF ratio was higher in women with superimposed preeclampsia than in women with uncontrolled chronic hypertension (215.5 vs. 9.65, p<0.001). The control group displayed lower ratio values (3.66, p<0.001). The sFlt-1 concentration was higher in Group A than in Group B (7564 vs. 1281pg/mL, p<0.001) and the PlGF level was lower in Group A (34.39 vs. 169pg/mL, p<0.001). The sFlt-1/PlGF ratio exhibits good performance for the differential diagnosis of superimposed preeclampsia vs. uncontrolled chronic hypertension.

Enhanced Serum Levels of sFlt1: Impact on Materno–Fetal CMV Transmission

Background: Antenatal Cytomegalovirus infection (CMV) can be associated with severe fetal symptoms and newborn outcome. The current prenatal diagnosis is based on amniocentesis (AC). No reliable biomarker for fetal infection is available. Methods: We measured Placenta-derived growth factor (PlGF), and soluble fms-like tyrosine kinase 1 (sFlt1), concentrations in maternal serum and amniotic fluid (AF) in context of maternal CMV primary infection. Blood sampling was carried out at the time of AC for detection of fetal CMV infection. The study cohort was divided into four subcohorts according to the presence or absence of fetal infection and preemptive hyperimmunoglobulin (HIG) treatment during the time interval between diagnosis of the CMV primary infection and AC. Results: The study cohort involved 114 pregnancies. In the non-transmitting subcohorts (NT) with and without prior HIG treatment, the median sFlt1 concentrations were 1.5 ng/mL (NT, HIG+) and 1.4 ng/mL (NT, HIG−), respectively. In the two transmitting groups (T) the concentrations were 1.3 ng/mL (T, HIG+) and 2.3 ng/mL (T, HIG−), respectively (NT, HIG− vs. T, HIG−, p < 0.001). The corresponding PlGF levels and the sFlt1/PlGF ratios showed no significant differences between the cohorts. The empirical cut-off values <1504 pg/mL sFlt1 and <307 pg/mL PlGF, were associated with the exclusion of CMV transmission (p < 0.001). Conclusion: sFlt1 concentration in the maternal blood could be a predictive biomarker for maternofetal CMV transmission.

Calcium deficient placental growth restriction is mediated by an increase in non-invasive integrin α5 and β4 phenotype.

Integrins are cell adhesion receptors that participated in endovascular invasion by cytotrophoblasts in preeclampsia. This study aimed to investigate the effect of calcium on cellular pathways influencing the trophoblast integration into endothelial cellular networks in vitro. Red fluorescent-labelled human uterine myometrial microvascular endothelial cells (UtMVECs) were seeded on Matrigel. Green fluorescent-labelled HTR-8/SVneo trophoblast cells were then co-cultured with endothelial cells in different concentrations of calcium for 24h. The calcium effects on HTR-8/SVneo cell integration were quantified by Image J. Quantitative PCR was performed to measure mRNA expression of integrins α1, α5, α6, β1 and β4. The concentrations of interleukin IL-6, matrix metalloproteinase-2 (MMP-2), MMP-9, PlGF and sFlt-1 in the conditioned medium were measured by ELISA while levels of cytokines IL-1β, IL-8, IL-10, TNF-α and INF-γ were assessed by magnetic Luminex assays™. Both calcium depletion (0.4mM) and low calcium (1.8mM) groups demonstrated inhibited integration of trophoblast cells into endothelial cellular networks, compared with the normal calcium group (2.4mM). The IL-6 production was reduced from conditioned media in both calcium depletion and low calcium groups. In calcium depletion group, mRNA expression of integrin α5 and β4 in trophoblasts was increased while integrin α1 was decreased. The in vitro trophoblast cell integration into endothelial cellular networks could be modified by altering media calcium through integrin switch away from integrins α5 and β4 and towards integrin α1 which may be required for healthy early trophoblast integration.