sFlt-1 Levels Research Articles

Angiogenic factors for prediction of preeclampsia and intrauterine growth restriction onset in high-risk women: AngioPred study

Objective: The study aimed to compare the level of two angiogenic factors, soluble fms-like tyrosine kinase-1 (sFlt1) and soluble endoglin (sEng), for the prediction of preeclampsia and intrauterine growth restriction in high-risk pregnant women.Methods: A prospective multicenter cohort study of 200 pregnant patients was conducted between June 2008 and October 2010. sFlt1 and sEng were measured by enzyme-linked immunosorbent assay.Results: Forty-five patients developed a placenta-mediated adverse pregnancy outcome. Plasma levels of sFlt1 and sEng were higher in patients who will experience a preeclampsia at 28, 32, and 36 weeks compared with patients with no complication. The same results were observed for intrauterine growth restriction. Plasma levels of sFlt1 and sEng were not significantly different for patients with preeclampsia compare to patients with intrauterine growth restriction. Patients with early pre-eclampsia (PE) had very high rates of angiogenic factors at 20, 24, and 28 weeks. Patients with late PE and early and late intrauterine growth retardation (IUGR) had high rates at 32 and 36 weeks.Conclusion: In high-risk women, angiogenic factors are disturbed before the onset of preeclampsia and this is true for intrauterine growth restriction.

Tobacco exposure and wound healing in head and neck surgical wounds.

Smoking impairs wound healing, yet the underlying pathophysiological mechanisms are unclear. We evaluated tobacco-altered healing in head and neck surgery by studying the association between biomarkers and tobacco exposure, as well as cutaneous perfusion by smoking status. Prospective cohort study, tertiary/academic care center, 2011 to present. Patients who required head and neck surgery were enrolled prospectively. Postsurgical drain fluid was collected 24 hours postoperatively. Biomarkers associated with postulated mechanisms of smoking-impaired healing were assayed. These included interleukin-1, -6, and -8; tumor necrosis factor- alpha; transforming growth factor-beta; epidermal growth factor (EGF); basic fibroblastic growth factor (bFGF); C-reactive protein; vascular endothelial growth factor; soluble FMS-like tyrosine kinase-1 (sFLT-1); and placental growth factor. Tobacco exposure and clinical outcomes were recorded. Two sample two-sided t tests evaluated the differences in cytokine levels by tobacco exposure. In a second cohort, cutaneous vascular assessment via indocyanine green angiography was compared by smoking status. Twenty-eight patients were enrolled with drain fluid collection. Twenty-one subjects were current/former smokers, whereas seven were never smokers. EGF was higher in never smokers than smokers in a statistically significant manner (P = 0.030). Likewise, sFLT-1 was significantly higher in never smokers (P = 0.011). Cutaneous angiography revealed nonsmokers to have significantly higher cutaneous perfusion than smokers. In this head and neck surgical cohort, significantly higher EGF and sFLT-1 levels in wound fluid were associated with never smoking, suggesting that smoking has adverse effects on the inflammatory phase of wound healing. Cutaneous angiography supports the detrimental effect of smoking on skin perfusion. These findings suggest the need for further study as well as therapeutic targets for smokers undergoing surgery. 2b. Laryngoscope, 128:618-625, 2018.

Abstract 040: Low Soluble Fms-like Tyrosine Kinase-1, Endoglin, and Endothelin-1 Levels in Women With Confirmed or Suspected Preeclampsia Using Proton Pump Inhibitors

Preeclamptic patients display elevated placenta-derived soluble Fms-like tyrosine kinase-1 (sFlt-1) and endoglin levels, and decreased placental growth factor (PlGF) levels. Proton pump inhibitors (PPIs) decrease trophoblast sFlt-1 and endoglin secretion in vitro. PPIs are used during pregnancy, to combat reflux disease. Here we investigated whether PPIs affect sFlt-1 in women with confirmed/suspected preeclampsia, making use of a prospective cohort study involving 430 women. Of these women, 40 took PPIs (6 esomeprazole, 32 omeprazole and 2 pantoprazole) for 8-45 (median 29) days before sFlt-1 measurement. Measurements were only made once, at study entry between weeks 20-41 (median 33 weeks). PPI use associated with lower sFlt-1 levels, with no change in PlGF levels, both when compared to all non-PPI users, and to 80 gestational age-matched controls selected from the non-PPI users. No sFlt-1/PlGF alterations were observed in women using ferrous fumarate or macrogol, while, as expected, women using antihypertensive medication displayed higher sFlt-1 levels and/or lower PlGF levels. The PPI use-associated decrease in sFlt-1 was independent of the application of antihypertensive drugs and also occurred when restricting our analysis to patients with hypertensive disease of pregnancy at study entry. PPI users displayed more cases with preexisting proteinuria, less gestational hypertension, and a lower number of neonatal sepsis cases. Finally, their plasma endoglin and endothelin-1 levels were lower, while sFlt-1 levels correlated positively with both. In conclusion, PPI use associates with low sFlt-1, endoglin, and endothelin-1 levels, warranting prospective trials to investigate the therapeutic potential of PPIs in preeclampsia.

Abstract P301: Preeclampsia and Gestational Hypertension: Does sEng Levels Make the Important Difference Preeclampsia and Gestational Hypertension: Does sEng Levels Make the Important Difference

The origin of Preeclampsia (PE), a multi-systemic disorder of pregnancy, lies to a great extent on the balance of the anti-angiogenic and angiogenic proteins during pregnancy. In the present study, we have determined the levels of sFlt-1 and sEng levels (anti-angiogenic protein) and Placental Growth Factor, PlGF (angiogenic Protein) in the serum of the patients with hypertensive disorder of pregnancy (including Gestational Hypertension, GH and PE) and Normotensive (NT) pregnancy. The levels of sFlt-1 were found to be significantly higher in the 3 rd trimester and at term in the hypertensive pregnancy (GH and PE), compared to the NT pregnancy, (8.67±1.93 ng/ml, n=13 vs. 3.95±.37 ng/ml, n=111 at the 3rd trimester and 14.90±2.68 ng/ml, n=15 vs. 6.58±.42 ng/ml, n=111 at term). The levels of sFlt-1 were considerably higher in the PE samples compared to the GH samples, but not significant. Similarly the levels of sEng were found to be significantly higher in the 3 rd trimester and at term in the hypertensive pregnancy (GH and PE), compared to the NT pregnancy (11.18±1.51 ng/ml, n=15 vs. 7.86±.25 ng/ml, n=106 at the 3 rd trimester and 12.68±1.70 ng/ml, n=14 vs. 10.01±.42 ng/ml, n=106 at the term). The levels of sEng were, however, found to be significantly higher in all the trimester and at term in the PE samples compared to the NT Samples ( 6.49±1.14 ng/ml, n=7 vs 4.21±.28 ng/ml, n=106, 6.89±2.30 ng/ml, n=7 vs. 3.76±.25 ng/ml, n=106, 13.10±2.12 ng/ml, n=7 vs. 7.86±.37 ng/ml, n=106, 14.93±2.27 ng/ml, n=7, vs. 10.01±.42 ng/ml, n=106 at the 1 st , 2 nd , 3 rd trimesters and term respectively), but not in the GH samples. The angiogenic protein, PlGF was found to be significantly lower in the hypertensive conditions of pregnancy (PE and GH) samples compared to the NT samples, only in the 3 rd trimester (440±48.58 pg/ml, n=15 vs 1278±104 pg/ml, n=119), although its levels were considerably decreased in the 2 nd trimester and at term, but not significant. Within the hypertensive conditions of pregnancy, the blood pressure increase at term was more in the PE patients compared to the GH patients (42±5.1/25±3.3 mm Hg vs. 18±3.7/13±3.3 mm Hg, p<0.05 ) compared to the BP recorded at the 1 st trimester. The results of the present study indicate a more potent role of sEng in the development of PE.

Soluble Fms-Like Tyrosine Kinase 1 (sFlt-1) and Risk of Cerebral Vasospasm After Aneurysmal Subarachnoid Hemorrhage

The molecular mechanisms underlying cerebral vasospasm and delayed cerebral ischemia (DCI) after aneurysmal subarachnoid hemorrhage (aSAH) are incompletely understood. We hypothesized that circulating antiangiogenic factors, such as soluble Fms-like tyrosine kinase 1 (sFlt-1) and soluble transforming growth factor β coreceptor, soluble endoglin (sEng), are important markers of their pathophysiology. We performed a prospective study in patients with aSAH and measured cerebrospinal fluid and serum levels of sFlt-1 and sEng on postbleed day 1 and 6 and correlated levels with incidence and severity of cerebral vasospasm and DCI. Twenty-seven patients with aSAH were enrolled in the study. Severe angiographic vasospasm was present in 14.8% of patients and DCI occurred in 33.3%. Serum sFlt1 levels were increased on postbleed day 6 in patients who developed vasospasm. However, on postbleed day 1, there were no differences in patients who developed vasospasm. Increased serum sFlt-1 levels on postbleed day 1 were found to predict the development of severe angiographic vasospasm with an area under the curve of 0.818 with an optimal cutoff value of 95 pg/mL. Alterations in sFlt1 were not associated with DCI. Serum and cerebrospinal fluid sEng levels did not correlate with vasospasm or DCI. Serum levels of sFlt-1 are increased in patients with aSAH who are at risk for severe vasospasm. Further studies with larger sample sizes are needed to evaluate whether sFlt-1 levels may predict onset of severe vasospasm and DCI.

Serum levels of PTX3 and SFLT-1 predict the outcomes of frozen-thawed euploid embryo transfer cycles

Serum levels of PTX3 and SFLT-1 predict the outcomes of frozen-thawed euploid embryo transfer cycles

Abstract 037: Exaggerated Placental Ischemia-induced Hypertension in Endothelin Receptor Type B (ETB)-deficient Pregnant Rats s Independent of Increased sFlt-1 or ROS Levels

While the pathogenesis of preeclampsia is not fully understood, studies implicate placental ischemia. Reduced uterine perfusion pressure (RUPP)-induced placental ischemia/hypoxia in animal models stimulates release of factors like antiangiogenic sFlt-1 into the maternal circulation increasing vascular-renal ET-1. ET-1 promotes hypertension via reactive oxygen species (ROS). Blockade of vasoconstrictive ETA abolishes RUPP hypertension. Deficiency of vasodilatory ETB in rats leads to increased blood pressure in pregnancy. While ETB deficiency markedly enhances RUPP hypertension, it is unknown if there is exaggerated RUPP-induced sFlt-1, ET-1 or ROS levels in ETB-def rats. The hypothesis was tested that placental ischemia/hypoxia-induced release of sFlt-1 and circulating ET-1 and ROS are greater in ETB-def rats. Eighteen-week-old ETB-def and transgenic (Tg) control pregnant rats were generated with Wistar Hannover males. RUPP or Sham surgeries were on gestational day 14 and assessment of plasmas and placentas at day 19. RUPP increased placental sFlt-1 (pg/mg) similarly in RUPP ETB-def (781±113, N=5) vs Sham ETB-def (573±54, N=12) and RUPP Tg (631±62, N=5) vs Sham Tg (547±31, N=12) (P<0.05). In placental explant cultures, acute hypoxia (48 h 1% O2 vs normoxia 6% O2) stimulated a comparable release of sFlt-1 (pg/mg) in Sham ETB-def (2577±135 vs 2070±78) and Sham Tg (3208±318 vs 2553±107) (P<0.05). Unexpectedly, plasma sFlt-1 (pg/mL) was lower in RUPP ETB-def (153±48) vs Sham ETB-def (476±125) and RUPP Tg (238±32) vs Sham Tg (463±102) (P<0.05). Plasma ET-1 (fmol/L) was exaggerated in RUPP ETB-def (954±70) and greater in Sham ETB-def (735±43) vs RUPP Tg (122±14) or Sham Tg (142±41) (P<0.05). Plasma H2O2 (umol/L) was not exaggerated in RUPP ETB-def (5.4±1.2) or RUPP Tg (4.0±0.5) but was greater (P<0.05) in Sham ETB-def (6.2±0.3) vs Sham Tg (3.6±0.3). In conclusion, these data suggest in 1) normal pregnancy, ETB is crucial for blood pressure control by regulating bioavailable ET-1 to prevent ROS production and 2) placental ischemia, ETB reduces excess ET-1 to buffer hypertension independently of sFlt-1 or ROS. These data support ETB physiology as important in controlling blood pressure in pregnancy and its loss in mediating hypertension in preeclampsia.

Abstract 038: MiR-210 Overexpression Induces sFlt1 Production and Contributes to Preeclampsia-like Symptoms in Pregnant Mice

Preeclampsia (PE) is an obstetric complication that is diagnosed by hypertension and proteinuria at or after 20 weeks of gestation. PE is a multifactorial disease, but it is widely accepted that impaired proangiogenic factors such as vascular endothelial growth factor (VEGF), placental growth factor (PlGF) and antiangiogenic factor such as soluble FMS-like tyrosine kinase-1 (sFlt1) or VEGF receptor-1 balance during pregnancy is linked to PE. sFlt1, a splice variant of Flt1 binds to either VEGF or PlGF and blocks their action. In addition, placental angiogenesis is known to be regulated by specific miRNAs that are typically dysregulated during PE highlighting the importance of these miRNAs in pregnancy disorders. miR-210 is upregulated in placentas and serum/plasma of PE women and is also known to modulate angiogenic pathways. Therefore, we hypothesized that miR-210 overexpression contributes to the development of PE-like symptoms by altering the angiogenic/antiangiogenic pathway. To this end, we demonstrate increased systolic blood pressure (SBP: miR-210 TG P = 115 ± 2 mm Hg vs. WT P = 95 ± 2 mm Hg, p<0.05), endothelial dysfunction and proteinuria in pregnant miR-210 TG mice as compared to control WT mice. Placental sFlt-1 levels increased and PlGF levels decreased in miR-210 TG mice as determined by immunoblotting. Immunohistochemistry (IHC) analysis demonstrated an increase in sFlt1 and a decrease in PlGF immunoreactivity in miR-210 TG placentas. Pregnant miR-210 TG mice exhibited increased serum levels of sFlt-1 as well as decreased PlGF as compared to pregnant WT mice. Furthermore, immunohistological analysis of stained human placental sections revealed an increase in sFlt-1 and a decrease in PlGF in pregnant miR-210 TG as compared to pregnant WT mice. To determine the placental etiology, human cytotrophoblasts (CTBs) overexpressing miR-210 demonstrated a decrease in PlGF while inhibition of miR-210 in CTBs using anti-miR-210 oligos increased the PlGF expression as determined by immunoblotting. These data taken together suggest that miR-210 modulates the angiogenic/antiangiogenic pathway and contributes to PE-like features in mice. Thus, targeting miR-210 by delivering anti-miR-210 oligos may prevent PE-like symptoms in pregnant women.

Ferulic acid alleviates symptoms of preeclampsia in rats by upregulating vascular endothelial growth factor.

Preeclampsia is a complication affecting pregnant women worldwide, which leads to maternal and fetal morbidity and mortality. In this study, we evaluated the efficacy of ferulic acid (FA) on an Nω -nitro-L-arginine methyl ester hydrochloride (L-NAME) induced rat model of preeclampsia. L-NAME was administered to pregnant rats to induce preeclampsia. 48 rats were divided into three experimental groups (n=16 each): control group, preeclampsia group and preeclampsia with FA treatment (preeclampsia+FA). Physiological characteristics such as urine volume, total urine protein and blood pressure were assessed. Expressions levels of urinary nephrin and podocin mRNAs were analyzed by RT-PCR. Levels of renal vascular endothelial growth factor (VEGF), renal soluble fms-like tyrosine kinase-1 (sFlt-1) and serum placenta growth factor (PlGF) were also examined. Urine volume, total urine protein and blood pressure were markedly increased in preeclampsia group rats compared to control (P<.05), which were then significantly reduced in preeclampsia+FA group (P<.05). Expressions of urinary nephrin and podocin mRNAs, levels of VEGF, sFlt-1 and PlGF were also reversed in preeclampsia+FA group compared to preeclampsia rats (P<.05). We hereby report for the first time, FA alleviates preeclampsia symptoms in a rat preeclampsia model, supporting its potential value in treating preeclampsia.

Modulating circulating sFlt1 in an animal model of preeclampsia using PAMAM nanoparticles for siRNA delivery

IntroductionExcessive circulating sFlt1 plays a major role in the pathogenesis of preeclampsia (PE). Using RNAi to silence sFlt1 may be a therapy for treating PE. Because of the rapid degradation of siRNA, gene therapy in vivo remains limited. Poly-amidoamine (PAMAM) has been demonstrated to be an excellent nanocarrier for siRNA delivery with no discernible toxicity. MethodsThe aim of the present study was to investigate the therapeutic effect of siRNA-sFlt1-PAMAM on PE. The biophysical properties of siRNA-sFlt1-PAMAM complexes were analysed. Next, HTR-8/SVneo cells were incubated with the complexes. The transfection efficiency, silencing effect, and cell proliferation were examined. The timed pregnant rats were grouped to test the in vivo effect of siRNA-sFlt1-PAMAM on PE. The PE model was established by injection of tumour necrosis factor-α (TNF-α), while treatment group rats were injected with both TNF-α and siRNA-sFlt1-PAMAM. The mean arterial pressure, serum levels of sFlt1, and 24-h urinary protein were measured. The number of viable pups, and weights of individual pups and placentae were determined. ResultssiRNA-sFlt1-PAMAM complexes showed high cellular uptake and excellent siRNA encapsulation ability. siRNA-sFlt1-PAMAM significantly decreased sFlt1 secretion from HTR-8/SVneo cells (p = 0.001) with little effect on HTR-8/SVneo cell proliferation. The circulating level of sFlt1, mean arterial pressure, and urine protein level in the TNF-α group were significantly increased compared to that in the control group, while the weights of foetuses and placentae were significantly decreased compared to that in the control group (p < 0.05). All preeclamptic symptoms were greatly attenuated by siRNA-sFlt1-PAMAM (p < 0.05). ConclusionsThese data suggest that siRNA-sFlt1-PAMAM effectively decreased sFlt1 secretion and improved pregnancy outcomes in a preeclamptic rat model, which may provide a new therapeutic strategy for PE.

Potential New Non-Invasive Therapy Using Artificial Oxygen Carriers for Pre-Eclampsia.

The molecular mechanisms of pre-eclampsia are being increasingly clarified in animals and humans. With the uncovering of these mechanisms, preventive therapy strategies using chronic infusion of adrenomedullin, vascular endothelial growth factor-121 (VEGF-121), losartan, and sildenafil have been proposed to block narrow spiral artery formation in the placenta by suppressing related possible factors for pre-eclampsia. However, although such preventive treatments have been partly successful, they have failed in ameliorating fetal growth restriction and carry the risk of possible side-effects of drugs on pregnant mothers. In this study, we attempted to develop a new symptomatic treatment for pre-eclampsia by directly rescuing placental ischemia with artificial oxygen carriers (hemoglobin vesicles: HbV) since previous data indicate that placental ischemia/hypoxia may alone be sufficient to lead to pre-eclampsia through up-regulation of sFlt-1, one of the main candidate molecules for the cause of pre-eclampsia. Using a rat model, the present study demonstrated that a simple treatment using hemoglobin vesicles for placental ischemia rescues placental and fetal hypoxia, leading to appropriate fetal growth. The present study is the first to demonstrate hemoglobin vesicles successfully decreasing maternal plasma levels of sFlt-1 and ameliorating fetal growth restriction in the pre-eclampsia rat model (p < 0.05, one-way ANOVA). In future, chronic infusion of hemoglobin vesicles could be a potential effective and noninvasive therapy for delaying or even alleviating the need for Caesarean sections in pre-eclampsia.

Micro- and Nano-vesicles from First Trimester Human Placentae Carry Flt-1 and Levels Are Increased in Severe Preeclampsia.

Preeclampsia is a life-threatening hypertensive disease affecting 3-5% of pregnancies. While the pathogenesis of preeclampsia remains unclear, it is known that placenta-derived factors trigger the disease by activating maternal endothelial cells prior to the onset of clinical symptoms. Extracellular vesicles (EVs) of different sizes extruded by the placenta may be one factor. The truncated/secreted form of Flt-1 (sFlt-1) has also been implicated in the pathogenesis of preeclampsia. We investigated whether placental EV production is altered in preeclampsia such that they induce endothelial cell activation, and whether (s)Flt-1 is involved. Macro-, micro-, and nano-vesicles were collected from normal and preeclamptic (PE) placental explants, and separated by differential centrifugation. The number and size of micro- and nano-vesicles was measured by nanoparticle tracking analysis and their ability to activate endothelial cells was quantified by endothelial cell intercellular adhesion molecule 1 expression and monocyte adhesion. The levels of Flt-1 were measured by western blots and ELISA. PE placentae extruded significantly more micro- and nano-vesicles than control placentae and the extruded micro-vesicles were larger than those from control placentae. Micro- and nano-vesicles from both first trimester and term human placentae carried Flt-1 and levels were significantly increased in EVs from severe, but not mild, PE compared to normotensive placentae. All fractions of EVs from PE placentae activated endothelial cells, and for micro- and nano-vesicles, activation was reduced in the presence of exogenous vascular endothelial growth factor (VEGF), a Flt-1 neutralizing antibody, or by pre-treatment with VEGF. While EV-bound VEGF constituted over 20% of the total detected VEGF secreted by PE and normotensive placentae, EV-bound Flt-1 did not significantly contribute to the total level of sFlt-1/Flt-1 released by human third trimester placentae. Micro- and nano-vesicles extruded by human placentae carry Flt-1 across gestation and in severe preeclampsia, the levels of vesicle-bound Flt-1 are upregulated. All fractions of PE placental EVs activated endothelial cells and for micro- and nano-vesicles, this was in part due to the ability of EV-bound Flt-1 to sequester VEGF. That placental EVs can activate endothelial cells supports the contention that EVs are one placental toxin contributing to the pathogenesis of preeclampsia.

Low Soluble Fms-Like Tyrosine Kinase-1, Endoglin, and Endothelin-1 Levels in Women With Confirmed or Suspected Preeclampsia Using Proton Pump Inhibitors.

Patients with preeclampsia display elevated placenta-derived sFlt-1 (soluble Fms-like tyrosine kinase-1) and endoglin levels and decreased placental growth factor levels. Proton pump inhibitors (PPIs) decrease trophoblast sFlt-1 and endoglin secretion in vitro. PPIs are used during pregnancy to combat reflux disease. Here, we investigated whether PPIs affect sFlt-1 in women with confirmed/suspected preeclampsia, making use of a prospective cohort study involving 430 women. Of these women, 40 took PPIs (6 esomeprazole, 32 omeprazole, and 2 pantoprazole) for 8 to 45 (median 29) days before sFlt-1 measurement. Measurements were only made once, at study entry between weeks 20 and 41 (median 33 weeks). PPI use was associated with lower sFlt-1 levels, with no change in placental growth factor levels, both when compared with all non-PPI users and with 80 gestational age-matched controls selected from the non-PPI users. No sFlt-1/placental growth factor alterations were observed in women using ferrous fumarate or macrogol while, as expected, women using antihypertensive medication displayed higher sFlt-1 levels and lower placental growth factor levels. The PPI use-associated decrease in sFlt-1 was independent of the application of antihypertensive drugs and also occurred when restricting our analysis to patients with hypertensive disease of pregnancy at study entry. PPI users displayed more cases with preexisting proteinuria, less gestational hypertension, and a lower number of neonatal sepsis cases. Finally, their plasma endoglin and endothelin-1 levels were lower while sFlt-1 levels correlated positively with both. In conclusion, PPI use associates with low sFlt-1, endoglin, and endothelin-1 levels, warranting prospective trials to investigate the therapeutic potential of PPIs in preeclampsia.

P 48 Low soluble Fms-like tyrosine kinase-1, endothelin-1 and endoglin levels in women with suspected or confirmed preeclampsia, using proton pump inhibitors

Preeclamptic patients display elevated placenta-derived soluble Fms-like tyrosine kinase-1 (sFlt-1), endoglin and endothelin-1 levels and decreased placental growth factor (PlGF) levels. Proton pump inhibitors (PPIs) have been reported to decrease trophoblast sFlt-1 secretion in vitro. PPIs are widely used during pregnancy, to combat reflux disease. To investigate whether PPIs affect sFlt-1, endoglin, endothelin-1 and PlGF in women with (suspected) preeclampsia. This is a secondary analysis of a prospective cohort study involving 430 singleton pregnant women with suspicion or with confirmed preeclampsia from 20 weeks gestation. As a comparator of the PPIs, the effects of other drugs commonly used in women with suspected or confirmed preeclampsia, i.e., antihypertensive drugs (α-methyldopa, nifedipine), corticosteroids, ferrous fumarate and macrogol, is simultaneously assessed. 40 (9%) out of the 430 patients were treated with PPIs (6 esomeprazol, 32 omeprazol and 2 pantoprazol) for 8–45 (mean 29) days before blood sampling. PPI use was associated with lower sFlt-1 levels, with no change in PlGF levels, both when compared to all non-PPI users, and to 80 gestational age-matched controls selected from the non-PPI users. No sFlt-1 or PlGF alterations were observed in women using ferrous fumarate or macrogol, while, as expected, women using antihypertensive medication displayed higher sFlt-1 levels and/or lower PlGF levels. The PPI use-associated decrease in sFlt-1 was independent of the application of antihypertensive drugs and also occurred when restricting our analysis to patients with hypertensive disease of pregnancy at study entry. PPI users displayed more cases with preexisting proteinuria, less gestational hypertension, and a lower number of neonatal sepsis cases. Furthermore, their plasma endoglin levels were lower along with their plasma endothelin-1 levels, while sFlt-1 levels correlated positively with endothelin-1 levels. PPI use associates with low sFlt-1, endothelin-1 and endoglin levels, warranting prospective trials to investigate the therapeutic potential of PPIs in preeclampsia.

OP 14 Angiogenic factors in pregnancies complicated by rheumatoid arthritis

Introduction Women with active rheumatoid arthritis (RA) during pregnancy deliver often infants with reduced birth weight. We hypothesized that RA might have an impact on placental function and subsequently on the angiogenic profile. Objectives We aimed to investigate the pro- and anti-angiogenic factors such as placental growths factor (PLGF) and soluble fms-like tyrosine kinase-1 (sFlt-1), respectively, in patients with RA and healthy controls. Patients and methods PLGF and sFlt-1 serum levels of pregnant patients with RA ( n = 29) and gestational aged-matched healthy women ( n = 10) were analyzed once at each trimester (gestational week 10–12, 20–22 and 30–32). Disease activity of all RA patients was measured by DAS28-CRP and CRP. Neonatal birth weights and birth weight percentiles were correlated with PLGF and sFlt-1- levels. Results Median birth weight of the newborns from RA patients was 2730 g (range 1250–4000). Among all pregnancies of RA patients, the levels of PLGF and sFlt-1 were similar in active and inactive disease. At the third trimester of pregnancy, RA patients showed lower levels of the pro-angiogenic placental protein PLGF than healthy controls ( p = 0.011). In contrast, levels of the anti-angiogenic protein sFLT-1 did not differ between RA patients and healthy women. The sFlt-1/PLGF ratio was lower in healthy women than in RA patients ( p = 0.020). Moreover the sFlt-1/PLGF ratio correlated negatively with the birth weight percentile. Conclusions RA patients showed reduced levels of the angiogenic placental protein PLGF irrespective of disease activity. The reduced PLGF levels in RA patients might reflect a dysfunction of the placenta which in turn could have an effect on birth weight.

Punica granatum fruit extract inhibits the production of pro-inflammatory cytokines and angiogenic factors of HUVEC cells induced by plasma from patients with pre-eclampsia

Summary This study aims to find out whether the ethanol extract of Punica granatum fruit can inhibit pro-inflammatory cytokines (TNF-α and IL-6) and anti-angiogenic factors (sFlt-1 and sEng) of endothelial cells given plasma stimulus of pre-eclamptic patients. When the cells reached confluence, cells would be grouped into five groups (five replicates per group), i.e. endothelial cells exposed to the plasma of normal pregnancy, endothelial cells exposed to the plasma of patients with severe pre-eclampsia, endothelial cells exposed to the plasma of patients of severe pre-eclampsia and given the extract of P. granatum in various doses (14 ppm; 28 ppm, and 56 ppm). Once induced, measurement of pro-inflammatory cytokine levels and anti-angiogenic factors would be carried out by using the technique of enzyme-linked immunosorbent assay (ELISA). Phytochemical analysis showed that the extract contains phenolic, flavonoid and tannin. Plasma from patients with pre-eclampsia increased the levels of TNF-α, IL-6, sFlt-1 and sEng significantly compared with normal pregnancy plasma exposure group. The increase in the levels of TNF-α, sFlt-1 and sEng was significantly inhibited by the administration of extract at a dose of 56 ppm (P

KL 15 Treatment of very preterm preeclampsia via heparin-mediated extracorporeal LDL-precipitation (H.E.L.P) apheresis results in favorable fetal outcome and weight gain without lowering soluble FMS-like tyrosine kinase-1 (sFlt-1): The Freiburg preeclampsia HELP-apheresis study

Preeclampsia (PE) is a life-threatening pregnancy complication. To date the only therapy for severe PE consists in timely delivery of the fetus. In PE, a deranged lipoprotein metabolism is observed that is believed to impair endothelial function. Heparin-mediated extracorporeal LDL-precipitation (H.E.L.P.)-apheresis is an established method for lipoprotein removal already successfully applied to alleviate PE. Angiogenetic and anti-angiogenetic factors like placenta growth factor (PlGF) and soluble Fms-like tyrosine kinase-1 (sFlt-1) have recently been named as risk factors, and removing sFlt-1 via dextran sulfate column (DSC) apheresis was suggested as a specific cure for PE. By contrast, H.E.L.P.-apheresis does not remove sFlt-1. For the present clinical trial, we revaluated the clinical efficacy of H.E.L.P.-apheresis in PE focusing on sFlt-1 and PlGF. Open pilot study assessing the prolongation by H.E.L.P.-apheresis in 6 women (30–41 years) with very preterm PE (24 + 4 to 27 + 0 gestational weeks (GW)) (NCT01967355) compared to a historic untreated PE-group matching treated patients by GW at admission (<28 GW; n = 6). Clinical outcome of mothers and babies, pre- and post H.E.L.P.-apheresis levels of sFlt-1, PlGF, as well as lipoprotein levels were monitored. Women with PE underwent apheresis 2–6 times. In the treated patients, average time from admission to birth was 15.0 days compared to 6.3 days in controls (p = 0.027). In treated patients the prolongation of gestation by apheresis converted to a mean fetal weight gain of 101 g (p = 0.029). Lung maturation was induced in all treated cases, and all children were released in healthy condition. Apheresis significantly reduced levels of total cholesterol (−33%), triglycerides (−43%), and LDL-cholesterol (−44%). However, sFlt-1 significantly increased initially via H.E.L.P.-apheresis, dropping to almost pre-apheresis levels before the next apheresis, with the sFlt-1/PLGF ratio remaining unaffected. H.E.L.P.-apheresis proved again to be a safe therapeutic option to prolong pregnancies in PE and results in significant fetal weight gain. However, H.E.L.P.-apheresis does not reduce circulating sFlt-1 levels. Lowering lipids and other pro-inflammatory and rheologic factors may thus be more relevant than reducing circulating sFlt-1.

P 59 The sFlt-1/PlGF-ratio an remaining pregnancy duration in RN patients with suspected preeclampsia &gt;34 weeks of gestation

Abstract Introduction The ratio of the antiangiogenic soluble fms-like tyrosine kinase-1 (sFlt-1) and the angiogenic placental growth factor (PlGF) has proven to be a accurate marker for the prediction and diagnosis of pre-eclampsia. The established cut-offs of the ratio provide precise prognoses for the occurrence of especially early-onset pre-eclampsia. However its predictive and diagnostic value is significantly lower in the late-onset disease. Objective To evaluate, whether a high sFlt-1/PlGF-ratio correlates with higher rates of adverse maternal and perinatal outcomes and a shorter remaining time to delivery in patients with late-onset pre-eclampsia. Patients and methods We retrospectively assessed 176 women with suspected pre-eclampsia, bearing a singleton pregnancy, whose first measurement of the sFlt-1/PlGF-ratio took place after 336/7 weeks of gestation. The plasma levels of the ratio were examined for an association with time to delivery as well as adverse maternal and perinatal outcomes until discharge from the hospital. Results We were able to show correlations between the level of sFlt-1/PlGF, adverse maternal and perinatal outcomes and time of delivery. The remaining time to delivery became significantly shorter with a rising sFlt-1/PlGF-ratio ( r = 0,477; p p Conclusion The sFlt-1/PlGF ratio was inversely correlated with the remaining time to delivery and positively correlated with the incidence of maternal and perinatal adverse outcomes. Validating existing cut-offs predicting the remaining time to delivery and finding a specific cut-off on the basis of these data will be the aim of further analyses.

OP 43 The predictive power of placental biomarkers regarding the postpartal course after pre-eclamptic pregnancies

Objective To assess whether placental biomarker maternal serum levels (sFlt-1,PlGF) are predictive for the postpartal course in cases of pre-eclampsia. Material and method We examined a patient cohort of 30 pre-eclamptic cases attending the labor ward in our university hospital. Placental biomarker levels were assessed together with routine lab and clinical parameters prior to delivery, 2 h, 1 day, 2 days, 3 days, 4 days and 6 weeks after delivery. We examined sFlt-1 and PlGF using Human sFlt-1 und PlGF ELISA Kit (R&D). The correlation with different postpartal courses were performed using Chi-Square test, Fischer Exact Test and Mann–Whitney-U-Test. A p-value 0.05 was considered to be statistically significant. The statistical analysis was performed using SPSS 20,0 (SPSS Inc., Chicago, IL). Results 73% of the patients had no clinical and/or laboratory improvement during the postpartal course. The cumulative postpartal course could not be predicted using the palcental biomarkers. Several clinical and laboratory parameters could be predicted using the biomarkers. Elevated liver enzymes were predicted by high sFlt-1 levels 2 h after dleivery and sFlt-1/PlGF Ratio on day 1 after delivery. A persistent high blood pressure correlated significantly with high sFlt-1 levels 2 h after delivery and elevated PlGF and sFlt-1/Ratio on day 1 after delivery. Reduced renal functions correlated with high sFlt-1 and PlGF levels throughout the whole postpartal period. The predictive power of Sflt-1 and PlGF regarding unfavourable outcome within 24 h was evaluated using the CHI-Square-Test. A rising PlGF level on day predicted increased liver enzymes. A rising sFlt-1 on day 2 was able to predict an elevated ASt and average blood pressure. Conclusion A cumulative worsening of postpartal clinical and laboratory course in pre-eclamptic patients was not predicted using the placental biomarkers. Yet, some clinical and laboratory parameters correlated with these biomakers. Given a possible different pathomechanism of each grade of pre-eclampsia, we think a higher of patients is needed for further evaluation.

KL 37 Statins for prevention of pre-eclampsia

Pre-eclampsia is associated with serious maternal and perinatal morbidity and in low resource nations accounts for over 20% of all maternal and perinatal mortality. An effective prophylaxis or targeted therapy against pre-eclampsia is desperately needed. Statins inhibit sFlt-1 secretion from endothelial and trophoblast cells in vitro and increase PlGF levels in vivo. Statins may therefore offer a targeted and non-invasive prophylaxis against pre-eclampsia. Previously however, statins taken during pregnancy, especially lipophilic statins that cross plasma membranes, have been linked with major congenital malformations. After adjustment for underlying maternal diabetes however, a large subsequent registry showed no such association between first trimester statin use and congenital anomalies. Nevertheless, uncertainty remains about the safety of statins used after the first trimester on long-term childhood development. We investigated whether statins might offer a potential therapy for pre-eclampsia by conducting the StAmP trial, Statins to Ameliorate Pre-Eclampsia. This double blind multicenter, placebo-controlled randomized trial assessed the effect of a hydrophilic statin Pravastatin on s-FLT1 levels in women with early onset pre-eclampsia. The preliminary results of Stamp and other reports of statins to ameliorate pre-eclampsia will be discussed.