Placental Vascular Endothelial Growth Factor Research Articles

Maternal prediabetes as a risk factor of preeclampsia and placental dysfunction in pregnant female Sprague-Dawley rats

Background Prediabetes (PD) is associated with intermediate hyperglycaemia, dyslipidaemia, reduced nitric oxide (NO) bioavailability and moderate hypertension. All these factors are risk factor for preeclampsia (PE). However, the effects of the PD on placental function have not been shown. Accordingly, this study sought to investigate a possible link between maternal PD and the risk of developing PE. Methods Pregnant female Sprague-Dawley rats (N = 18) were divided into normal, preeclamptic and prediabetic groups (n = 6 in each group) to study the effects of maternal PD on placenta function over the period of 19 days. Blood glucose and blood pressure were measured on gestational day (GND) 0, 9 and 18. Placental vascular endothelial growth factor (VEGF), placenta growth factor (PlGF) and soluble fms-like tyrosine kinase 1 (sFlt-1) mRNA expression were measured terminally. Data were analysed using ANOVA followed by the Tukey–Kramer post hoc test. Values of p < .05 were used to indicate statistical significance. Results Maternal PD and PE significantly increased blood glucose, decrease NO concentration and increase in MAP by comparison to the normal pregnant control group. Maternal PD significantly decreased VEGF, PlGF mRNA expression with a slight increase in sFlt-1 mRNA expression comparison to the normal pregnant control group. Conclusions Maternal PD is associated with placental dysfunction due to impaired glucose handling, endothelial dysfunction and an imbalance in angiogenic and antiangiogenic factors. Therefore, maternal PD is a risk factor of PE.

MiR-424-5p Inhibits Proliferation, Migration, Invasion and Angiogenesis of the HTR-8/SVneo Cells Through Targeting LRP6 Mediated β-catenin.

The aim of this study was to investigate the effects of miR-424-5p on biological behaviors and angiogenesis of the HTR-8/SVneo Cells. Our study included 60 parturient women, which were divided into an PA group (placenta accreta, n = 30) and a normal group (normal placenta, n = 30). QPCR was used to measure the expression of miR-424-5p in placental tissues. The effects of the miR-424-5p mimic on proliferation, migration, and invasion of human HTR-8/SVneo cells and angiogenesis were analyzed. The potential modulated relationship between miR-424-5p and low-density lipoprotein receptor-related protein-6 (LRP6) was demonstrated by luciferase assay. The expression of LRP6, β-catenin, matrix metalloproteinase-2 (MMP-2), placental growth factor (PGF) and vascular endothelial growth factor (VEGF) were measured by qPCR and Western blot assays. The expression of miR-424-5p in the PA group was significantly decreased than that in the normal group. The expression of miR-424-5p has negative correlation with blood loss. Upregulation of miR-424-5p significantly suppressed the cell proliferation, migration, and invasion of HTR-8/SVneo cells in vitro, as well as the tube formation of human umbilical vein endothelial cells (HUVECs). The luciferase assay demonstrated that LRP6 was a target of miR-424-5p. The expression of LRP6, β-catenin, MMP-2, PGF and VEGF were also decreased with upregulation of miR-424-5p (p < 0.05). The inhibitory effects of miR-424-5p on HTR-8/SVneo cells and angiogenesis were enhanced by downregulation of LRP6, but were reversed by upregulation of LRP6. The present study suggests that downregulation of miR-424-5p is related to the occurrence of PA. Enhancing miR-424-5p inhibits proliferation, migration, invasion and angiogenesis of the HTR-8/SVneo cells through targeting LRP6 mediated β-catenin, providing more insights about PA.

High-fat, high-carbohydrate diet-induced prediabetes preconception in Sprague-Dawley rats as a risk factor for the development of preeclampsia: assessing changes in placental metabolic insults.

Hyperglycemia preconception deranges the establishment of a functional placenta; however, the risk of developing preeclampsia (PE) in prediabetic patients remains obscure. The aim was to assess abnormal placental changes as a risk factor for the development of PE in high-fat, high-carbohydrate (HFHC) diet-induced prediabetic (PD) rats. HFHC diet-induced female prediabetic Sprague-Dawley rats were mated, and blood glucose concentrations, mean arterial pressure (MAP), and body weights were monitored on gestational days (GNDs) 0, 9, and 18. On GND 18, animals were euthanized. Blood and placentas were collected for biochemical analysis. Prediabetic rats showed significantly increased blood glucose concentration, proinflammatory cytokines, MAP, placental weight, and fetoplacental ratio compared with non-prediabetic (NPD) rats. Prediabetic rats showed significantly decreased placental vascular endothelial growth factor receptor 1 (VEGFR1) and placental growth factor (PLGF) and plasma nitric oxide (NO) compared with NPD. Prediabetes may have promoted endothelial dysfunction in the placenta and hypoxia, thus reducing PLGF and VEGFR1, which may have promoted proinflammation, endothelial dysfunction associated with NO decline, and hypertension, which is also observed in preeclamptic patients. Prediabetes may have promoted lipogenesis in placentas and fetuses that may have induced macrosomia and IUGR, also observed in preeclamptic patients. The findings from this study highlight the need for screening and monitoring of prediabetes during pregnancy to reduce the risk of developing preeclampsia.

Comparison of angiogenic potential in vitrified vs. slow frozen human ovarian tissue

Vitrification of ovarian tissue is a promising alternative approach to the traditional slow freezing method. Few empirical investigations have been conducted to determine the angiogenic profiles of these two freezing methods. In this study we aimed to answer the question whether one of the cryopreservation methods should be preferred based on the secretion of angiogenic factors. Tissue culture with reduced oxygen (5%) was conducted for 48 h with samples of fresh, slow frozen/thawed and vitrified/rapid warmed ovarian cortex tissue from 20 patients. From each patient, tissue was used in all three treatment groups. Tissue culture supernatants were determined regarding cytokine expression profiles of angiogenin, angiopoietin-2, epidermal growth factor, basic fibroblast growth factor, heparin binding epidermal growth factor, hepatocyte growth factor, Leptin, Platelet-derived growth factor B, placental growth factor and vascular endothelial growth factor A via fluoroimmunoassay. Apoptotic changes were assessed by TUNEL staining of cryosections and supplemented by hematoxylin and eosin and proliferating cell nuclear antigen staining. Comparing the angiogenic expression profiles of vitrified/rapid warmed tissue with slow frozen/thawed tissue samples, no significant differences were observed. Detection of apoptotic DNA fragmentation via TUNEL indicated minor apoptotic profiles that were not significantly different comparing both cryopreservation methods. Vitrification of ovarian cortical tissue does not appear to impact negatively on the expression profile of angiogenic factors and may be regarded as an effective alternative approach to the traditional slow freezing method.

Deficiency of hydrogen sulfide production and pregnancy rate in an experimental model: Association with preterm delivery.

Pro-inflammatory phenomena drive preterm delivery (PTD). Hydrogen sulfide is a gasotransmitter with anti-inflammatory properties produced through the activity of the enzyme cystathionine-γ-lyase (CSE), and its impact was studied in models of normal delivery and PTD in mice. Female CSE+/+ and CSE-/- mice were mated with male CSE+/+ mice; mating was done with drinking water unsupplemented and supplemented with cysteine. The pregnancy rate was monitored. PTD was induced by the intraperitoneal injection of bacterial lipopolysaccharide (LPS) on day 14.5 of pregnancy. Mice were sacrificed for tissue collection and splenocyte isolation after 6 and 12h. Isolated splenocytes were stimulated for the production of tumor necrosis factor-alpha (TNFα), interleukin (IL)-10 and interferon-gamma (IFNγ); TNFα and vascular endothelial growth factor (VEGF) were measured in the fetuses and the placenta. The successful pregnancy rate was lower in CSE-/- mice and it was restored with cysteine supplementation. CSE deficiency was associated with higher tissue concentrations of TNFα in the fetuses, attenuated IL-10 responses and higher IFNγ production from splenocytes. CSE deficiency was not associated with PTD. Following PTD induction, CSE-/- mice did not show attenuated IL-10 responses but the production of TNFα and IFNγ was lowered over-time; placental VEGF was also increased over-time. CSE deficiency has an unfavorable impact on pregnancy. H2 S deficiency through CSE does not drive PTD but mediates pro-inflammatory phenomena in fetuses.

Unveiling Distribution of Per- and Polyfluoroalkyl Substances in Matched Placenta-Serum Tetrads: Novel Implications for Birth Outcome Mediated by Placental Vascular Disruption.

The placenta is pivotal for fetal development and maternal-fetal transfer of many substances, including per- and polyfluoroalkyl substances (PFASs). However, the intraplacental distribution of PFASs and their effects on placental vascular function remain unclear. In this study, 302 tetrads of matched subchorionic placenta (fetal-side), parabasal placenta (maternal-side), cord serum, and maternal serum samples were collected from Guangzhou, China. Eighteen emerging and legacy PFASs and five placental vascular biomarkers were measured. Results showed that higher levels of perfluorooctanoic (PFOA), perfluorooctane sulfonic acid (PFOS), and chlorinated polyfluorinated ether sulfonic acids (Cl-PFESAs) were detected in subchorionic placenta compared to parabasal placenta. There were significant associations of PFASs in the subchorionic placenta, but not in the serum, with placental vascular biomarkers (up to 32.5%) and lower birth size. Birth weight was negatively associated with PFOA (β: -103.8, 95% CI: -186.3 and -21.32) and 6:2 Cl-PFESA (β: -80.04, 95% CI: -139.5 and -20.61), primarily in subchorionic placenta. Mediation effects of altered placental angiopoietin-2 and vascular endothelial growth factor receptor-2 were evidenced on associations of adverse birth outcomes with intraplacental PFOS and 8:2 Cl-PFESA, explaining 9.5%-32.5% of the total effect. To the best of our knowledge, this study is the first to report on differential intraplacental distribution of PFASs and placental vascular effects mediating adverse birth outcomes and provides novel insights into the placental plate-specific measurement in PFAS-associated health risk assessment.

Increased Expression Levels of Soluble Fms-Like Tyrosine Kinase-1 Inhibit the Development of the Nervous System

Introduction: Preeclampsia (PE) remains a leading cause of maternal and perinatal morbidity. At present, only limited options are available for the treatment of PE. Consequently, many patients need to terminate their pregnancies to relieve the disease. Soluble fms-like tyrosine kinase-1 (sFlt-1) is a decoy receptor of placental growth factor and vascular endothelial growth factor which can promote angiogenesis. Throughout pregnancy, the expression level of sFlt-1 continues to increase in both the mother with PE and her offspring. Material and Methods: In this experiment, we generated a zebrafish line expressing high levels of sFlt-1 and investigated changes in behavior and development of the nervous system. Results: At 96 h post-fertilization (hpf), the brain volume area of zebrafish in the experimental group (zFLT1+CasRx) was significantly smaller after injection than in the WT group (p < 0.05) and the negative control group (CasRx) (p < 0.05). At 96 hpf, compared with the WT group, the cerebral blood vessels in the CasRx control group and experimental group (zFLT1-sgRNA+CasRx) were significantly lower after injection (p < 0.05). Compared with the CasRx control group, the track movement distance and the mean track speed of zebrafish in the experimental group (zFLT1-sgRNA+CasRx) after the 6th injection were significantly decreased (p < 0.05). Conclusions: The increased expression levels of sFlt-1 in zebrafish inhibited the development of the cerebral blood vessels, influenced brain volumes, and inhibited behavioral activities. Our data suggest that the elevation of sFlt-1 in the pathological state of PE can inhibit the development of the nervous system in offspring.

Vitreous protein networks around ANG2 and VEGF in proliferative diabetic retinopathy and the differential effects of aflibercept versus bevacizumab pre-treatment

Extracellular signalling proteins interact in networks rather than in isolation. In this context we investigated vitreous protein levels, including placental growth factor (PlGF), angiopoietin-2 (ANG2) and vascular endothelial growth factor (VEGF), in patients with proliferative diabetic retinopathy (PDR) with variable disease severities, and after anti-VEGF pre-treatment. Vitreous samples of 112 consecutive patients undergoing vitrectomy for PDR and of 52 non-diabetic patients with macular holes as controls were studied. A subset of the PDR patients were treated with either aflibercept (AFB, n = 25) or bevacizumab (BVZ)/ranibizumab (RZB) (n = 13), before surgery. Antibody-based analysis of 35 proteins (growth factors and cytokines) showed a significant increase in expression levels of 27 proteins in PDR patients as compared to controls. In network analysis of co-regulated proteins, a strong correlation in expression levels between VEGF, PlGF, MCP1 and ANG2 was found, mostly clustered around ANG2. In the AFB treatment group, concentrations of several proteins were decreased, including VEGFR1, whereas interleukin 6 and 8 were increased as compared to untreated PDR patients. The observed differences in vitreous protein levels between the different treatments and untreated PDR patients may underlie differences in clinical outcomes in patients with PDR.

Hydroxychloroquine reduces soluble Flt-1 secretion from human cytotrophoblast, but does not mitigate markers of endothelial dysfunction in vitro.

Preeclampsia is a multi-system disease that can have severe, even fatal implications for the mother and fetus. Abnormal placentation can lead to ischaemic tissue injury and placental inflammation. In turn, the placenta releases anti-angiogenic factors into the maternal circulation. These systemically act to neutralise angiogenic factors causing endothelial dysfunction causing preeclampsia. Hydroxychloroquine is an immune modulating drug that is considered safe in pregnancy. There is epidemiological evidence suggesting it may reduce the risk of preeclampsia. Here, we examined the effects hydroxychloroquine on the production and secretion of sFlt-1, soluble endoglin (sENG), placental growth factor (PlGF) and vascular endothelial growth factor (VEGF) in primary human placenta, cytotrophoblasts and umbilical vein endothelial cells (endothelial cell model). Hydroxychloroquine treatment decreased mRNA expression of two sFlt-1 isoforms and its protein secretion. sENG was not reduced. Hydroxychloroquine treatment increased secretion of pro-angiogenic factor PIGF from endothelial cells. It did not significantly reduce the expression of the endothelial cell inflammation marker, ET-1, and inflammation induced expression of the adhesion molecule, VCAM. Hydroxychloroquine could not overcome leukocyte adhesion to endothelial cells. Hydroxychloroquine mitigates features of preeclampsia, but it does not reduce key markers of endothelial dysfunction.

Role of progesterone on dexamethasone-induced alterations in placental vascularization and progesterone receptors in rats†.

Intrauterine growth restriction (IUGR) is manifested by lower maternal progesterone levels, smaller placental size, and decreased placental vascularity indicated by lower expression of vascular endothelial growth factor (VEGF). Studies showed that progesterone increases angiogenesis and induces VEGF expression in different tissues. Therefore, the aim of the present study is to evaluate the effect of progesterone on placental vascular bed and VEGF expression and the modulation of nuclear and membranous progesterone receptors (PR) in dexamethasone-induced rat IUGR model. Pregnant Sprague-Dawley rats were allocated into four groups and given intraperitoneal injections of either saline, dexamethasone, dexamethasone, and progesterone or progesterone. Injections started on gestation day (DG) 15 and lasted until the days of euthanization (19 and 21 DG). Enzyme-linked immunosorbent assay was used to evaluate plasma progesterone levels. Real-time PCR and western blotting were used to evaluate gene and protein expressions of VEGF, and PR in labyrinth and basal placental zones. Immunohistochemistry was used to locate VEGF and different PRs in placental cells. Immunofluorescence was used to monitor the expression of blood vessel marker (αSMA). Dexamethasone decreased the vascular bed fraction and the expression of VEGF in both placental zones. Progesterone co-treatment with dexamethasone prevented this reduction. Nuclear and membrane PRs showed tissue-specific expression in different placental zones and responded differently to both dexamethasone and progesterone. Progesterone treatment improves the outcomes in IUGR pregnancy. Progesterone alleviated DEX-induced IUGR probably by promoting placental VEGF and angiogenesis.

Evaluation of the Effect of the Fibroblast Growth Factor Type 2 (FGF-2) Administration on Placental Gene Expression in a Murine Model of Preeclampsia Induced by L-NAME.

The abnormal implantation of the trophoblast during the first trimester of pregnancy precedes the appearance of the clinical manifestations of preeclampsia (PE), which is a hypertensive disorder of pregnancy. In a previous study, which was carried out in a murine model of PE that was induced by NG-nitro-L-arginine methyl ester (L-NAME), we observed that the intravenous administration of fibroblast growth factor 2 (FGF2) had a hypotensive effect, improved the placental weight gain and attenuated the fetal growth restriction, and the morphological findings that were induced by L-NAME in the evaluated tissues were less severe. In this study, we aimed to determine the effect of FGF2 administration on the placental gene expression of the vascular endothelial growth factor (VEGFA), VEGF receptor 2 (VEGFR2), placental growth factor, endoglin (ENG), superoxide dismutase 1 (SOD1), catalase (CAT), thioredoxin (TXN), tumor protein P53 (P53), BCL2 apoptosis regulator, Fas cell surface death receptor (FAS), and caspase 3, in a Sprague Dawley rat PE model, which was induced by L-NAME. The gene expression was determined by a real-time polymerase chain reaction using SYBR green. Taking the vehicle or the L-NAME group as a reference, there was an under expression of placental VEGFA, VEGFR2, ENG, P53, FAS, SOD1, CAT, and TXN genes in the group of L-NAME + FGF2 (p < 0.05). The administration of FGF2 in the murine PE-like model that was induced by L-NAME reduced the effects that were generated by proteinuria and the increased BP, as well as the response of the expression of genes that participate in angiogenesis, apoptosis, and OS. These results have generated valuable information regarding the identification of molecular targets for PE and provide new insights for understanding PE pathogenesis.

Pathophysiology of Placenta in Antiphospholipid Syndrome

Antiphospholipid syndrome (APS) is an autoimmune disorder characterized by clinical manifestations caused by arterial or venous thrombosis and pregnancy conditions such as recurrent miscarriage, fetal death, or premature birth in the presence of antiphospholipid antibodies. The obstetrical manifestations are strongly related to the placental alterations. The aim of this review is to summarize the latest data on pathophysiology of obstetrical APS, emphasizing the disturbance of the placentation process. Due to a lack of extravillous trophoblasts to properly reconstruct the spiral arteries, APS causes hypoxic or ischemic injury or high-speed blood flow that damages the placenta. This results in decreased or interrupted maternal blood flow to the placenta and a lack of nutrients for the fetus. Antiphospholipid antibodies can lower the proliferation and infiltration of the extravillous trophoblasts. The placental mal-perfusion causes the release of antiangiogenic substances such as soluble fms-like tyrosine kinase-1 and soluble endoglin. Placental growth factor and vascular endothelial growth factor (VEGF) may be sequestered by sFlt1 and blocked from binding to trophoblast and endothelial cell VEGF receptors, inhibiting their proangiogenic effects. Preeclampsia is the clinical result from a lack of angiogenic factors needed for endothelial vascular homeostasis due to an excess of sFlt1 in the maternal circulation.

Omega-3 and Vitamin E Supplementation Effect on Reactive Oxygen Species and Placental Vascular Endothelial Growth Factor: Pre-eclampsia Model

BACKGROUND: Pre-eclampsia (PE) has been associated with oxidative stress and inflammation. Omega-3 fatty acids and vitamin E have beneficial function to maintain cell membrane, prevent oxidative stress, and inhibit the production of proinflammatory cytokines. The purpose of the study was to determine the effect of omega-3 and vitamin E supplement to Reactive Oxygen Species (ROS) and placental Vascular Endothelial Growth Factor (VEGF) on PE rats model. AIM: This study aimed to investigate the effect of omega-3 and vitamin E supplementation on ROS and placental VEGF. METHODS: This research has been carried out at animal house and Biomedical Laboratory of the Medical Faculty Andalas University. The design of this research was experimental study with post-test only control group design. Thirty pregnant rats were divided into five groups. Group K-without treatment, group K+ was given L-NAME, group P1 was given L-NAME + omega-3, group P2 was given L-NAME + vitamin E, and group P3 was given L-NAME + omega-3 + vitamin E. The L-NAME dose was 50 mg/kg/day. At the 19th day of gestation, the pregnancy was terminated and the blood serum was used for examination of ROS and placental tissue was collected for examination of VEGF using ELISA kit. The data were analyzed by one-way ANOVA and post hoc test Bonferoni. RESULTS: The analysis results found that there were significant differences between omega-3 and vitamin E supplements on ROS levels and placental VEGF levels. Administration of omega-3 only or combination with vitamin E can effectively improve endothelial function by increasing the level of VEGF. CONCLUSION: Omega-3 (DHA 120 mg, EPA 180 mg) and vitamin E (α-tocopherol 300 iu) supplement decreased ROS and increased placental VEGF on pre-eclampsia rats model.

Gq Signaling in the Placental Syncytiotrophoblast Layer During Preeclampsia

Preeclampsia is a pregnancy‐associated cardiovascular disorder characterized by hypertension and proteinuria after 20 weeks of gestation. Generalized cellular ‘stress’ (oxidative, mitochondrial, etc.) within the syncytiotrophoblast (STB) layer of placenta is theorized to drive its pathogenesis. Hormones implicated in preeclampsia such as angiotensin, endothelin, and vasopressin signal via receptors that typically couple to the Gq second‐messenger cascade, and Regulator of G protein Signaling‐2 (RGS2) buffers this signaling. We have published that RGS2 expression is decreased in human preeclamptic placenta, and reducing RGS2 expression in placenta is sufficient to cause development of key features of this disorder in mice. Unpublished in situ hybridization data indicate that in both humans and mice, RGS2 is present in the STB layer – and that RGS2 expression in the STB layer is reduced during preeclampsia in humans. Collectively, these findings lead us to generally hypothesize that excess Gq signaling specifically within the STB layer, whether due to excessive hormonal activation or loss of RGS2‐mediated buffering, causes STB‐Stress and thereby contributes to the pathogenesis of preeclampsia. Therefore, the objective of this study was to test whether artificial, exogenous activation of Gq signaling only within STB cells is sufficient to induce placental and maternal phenotypes of preeclampsia in mice.CAG‐FLEX‐hM3Dq+/+ (Gαq DREADD, Jax 026943) or C57BL/6J dams were bred with Gcm1‐Cre+/‐ sires (Jax 026625) or non‐transgenic littermates to elicit STB‐restricted hM3Dqexpression in ≍50% or 0% of placentas. The synthetic DREADD ligand clozapine N‐oxide (CNO) or saline vehicle was administered (2 mg/kg/d, ip) mid‐gestation (GD 12.5‐14.5). Placental and maternal phenotypes were compared between Cre– and Cre+ placentas within a dam, CNO‐ vs saline‐injected double‐transgenic dams, and double‐ vs single/non‐transgenic dams treated with CNO. Activation of Gq in the STB layer caused maternal proteinuria (n=4‐9, 49±6 mg/day vs 28±4 mg/day; p=0.01). Labyrinth vascularization, assessed by percentage of CD31+ area, was reduced with Gq activation (n=5, Cre+ 21±1%, Cre– 27±2%; p=0.01), while other regions remained unaffected (Decidua: Cre+ 14±4%, Cre– 14±3%; Junctional Zone: Cre+ 11±4%, Cre– 12±3%). Possible reductions in placental mass (n=5 Cre+ 0.12±0.01g, Cre– 0.15±0.01g; p=0.06), and placental vascular endothelial growth factor protein levels (n=6‐7 117±67 ng/g vs 151±12 ng/g; p=0.06) were noted, but no obvious trend in fetal growth restriction was observed (n=5 Cre+ 0.21±0.06g, Cre– 0.29±0.04g). These data support the pathological significance of excess Gq signaling specifically within the STB layer. Ongoing studies are aimed at further characterizing maternal phenotypes (blood pressure, circulating factors) in these models, and the synergistic effects of simultaneous STB‐specific deletion of RGS2 upon sensitivity to Gq stimulation in this cell type.

Shikonin alleviates choroidal neovascularization by inhibiting proangiogenic factor production from infiltrating macrophages

Choroidal neovascularization (CNV), a feature of neovasular age-related macular degeneration (AMD), acts as a leading cause of vision loss in the elderly. Shikonin (SHI), a natural bioactive compound extracted from Chinese herb radix arnebiae, exerts anti-inflammatory and anti-angiogenic roles and also acts as a potential pyruvate kinase M2 (PKM2) inhibitor in macrophages. The major immune cells macrophages infiltrate the CNV lesions, where the production of pro-angiognic cytokines from macrophage facilitates the development of CNV. PKM2 contributes to the neovascular diseases. In this study, we found that SHI oral gavage alleviated the leakage, area and volume of mouse laser-induced CNV lesion and inhibited macrophage infiltration without ocular cytotoxicity. Moreover, SHI inhibited the secretion of pro-angiogenic cytokine, including basic fibroblast growth factor (FGF2), insulin-like growth factor-1 (IGF1), chemokine (C-C motif) ligand 2 (CCL2), placental growth factor and vascular endothelial growth factor (VEGF), from primary human macrophages by down-regulating PKM2/STAT3/CD163 pathway, indicating a novel potential therapy strategy for CNV.

Macular Microvascular Changes and Their Correlation With Peripheral Nonperfusion in Branch Retinal Vein Occlusion

To investigate the correlation between macular microvascular alterations on optical coherence tomography angiography (OCTA) and retinal ischemia on ultra-widefield fluorescein angiography (UWF FA) in eyes with branch retinal vein occlusion (BRVO). Cross-sectional study. This prospective study was performed from September 2019 to June 2020 at Yeungnam University Medical Center. We included 60 patients with treatment-naïve BRVO. Two independent, masked graders analyzed OCTA parameters, including vessel density, skeletal density, and fractal dimension (FD), and UWF FA parameters, including retinal nonperfusion area (NPA) and ischemic index (ISI), from various concentric regions (perimacular region, 0.5-3mm radius; near-peripheral region, 3-10mm; midperipheral region, 10-15mm; far-peripheral region, >15mm). A repeated-measures analysis of variance test and a paired t test were performed for inter-visit and inter-regional comparisons, and Pearson correlation coefficient and multivariate regression analyses were performed to examine the correlation between UWF FA and OCTA parameters. The OCTA parameters from both the superficial and deep capillary plexuses (DCP) were significantly correlated with NPA and ISI in all concentric regions. Even after adjusting for several covariates, all OCTA parameters revealed a significant association with ISI on UWF FA. Moreover, OCTA parameters from DCP were significantly correlated with concentrations of placental growth factor and vascular endothelial growth factor. Although all OCTA parameters achieved excellent results of area under the curve (AUC) > 0.9 for detecting severe retinal ischemia, defined as ISI >10%, FD reduction in DCP was the most reliable parameter (AUC= 0.948, P < .001), and 5.39% was the best cut-off point for predicting ISI > 10%. OCTA is a useful noninvasive tool not only for evaluation of macular microvasculature but for supposition of peripheral nonperfusion in eyes with BRVO.

The effect of combination training program on the expression of placental growth factor and vascular endothelial growth factor receptor 1 indicators among coronary artery bypass graft patients

Aims: The aim of this study was the evaluation of combination training program influence on the placental growth factor (PIGF) and vascular endothelial growth factor receptor 1 (VEGFR-1) indicators among CABG patients.Setting and Design: The study method was semi-experimental; the statistical population was some patients who had heart disease, after CABG at Babol city hospitals, Iran.Materials and Methods: 16 patients were selected by convenience sampling method.The expression levels of the PIGF and VEGFR-1 genes were measured by the Real Time PCR method.Statistical Analysis Used: The covariance analysis method and the dependent t-test were used.Results: The results showed that changes in PIGF and VEGFR-1 gene expression in the experimental group had a significant decrease in the posttest stage compared to the pretest stage; While changes in PIGF and VEGFR-1 gene expression in the control group were not significantly different in the posttest stage compared to the pretest stage; also, data analysis showed that changes in gene expression of PIGF and VEGFR-1 factors had a significant decrease in the experimental group compared to the control group.Conclusion: The study results showed 8 weeks of combination exercises among the experimental group had a significant decrease in the expression levels of PIGF and VEGFR1 genes among CABG patients compared to the control group. Physical activity may decrease PIGF levels, which is an important factor in oxidative stress and inflammation.

Increased Expression Levels of Soluble Fms-Like Tyrosine Kinase-1 Inhibits Nervous System Development of Zebrafish

Introduction Preeclampsia (PE) remains a leading cause of maternal and perinatal morbidity. At present, the treatments of preeclampsia are limited, and many patients need to terminate the pregnancy to relieve the disease. Soluble fms-like tyrosine kinase-1 (sFlt-1) is a decoy receptor of placental growth factor (PIGF) and vascular endothelial growth factor which can promote angiogenesis. Its level increases throughout pregnancy with preeclampsia and umbilical cord blood of their newborns. Material and methods In this experiment, we constructed zebrafish with high expression levels of sFlt-1 and explored their changes of nervous system developments and behaviors. Results Comparing with the WT group and the casRx group, the cerebral vascular developments at 72 h in the F1、F2 group were significantly decreased, the ratios of brain volume area to whole body area at 96 h were significantly decreased, and the behavioral trajectory distances and movement speeds at 6th day were shortened and slowed down. Conclusions The increased expression levels of sFlt-1 in zebrafish can inhibit the developments of cerebral blood vessels, brain volumes and the behavioral activities. It is speculated that elevated sFlt-1 in the pathological state of PE can inhibit the developments of nervous system in offspring.

Combining metformin and sulfasalazine additively reduces the secretion of antiangiogenic factors from the placenta: Implications for the treatment of preeclampsia

IntroductionThe antiangiogenic factors soluble fms-like tyrosine kinase-1 (sFlt-1) and soluble endoglin (sENG) are elevated in preeclampsia and have been implicated in its pathogenesis. We have previously demonstrated metformin and sulfasalazine independently reduce antiangiogenic factor secretion. Here we examined whether combining metformin and sulfasalazine may be more effective than either alone in reducing placental expression and secretion of antiangiogenic and angiogenic factors and the expression of markers of endothelial dysfunction. MethodsWe performed functional experiments using primary human placenta to explore the effect of metformin and sulfasalazine, at lower doses than previously explored, individually and in combination, on sFlt-1 and sENG secretion and placental growth factor (PlGF) and vascular endothelial growth factor (VEGFα) expression. Using primary endothelial cells we induced dysfunction using cytokine tumor necrosis factor-α (TNF-α) and assessed the effect of low dose combination treatment on the expression of vascular cell adhesion molecule-1 (VCAM-1) and Endothelin-1 (a potent vasoconstrictor). ResultsWe demonstrated combination metformin and sulfasalazine was additive in reducing sFlt-1 secretion from cytotrophoblasts and placental explants. Combination treatment was also additive in reducing sENG secretion from placental explants. Furthermore, combination treatment increased cytotrophoblast VEGFα mRNA expression. Whilst combination treatment increased PlGF mRNA expression this was similar to treatment with sulfasalazine alone. Combination therapy reduced TNFα induced endothelin-1 mRNA expression however did not change VCAM expression. DiscussionLow dose combination metformin and sulfasalazine reduced cytotrophoblast sFlt-1 and sENG secretion, increased VEGFα expression and reduced TNFα induced endothelin-1 expression in primary endothelial cells. Combination therapy has potential to treat preeclampsia.

A Step-Wedge in the Biochemical Diagnosis of Preeclampsia

Preeclampsia, a hypertensive disorder of pregnancy, is one of the leading causes of maternal and neonatal morbidity and mortality worldwide. Among the reasons for the great global burden of disease is the poor predictive value of high blood pressure and proteinuria (traditional markers for the disease) for related adverse outcomes (1). Angiogenic biomarkers developed and studied during the past decade are widely recognized as clinically sensitive and specific biomarkers, with excellent predictive values during the third trimester that complement and improve upon clinical signs and symptoms (2). Normally produced in pregnancy, soluble FMS-like tyrosine kinase (sFLT1) is a placentally produced, antiangiogenic protein that mitigates rapidly progressing placental angiogenesis driven by proangiogenic proteins—placental growth factor (PlGF) and vascular endothelial growth factor—in the latter third of gestation. Preeclampsia is characterized by enhanced production of placental sFLT1, which in turn binds to PlGF, causing the free concentration of serum PlGF to fall. Increased circulating sFLT1 and decreased free PlGF antedate the onset of preeclamptic signs and symptoms, allowing for accurate and minimally invasive risk assessment of the systemic vascular damage that characterizes preeclampsia (2). Furthermore, angiogenic biomarkers can be used to differentiate preeclampsia from mimickers of disease, such as chronic hypertension, gestational hypertension, and chronic kidney disease (2). Despite the opportunity for early identification of disease before the onset of signs and symptoms, funding bodies have been reluctant to invest in the head-to-head comparisons of angiogenic biomarkers vs traditional factors, leading to a …