Vascular contraction of umbilical arteries of pregnant women with preeclampsia.

Objective: Potassium channels have an important role in the vascular adaptation during pregnancy and a reduction in the expression of adenosine triphosphate-sensitive potassium channels (Katp) has been linked to preeclampsia. Activation of Katp induces vasodilation; however, no previous study has been conducted to evaluate the effects of the inhibition of these channels in the contractility of preeclamptic arteries. Glibenclamide is an oral antihyperglycemic agent that inhibits Katp and has been widely used in vascular studies. Methods: To investigate the effects of the inhibition of Katp, umbilical arteries of preeclamptic women and women with healthy pregnancies were assessed by vascular contractility experiments, in the presence or absence of glibenclamide. The umbilical arteries were challenged with cumulative concentrations of potassium chloride (KCl) and serotonin. Results: There were no differences between the groups concerning the maternal age and gestational age of the patients. The percentage of smokers, caucasians and primiparae per group was also similar. On the other hand, blood pressure parameters were elevated in the preeclamptic group. In addition, the preeclamptic group presented a significantly higher body mass index. The newborns of both groups presented similar APGAR scores and weights. Conclusion: In the presence of glibenclamide, there was an increase in the KCl-induced contractions only in vessels from the PE group, showing a possible involvement of these channels in the disorder.

The Nitrate-Nitrite-Nitric Oxide Pathway: Potential Role in Mitigating Oxidative Stress in Hypertensive Disorders of Pregnancy.

Hypertensive diseases of pregnancy (HDPs) represent a global clinical challenge, affecting 5-10% of women and leading to complications for both maternal well-being and fetal development. At the heart of these complications is endothelial dysfunction, with oxidative stress emerging as a pivotal causative factor. The reduction in nitric oxide (NO) bioavailability is a vital indicator of this dysfunction, culminating in blood pressure dysregulation. In the therapeutic context, although antihypertensive medications are commonly used, they come with inherent concerns related to maternal-fetal safety, and a percentage of women do not respond to these therapies. Therefore, alternative strategies that directly address the pathophysiology of HDPs are required. This article focuses on the potential of the nitrate-nitrite-NO pathway, abundantly present in dark leafy greens and beetroot, as an alternative approach to treating HDPs. The objective of this review is to discuss the prospective antioxidant role of nitrate. We hope our discussion paves the way for using nitrate to improve endothelial dysfunction and control oxidative stress, offering a potential therapy for managing HDPs.

The interplay between extracellular NAMPT and inflammatory cytokines in preeclampsia.

Preeclampsia (PE) is the major cause of maternal-fetal mortality and morbidity. Its pathophysiology is not elucidated, but there is evidence for the role of visfatin/nicotinamide phosphoribosyl transferase (NAMPT), mainly due to its relation to endothelial dysfunction, a hallmark of PE. However, there is heterogeneous data regarding visfatin/NAMPT in healthy pregnancy (HP) and PE. Therefore, we performed a search on MEDLINE/PubMed using the terms "visfatin and preeclampsia" and "NAMPT and preeclampsia, and we selected 23 original articles: 12 articles reported increased levels in PE compared to HP, only four articles showed lower levels and eight articles did not find differences regarding visfatin/NAMPT in the groups studied. It is widely acknowledged that levels detected in plasma, serum, or placenta can be influenced by the size of the population and sample analyzed, as well as genetic factors. We further discussed the correlations of visfatin/NAMPT with clinical biomarkers in PE and inflammatory pathways. Considering the common inflammatory mechanisms between PE and visfatin/NAMPT, few studies have recently performed serum or plasma dosages. In conclusion, further studies are needed to highlight the potential role of visfatin/NAMPT in the pathophysiology of PE. This will provide comparative evidence to establish it as a biomarker for disease outcomes and treatment.

EGCG, a Green Tea Compound, Increases NO Production and Has Antioxidant Action in a Static and Shear Stress In Vitro Model of Preeclampsia.

Preeclampsia (PE) is a gestational hypertensive disease characterized by endothelial dysfunction. Epigallocatechin-3-gallate (EGCG), the main compound in green tea, is a promising therapeutic target for the disease. By activating eNOS, EGCG increased NO production and exerted an important antioxidant action, but its specific impact in the context of PE remains understudied. The aim of this study is to evaluate the effects of EGCG on endothelial function in static and shear stress in in vitro models of PE. Endothelial cells were incubated with healthy (HP) and preeclamptic (PE) pregnant women's plasma, and the latter group was treated with EGCG. Additionally, NOS (L-NAME) and PI3K protein (LY249002) inhibitors were also used. The levels of NO, ROS, and O2•- were evaluated, as well as the antioxidant potential. These investigations were also carried out in a shear stress model. We found that EGCG increases the NO levels, which were reduced in the PE group. This effect was attenuated with the use of L-NAME and LY249002. Furthermore, EGCG increased the antioxidant capacity of PE, but its action decreased with LY294002. In cells subjected to shear stress, EGCG increased nitrite levels in the PE group and maintained its action on the antioxidant capacity. This is the first study of the effects of EGCG in this experimental model, as well as the investigation of its effects along with shear stress. Our findings suggest that EGCG improves parameters of endothelial dysfunction in vitro, making it a promising target in the search for treatments for the disease.

Missing links in preeclampsia cell model systems of endothelial dysfunction.

Preeclampsia, one of the main hypertensive disorders of pregnancy, is associated with circulating factors released by the ischemic placenta accompanied by systemic endothelial dysfunction. The etiology of preeclampsia remains poorly understood although it is associated with high maternal and fetal mortality and increased cardiovascular disease risk. Most cell model systems used for studying endothelial dysfunction have not taken into account hemodynamic physical factors such as shear-stress forces which may prevent extrapolation of cell data to in vivo settings. We overview the role of hemodynamic forces in modulating endothelial cell function and discuss strategies to reproduce this biological characteristic in vitro to improve our understanding of endothelial dysfunction associated with preeclampsia.

Assessment by ABPM verified the presence of hypertension in patients with self-reported hypertension, pregnant women, as well as differences between ethnicities in women aged 38-39 years in the Ribeirão Preto cohort.

Introduction: Arterial hypertension is a global health problem and one of the main risk factors for cardiovascular diseases (CVD), and therefore for morbidity and mortality among adult men and women. Factors related to obstetric history, family history, sociodemographic characteristics, and lifestyle habits are known determinants of arterial hypertension. Methods: Case-control study of women belonging to the 1978/79 birth cohort conducted in the city of Ribeirão Preto/SP. Sociodemographic data, presence of comorbidities, maternal comorbidities, paternal comorbidities, comorbidities during pregnancy, and biometric and biophysical markers associated with blood pressure measured by 24-h ambulatory blood pressure monitoring (ABPM) were assessed in women aged 38-39 years. We want to study which variables of the previous sentence are related to the presence of hypertension measured by ABPM. Results: Data from 281 women were analyzed. Our results showed that ethnicity, a history of hypertension, and gestational hypertension reported by the women were significantly associated with the presence of hypertension measured by ABPM. Other factors such as marital status, educational level, comorbidities of the woman, paternal or maternal comorbidities, anthropometric measurements or serum levels of cardiovascular markers were not associated with the presence of hypertension measured by ABPM. Conclusion: We conclude that ethnicity, self-reported hypertension, and gestational hypertension are associated with arterial hypertension measured by ABPM.

Prediction and prevention of preeclampsia by physicians in Brazil: An original study.

Background: Considering the worldwide importance of preeclampsia, especially in Brazil, the screening of pregnant women at greater risk of developing the disease and the application of preventive measures are essential. This study aimed to assess the medical performance in this context in Brazil. Methods: A survey was developed to quantify the number of physicians who prescribe acetylsalicylic acid (ASA) and/or calcium for preeclampsia prevention. The survey was sent to all Brazilian obstetricians affiliated to the Brazilian Federation of OBGYN by email and WhatsApp. The survey remained opened for 6 months and included questions about the use of ASA and calcium, as well as about the use of a complementary test to predict preeclampsia. Results: The sample consisted of 360 responding physicians and 100% coverage of responses from physicians from the five different regions of Brazil was obtained. The vast majority of respondents (94.72%) prescribe ASA to prevent preeclampsia, with 80.3% prescribing a dose of 100 mg/day. Calcium is prescribed by 83.9% of the respondents. The majority of the interviewed sample (58.6%) requests uterine artery Doppler imaging to predict preeclampsia and 31.7% do not request any additional test. When the analysis was performed by region, only the northern region differed from the other Brazilian regions regarding the use of ASA and calcium for preeclampsia prevention. While more than 90% of physicians in the other regions prescribe ASA, 40% in the northern region do not use it (p < 0.0001). Regarding calcium, 30% of physicians in northern Brazil do not use the drug for preeclampsia prevention, a percentage that also differs from the other regions where the medication is prescribed by 80 to 90% of physicians (p = 0.021). Conclusions: The vast majority of Brazilian physicians prescribe low-dose aspirin and calcium carbonate to prevent preeclampsia in high-risk pregnant women. In addition to the identification of clinical risk factors, most doctors use Doppler of the uterine arteries as a predictive method. In the northern region of Brazil, physicians use aspirin and calcium less frequently for preventing preeclampsia compared to the rest of the country.

Markers of Endothelial Dysfunction Are Attenuated by Resveratrol in Preeclampsia.

Preeclampsia (PE) is characterized by great endothelial dysfunction, decreased nitric oxide (NO) bioavailability, and higher levels of arginase activity. In the present study, we investigated the potential modulatory effects of trans-resveratrol (RSV) on arginase and endothelial dysfunction biomarkers in endothelial cells exposed to plasma from patients with PE and healthy pregnant (HP) women, and umbilical arteries from patients with PE. Human umbilical vein endothelial cells (HUVECs) were incubated with pooled plasma from 10 HP or 10 PE pregnant women and RSV; umbilical arteries from patients with PE were incubated with RSV; intracellular NO and total reactive oxygen species (ROS) levels were assessed using a probe that interacted with these radicals; total arginase activity was evaluated measuring the urea produced; total antioxidant capacity was measured using the ferric reduction ability power (FRAP) assay; and endothelial dysfunction biomarkers were assessed using qPCR in endothelial cells and umbilical arteries. RSV increased NO levels and decreased total arginase activity in endothelial cells incubated with plasma from patients with PE. In addition, RSV increased total antioxidant capacity and downregulated endothelial dysfunction biomarkers, such as intercellular adhesion molecule-1 (ICAM-1), von Willebrand factor (vWF), and Caspase-3, (CASP-3), in endothelial cells and umbilical arteries from PE patients. RSV treatment positively modulated the L-arginine-NO pathway, decreased arginase activity, and increased antioxidant capacity, in addition to downregulating endothelial dysfunction biomarkers.

Glibenclamide Increases Nitric Oxide Levels and Decreases Oxidative Stress in an In Vitro Model of Preeclampsia.

(1) Background: The bioavailability of nitric oxide (NO) and oxidative stress are important events related to the pathophysiology of preeclampsia (PE). In this present study, we aimed to evaluate the antioxidant effect of glibenclamide (GB) on the NO synthesis, oxidative stress, and antioxidant capacity in endothelial cells incubated with plasma from preeclamptic (PE) and normotensive pregnant women (NT). (2) Methods: Human umbilical vein endothelial cells (HUVECs) were incubated with a plasma pool from 10 NT and 10 PE pregnant women; NO/NOx quantification and ROS levels were assessed by a fluorescence compound; lipid peroxidation was evaluated employing thiobarbituric acid (TBA); and total antioxidant capacity was measured by ferric reduction ability power (FRAP) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). (3) Results: We found that endothelial cells incubated with plasma from PE showed lower NO and NOx levels compared with the NT group. However, GB treatment increased these levels, as well as the antioxidant capacity. Furthermore, a decrease was observed in ROS generation and lipid peroxidation (4) Conclusions: The GB treatment exerted a positive effect on the NO/NOx production by HUVEC incubated with plasma from NT and PE pregnant women, as well as in the reduction in oxidative stress and increase in the antioxidant capacity.

Nebivolol Increases Nitric Oxide Synthase via ß3 Adrenergic Receptor in Endothelial Cells Following Exposure to Plasma from Preeclamptic Patients.

Background: Low bioavailability of nitric oxide (NO) is related to the pathophysiology of preeclampsia (PE). In the present study, we investigated the effect of nebivolol (NEB), a ß3-receptor agonist with vasodilator properties, on the NO synthesis in endothelial cells incubated with plasma from preeclamptic patients. Methods and results: Human umbilical vein endothelial cells (HUVECs) were incubated with plasma from healthy pregnant (HP) and PE women; NO quantification was assessed by a fluorescence compound. We found that endothelial cells incubated with plasma from women with PE show lower NO levels compared with the HP group (p < 0.0001). However, NEB treatment increases NO levels, partially, mediated by ß3 adrenergic receptors (p < 0.0001) and through eNOS activation (p < 0.0001). Conclusions: Our results suggest that NEB acts in NO synthesis through eNOS activation and ß3 adrenergic receptors in the endothelium. However, further studies will be needed to understand this molecule.

Effects of chronic dietary nitrate supplementation on longevity, vascular function and cancer incidence in rats.

RATIONALE: Dietary nitrate and nitrite have a notoriously bad reputation because of their proposed association with disease, in particular cancer. However, more recent lines of research have challenged this dogma suggesting that intake of these anions also possess beneficial effects after in vivo conversion to the vital signaling molecule nitric oxide. Such effects include improvement in cardiovascular, renal and metabolic function, which is partly mediated via reduction of oxidative stress. A recent study even indicates that low dose of dietary nitrite extends life span in fruit flies. METHODS: In this study, 200 middle-aged Wistar rats of both sexes were supplemented with nitrate or placebo in the drinking water throughout their remaining life and we studied longevity, biochemical markers of disease, vascular reactivity along with careful determination of the cause of death. RESULTS: Dietary nitrate did not affect life span or the age-dependent changes in markers of oxidative stress, kidney and liver function, or lipid profile. Ex vivo examination of vascular function, however, showed improvements in endothelial function in rats treated with nitrate. Neoplasms were not more common in the nitrate group. CONCLUSION: We conclude that chronic treatment with dietary nitrate does not affect life span in rats nor does it increase the incidence of cancer. In contrast, vascular function was improved by nitrate, possibly suggesting an increase in health span.

Preeclampsia and Gestational Hypertension: Biochemical and Antioxidant Features in Vitro Might Help Understand Different Outcomes.

OBJECTIVE: Gestational hypertension (GH) is characterized by increased blood pressure after the 20th gestational week; the presence of proteinuria and/or signs of end-organ damage indicate preeclampsia (PE). Heme oxygenase-1 (HO-1) is an antioxidant enzyme with an important role in maintaining endothelial function, and induction of HO-1 by certain molecules shows potential in attenuating the condition's effects over endothelial tissue. HO-1 production can also be stimulated by potassium iodide (KI). Therefore, we evaluated the effects of KI over HO-1 expression in human umbilical vein endothelial cells (HUVECs) incubated with plasma from women diagnosed with GH or PE. METHODS: Human umbilical vein endothelial cells were incubated with a pool of plasma of healthy pregnant women (n = 12), pregnant women diagnosed with GH (n = 10) or preeclamptic women (n = 11) with or without the addition of KI for 24 hours to evaluate its effect on this enzyme expression. Analysis of variance was performed followed by Dunnet's test for multiple comparisons between groups only or between groups with addition of KI (p ≤ 0.05). RESULTS: KI solution (1,000 µM) reduced HO-1 in the gestational hypertension group (p = 0.0018) and cytotoxicity in the preeclamptic group (p = 0.0143); treatment with KI reduced plasma cytotoxicity but did not affect the preeclamptic group's HO-1 expression. CONCLUSION: Our findings suggest that KI alleviates oxidative stress leading to decreased HO-1 expression; plasma from preeclamptic women did not induce the enzyme's expression in HUVECs, and we hypothesize that this is possibly due to inhibitory post-transcriptional mechanisms in response to overexpression of this enzyme during early pregnancy.

OBJETIVO: A hipertensão gestacional (GH) é caracterizada pelo aumento da pressão sanguínea após a 20ª semana de gestação; a presença de proteinuria e/ou sinais de danos a órgãos como rins, fígado e cérebro indicam pré-eclâmpsia (PE). A heme oxigenase-1 (HO-1) é uma enzima antioxidante com um papel importante na manutenção da função endotelial, e a sua indução por certas moléculas se mostra potencialmente benéfica frente à característica deletéria destas condições sobre o endotélio. Já foi demonstrado anteriormente que a produção de HO-1 pode ser induzida por iodeto de potássio (KI). Portanto, nós avaliamos os efeitos do KI sobre a citotoxicidade e expressão de HO-1 por células de veia de cordão umbilical humano (HUVECs) após incubação com o plasma de mulheres diagnosticadas com GH ou PE. MéTODOS: Células de veia de cordão umbilical humano foram incubadas com pool de plasma de gestantes saudáveis (n = 12), gestantes com GH (n = 10) ou gestantes com PE (n = 11) com ou sem a adição de KI por 24 horas para avaliar a citotoxicidade através da dosagem de lactato desidrogenase e produção de HO-1 por ELISA. Foi realizada ANOVA seguida de teste de Dunnet para múltiplas comparações entre os grupos estudados, considerando significativos valores de p ≤ 0,05. RESULTADOS: A solução de KI (1.000 µM) reduziu a produção de HO-1 no grupo GH (p = 0.0018) e a citotoxicidade no grupo PE (p = 0.0143); o tratamento com KI não afetou a produção de HO-1 por HUVECs incubadas com o plasma do grupo PE. CONCLUSãO: Nossos achados sugerem que o KI atenua os efeitos do plasma de gestantes com GH ocasionando a diminuição da produção de HO-1; plasma do grupo PE não induziu a produção de HO-1 em HUVECs em comparação ao grupo saudável, e nossa hipótese é a de que tal achado pode ser devido a mecanismos pós-transcricionais em resposta a uma superestimulação da produção de HO-1 nos estágios iniciais da gravidez.

Preeclampsia and Gestational Hypertension: Biochemical and Antioxidant Features in Vitro Might Help Understand Different Outcomes / Pré-eclâmpsia e hipertensão gestacional: Fatores bioquímicos e antioxidantes in vitro podem auxiliar no entendimento de resultados clínicos distintos

Abstract Objective Gestational hypertension (GH) is characterized by increased blood pressure after the 20th gestational week; the presence of proteinuria and/or signs of end-organ damage indicate preeclampsia (PE). Heme oxygenase-1 (HO-1) is an antioxidant enzyme with an important role in maintaining endothelial function, and induction of HO-1 by certain molecules shows potential in attenuating the condition's effects over endothelial tissue. HO-1 production can also be stimulated by potassium iodide (KI). Therefore, we evaluated the effects of KI over HO-1 expression in human umbilical vein endothelial cells (HUVECs) incubated with plasma from women diagnosed with GH or PE. Methods Human umbilical vein endothelial cells were incubated with a pool of plasma of healthy pregnant women (n = 12), pregnant women diagnosed with GH (n = 10) or preeclamptic women (n = 11)with or without the addition of KI for 24 hours to evaluate its effect on this enzyme expression. Analysis of variance was performed followed by Dunnet's test for multiple comparisons between groups only or between groups with addition of KI (p ≤ 0.05). Results KI solution (1,000 µM) reduced HO-1 in the gestational hypertension group (p = 0.0018) and cytotoxicity in the preeclamptic group (p = 0.0143); treatment with KI reduced plasma cytotoxicity but did not affect the preeclamptic group's HO-1 expression. Conclusion Our findings suggest that KI alleviates oxidative stress leading to decreased HO-1 expression; plasma from preeclamptic women did not induce the enzyme's expression in HUVECs, and we hypothesize that this is possibly due to inhibitory post-transcriptional mechanisms in response to overexpression of this enzyme during early pregnancy.

Resumo Objetivo A hipertensão gestacional (GH) é caracterizada pelo aumento da pressão sanguínea após a 20ª semana de gestação; a presença de proteinuria e/ou sinais de danos a órgãos como rins, fígado e cérebro indicam pré-eclâmpsia (PE). A heme oxigenase-1 (HO-1) é uma enzima antioxidante com um papel importante na manutenção da função endotelial, e a sua indução por certas moléculas se mostra potencialmente benéfica frente à característica deletéria destas condições sobre o endotélio. Já foi demonstrado anteriormente que a produção de HO-1 pode ser induzida por iodeto de potássio (KI). Portanto, nós avaliamos os efeitos do KI sobre a citotoxicidade e expressão de HO-1 por células de veia de cordão umbilical humano (HUVECs) após incubação com o plasma de mulheres diagnosticadas com GH ou PE. Métodos Células de veia de cordão umbilical humano foram incubadas com pool de plasma de gestantes saudáveis (n = 12), gestantes com GH (n = 10) ou gestantes com PE (n = 11) com ou sem a adição de KI por 24 horas para avaliar a citotoxicidade através da dosagem de lactato desidrogenase e produção de HO-1 por ELISA. Foi realizada ANOVA seguida de teste de Dunnet para múltiplas comparações entre os grupos estudados, considerando significativos valores de p ≤ 0,05. Resultados A solução de KI (1.000 µM) reduziu a produção de HO-1 no grupo GH (p = 0.0018) e a citotoxicidade no grupo PE (p = 0.0143); o tratamento com KI não afetou a produção de HO-1 por HUVECs incubadas com o plasma do grupo PE. Conclusão Nossos achados sugerem que o KI atenua os efeitos do plasma de gestantes com GH ocasionando a diminuição da produção de HO-1; plasma do grupo PE não induziu a produção de HO-1 em HUVECs em comparação ao grupo saudável, e nossa hipótese é a de que tal achado pode ser devido a mecanismos pós-transcricionais em resposta a uma superestimulação da produção de HO-1 nos estágios iniciais da gravidez.

NLRP3 Activation and Its Relationship to Endothelial Dysfunction and Oxidative Stress: Implications for Preeclampsia and Pharmacological Interventions.

Preeclampsia (PE) is a specific syndrome of human pregnancy, being one of the main causes of maternal death. Persistent inflammation in the endothelium stimulates the secretion of several inflammatory mediators, activating different signaling patterns. One of these mechanisms is related to NLRP3 activation, initiated by high levels of danger signals such as cholesterol, urate, and glucose, producing IL-1, IL-18, and cell death by pyroptosis. Furthermore, reactive oxygen species (ROS), act as an intermediate to activate NLRP3, contributing to subsequent inflammatory cascades and cell damage. Moreover, increased production of ROS may elevate nitric oxide (NO) catabolism and consequently decrease NO bioavailability. NO has many roles in immune responses, including the regulation of signaling cascades. At the site of inflammation, vascular endothelium is crucial in the regulation of systemic inflammation with important implications for homeostasis. In this review, we present the important role of NLRP3 activation in exacerbating oxidative stress and endothelial dysfunction. Considering that the causes related to these processes and inflammation in PE remain a challenge for clinical practice, the use of drugs related to inhibition of the NLRP3 may be a good option for future solutions for this disease.

Different profiles of circulating arginase 2 in subtypes of preeclampsia pregnant women.

BACKGROUND AND AIMS: Preeclampsia (PE) is a gestational hypertensive disease responsible for high maternal and fetal morbidity and mortality. The increase in blood pressure is associated with a decrease in the bioavailability of nitric oxide (NO). Arginase interferes with NO production consuming L-arginine, a substrate required by endothelial NO synthase to NO formation. No previous study has quantified the circulating levels of the two arginase isoforms (arginase 1 and arginase 2) in the plasma of pregnant women with PE. Therefore, our objective is to evaluate these plasma levels in healthy pregnant women and PE with or without severe features and who respond or not to antihypertensive therapy. METHODS: We compared 29 healthy pregnant women with 56 pregnant women with PE, who were also divided into with severe features (n = 24) or without severe features (n = 32) and into responsive (n = 29) or nonresponsive to antihypertensive therapy (n = 27). We quantified the plasmatic expression of arginase 1 and arginase 2 by ELISA kits. RESULTS: While similar levels of arginase 1 were found among groups, lower arginase 2 plasma levels were found in PE without severe features and responsive to antihypertensive drugs when compared to healthy pregnant women. There was no difference between arginase 2 levels in PE with severe features and nonresponsive group when compared to healthy pregnant women. CONCLUSION: This shows different circulation profiles of arginase 2 among groups, suggesting the existence of mechanisms of arginase 2 modulation in pregnant women with PE associated with the severity of the disease and responsiveness to antihypertensive treatment.

Monocytes from preeclamptic women previously treated with silibinin attenuate oxidative stress in human endothelial cells.

Objective: To investigate whether the supernatant from monocytes of preeclamptic and normotensive pregnant women, cultured in vitro with silibinin, can modulate oxidative stress in HUVEC.Methods: Concentrations of IL-1ß, IL-10, and TNF-α in monocyte culture supernatants were determined by ELISA. HUVEC and their supernatant cultures were employed for determination of NO, nitrite and nitrate, lipid peroxidation, and hemeoxygenase-1 (HO-1).Results: HUVEC treatment with supernatant of preeclamptic monocytes cultured with silibinin produced increased levels of nitrite, reduced lipid peroxidation, and increased HO-1.Conclusion: Supernatant of monocytes from preeclamptic women induce oxidative stress in HUVEC which can be reduced by silibinin treatment.Abbreviations: DAF-FMTM, Diaminofluorescein-FM; EDTA, Ethylenediaminetetraacetic acid; HO-1, heme oxygenase-1; HPLC, high-performance liquid chromatography; HUVEC, human umbilical vein endothelial cell; MDA, malondialdehyde; NO, nitric oxide; NT, normotensive; PE, preeclampsia; ROS, reactive oxygen species; Sb, silibinin.

Resveratrol and grape juice: Effects on redox status and nitric oxide production of endothelial cells in in vitro preeclampsia model.

Preeclampsia (PE) is a hypertensive disorder of pregnancy and it is one of the main causes of maternal and fetal morbidity and mortality worldwide. It is known that oxidative stress plays a role in its pathophysiology, therefore we investigated the effects of trans-resveratrol, a potent antioxidant, on the Nrf2/ARE pathway, nitric oxide (NO) production, and reactive oxygen species (ROS) levels in an in vitro model of PE. Plasma from PE patients increased ARE activity in endothelial cells compared with plasma from healthy pregnant (HP), and the addition of resveratrol was able to potentiate this increase only in PE. Resveratrol also decreased ROS levels in the cells incubated with plasma from PE. Based on these results, we performed a pilot clinical study to compare the effects of serum from PE women before and 1 h after ingestion of polyphenol-rich whole red grapefruit juice incubated on endothelial cells, since grapefruit contains large amounts of resveratrol. Serum from PE patients, obtained one hour after juice intake, decreased antioxidants markers in cells compared with the serum before juice intake, besides, it increased NO production. In conclusion, resveratrol and polyphenol-rich red grape juice have potentially beneficial effects on endothelial cells incubated with PE plasma/serum, which could aid in the management of PE.

SIRT1-dependent effects of resveratrol and grape juice in an in vitro model of preeclampsia.

Preeclampsia (PE) is a multifactorial hypertensive disorder of pregnancy that is partly responsible for both maternal and fetal morbidity and mortality levels worldwide. It has been recently discovered that sirtuin-1 (SIRT1) is reduced in the circulation and in an in vitro model of PE. Therefore, in this study, we investigated the effects of trans-resveratrol, a potent antioxidant and activator of SIRT1, on oxidative stress and nitric oxide (NO) production in an in vitro model of PE compared to gestational hypertensive (GH) and healthy pregnant (HP) women. Furthermore, we also evaluated the effects of an acute intake of grape juice on women with PE to assess whether it could mimic in vitro trans-resveratrol supplementation. (1) In the GH group, resveratrol decreased intracellular reactive oxygen species (ROS) and increased their antioxidant capacity, while inhibiting SIRT1 reestablished previous levels. (2) In PE, inhibition of SIRT1 increased antioxidant activity. (3) Intracellular NO and supernatant nitrite levels were increased by inhibiting SIRT1 in the PE group. (4) Grape juice intake increased intracellular NO levels versus before grape juice intake control; however, the inhibition of SIRT1 before grape juice intake initially increased NO, but decreased it 1 h after grape juice intake. In conclusion, activating SIRT1 by using resveratrol alone may not be beneficial to women with PE, and GH and PE seem to have different responsive mechanisms to this molecule. Furthermore, grape juice intake seems to have different effects compared to resveratrol supplementation alone in this in vitro model of PE, demonstrating the potential of the combination of other biologically active molecules from grape juice over the SIRT1-eNOS-NO in PE treatment.

Interaction Between NOS3 and HMOX1 on Antihypertensive Drug Responsiveness in Preeclampsia.

OBJECTIVE: We examined the interaction of polymorphisms in the genes heme oxygenase-1 (HMOX1) and nitric oxide synthase (NOS3) in patients with preeclampsia (PE) as well as the responsiveness to methyldopa and to total antihypertensive therapy. METHODS: The genes HMOX1 (rs2071746, A/T) and NOS3 (rs1799983, G/T) were genotyped using TaqMan allele discrimination assays (Applied Biosystems, Foster City, CA, USA ), and the levels of enzyme heme oxygenase-1 (HO-1) were measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: We found interactions between genotypes of the HMOX-1 and NOS3 genes and responsiveness to methyldopa and that PE genotyped as AT presents lower levels of protein HO-1 compared with AA. CONCLUSION: We found interactions between the HMOX-1 and NOS3 genes and responsiveness to methyldopa and that the HMOX1 polymorphism affects the levels of enzyme HO-1 in responsiveness to methyldopa and to total antihypertensive therapy. These data suggest impact of the combination of these two polymorphisms on antihypertensive responsiveness in PE.

OBJETIVO: Examinamos a interação dos polimorfismos nos genes heme oxigenase-1 (HMOX1) eóxido nítrico sintase (NOS3) em pacientes com pré-eclâmpsia (PE)bem como as capacidades de resposta à metildopa e à terapia anti-hipertensiva. MéTODOS: Os polimorfismos nos genes HMOX1 (rs2071746, A/T) e NOS3 (rs1799983, G/T) foram genotipados usando TaqMan allele discrimination assays (Applied Biosystems, Foster City, CA, EUA), e os níveis da enzima heme oxigenase-1 (HO-1) foram medidos por enzyme-linked immunosorbent assay (ELISA). RESULTADOS: Foram encontradas interações entre os genótipos da HMOX-1 e NOS3 e responsividade à metildopa, e que pacientes genotipados como AT apresentam níveis mais baixos de proteína HO-1 em comparação com o genótipo AA. CONCLUSãO: Foram encontradas interações entre os genes HMOX-1 e NOS3 e responsividade à metildopa e que o polimorfismo localizado no gene HMOX1 afeta os níveis de enzima HO-1 na resposta à metildopa e à terapia anti-hipertensiva. Esses dados sugerem o impacto da combinação desses dois polimorfismos na resposta anti-hipertensiva na PE.