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1.
Hypertension ; 81(7): 1537-1549, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38752345

ABSTRACT

BACKGROUND: Preeclampsia is a multifaceted syndrome that includes maternal vascular dysfunction. We hypothesize that increased placental glycolysis and hypoxia in preeclampsia lead to increased levels of methylglyoxal (MGO), consequently causing vascular dysfunction. METHODS: Plasma samples and placentas were collected from uncomplicated and preeclampsia pregnancies. Uncomplicated placentas and trophoblast cells (BeWo) were exposed to hypoxia. The reactive dicarbonyl MGO and advanced glycation end products (Nε-(carboxymethyl)lysine [CML], Nε-(carboxyethyl)lysine [CEL], and MGO-derived hydroimidazolone [MG-H]) were quantified using liquid chromatography-tandem mass spectrometry. The activity of GLO1 (glyoxalase-1), that is, the enzyme detoxifying MGO, was measured. The impact of MGO on vascular function was evaluated using wire/pressure myography. The therapeutic potential of the MGO-quencher quercetin and mitochondrial-specific antioxidant mitoquinone mesylate (MitoQ) was explored. RESULTS: MGO, CML, CEL, and MG-H2 levels were elevated in preeclampsia-placentas (+36%, +36%, +25%, and +22%, respectively). Reduced GLO1 activity was observed in preeclampsia-placentas (-12%) and hypoxia-exposed placentas (-16%). Hypoxia-induced MGO accumulation in placentas was mitigated by the MGO-quencher quercetin. Trophoblast cells were identified as the primary source of MGO. Reduced GLO1 activity was also observed in hypoxia-exposed BeWo cells (-26%). Maternal plasma concentrations of CML and the MGO-derived MG-H1 increased as early as 12 weeks of gestation (+16% and +17%, respectively). MGO impaired endothelial barrier function, an effect mitigated by MitoQ, and heightened vascular responsiveness to thromboxane A2. CONCLUSIONS: This study reveals the accumulation of placental MGO in preeclampsia and upon exposure to hypoxia, demonstrates how MGO can contribute to vascular impairment, and highlights plasma CML and MG-H1 levels as promising early biomarkers for preeclampsia.


Subject(s)
Biomarkers , Placenta , Pre-Eclampsia , Pyruvaldehyde , Pre-Eclampsia/metabolism , Pre-Eclampsia/physiopathology , Pre-Eclampsia/blood , Humans , Female , Pyruvaldehyde/metabolism , Pyruvaldehyde/blood , Pregnancy , Placenta/metabolism , Biomarkers/metabolism , Biomarkers/blood , Adult , Glycation End Products, Advanced/metabolism , Trophoblasts/metabolism , Lactoylglutathione Lyase/metabolism
2.
Reprod Sci ; 28(8): 2186-2199, 2021 08.
Article in English | MEDLINE | ID: mdl-33523425

ABSTRACT

Preeclampsia complicates 5-8% of all pregnancies worldwide, and although its pathophysiology remains obscure, placental oxidative stress and mitochondrial abnormalities are considered to play a key role. Mitochondrial abnormalities in preeclamptic placentae have been described, but the extent to which mitochondrial content and the molecular pathways controlling this (mitochondrial biogenesis and mitophagy) are affected in preeclamptic placentae is unknown. Therefore, in preeclamptic (n = 12) and control (n = 11) placentae, we comprehensively assessed multiple indices of placental antioxidant status, mitochondrial content, mitochondrial biogenesis, mitophagy, and mitochondrial fusion and fission. In addition, we also explored gene expression profiles related to inflammation and apoptosis. Preeclamptic placentae were characterized by higher levels of oxidized glutathione, a higher total antioxidant capacity, and higher mRNA levels of the mitochondrial-located antioxidant enzyme manganese-dependent superoxide dismutase 2 compared to controls. Furthermore, mitochondrial content was significantly lower in preeclamptic placentae, which was accompanied by an increased abundance of key constituents of glycolysis. Moreover, mRNA and protein levels of key molecules involved in the regulation of mitochondrial biogenesis were lower in preeclamptic placentae, while the abundance of constituents of the mitophagy, autophagy, and mitochondrial fission machinery was higher compared to controls. In addition, we found evidence for activation of apoptosis and inflammation in preeclamptic placentae. This study is the first to comprehensively demonstrate abnormalities at the level of the mitochondrion and the molecular pathways controlling mitochondrial content/function in preeclamptic placentae. These aberrations may well contribute to the pathophysiology of preeclampsia by upregulating placental inflammation, oxidative stress, and apoptosis. Graphical Abstract.


Subject(s)
Mitochondria/metabolism , Oxidative Stress/physiology , Placenta/metabolism , Pre-Eclampsia/metabolism , Adult , Antioxidants/metabolism , Apoptosis/physiology , Female , Humans , Inflammation/metabolism , Pregnancy , Superoxide Dismutase/metabolism , Trophoblasts/metabolism
3.
PLoS One ; 16(1): e0245155, 2021.
Article in English | MEDLINE | ID: mdl-33434211

ABSTRACT

INTRODUCTION: Impaired utero-placental perfusion is a well-known feature of early preeclampsia and is associated with placental hypoxia and oxidative stress. Although aberrations at the level of the mitochondrion have been implicated in PE pathophysiology, whether or not hypoxia-induced mitochondrial abnormalities contribute to placental oxidative stress is unknown. METHODS: We explored whether abnormalities in mitochondrial metabolism contribute to hypoxia-induced placental oxidative stress by using both healthy term placentae as well as a trophoblast cell line (BeWo cells) exposed to hypoxia. Furthermore, we explored the therapeutic potential of the antioxidants MitoQ and quercetin in preventing hypoxia-induced placental oxidative stress. RESULTS: Both in placental explants as well as BeWo cells, hypoxia resulted in reductions in mitochondrial content, decreased abundance of key molecules involved in the electron transport chain and increased expression and activity of glycolytic enzymes. Furthermore, expression levels of key regulators of mitochondrial biogenesis were decreased while the abundance of constituents of the mitophagy, autophagy and mitochondrial fission machinery was increased in response to hypoxia. In addition, placental hypoxia was associated with increased oxidative stress, inflammation, and apoptosis. Moreover, experiments with MitoQ revealed that hypoxia-induced reactive oxygen species originated from the mitochondria in the trophoblasts. DISCUSSION: This study is the first to demonstrate that placental hypoxia is associated with mitochondrial-generated reactive oxygen species and significant alterations in the molecular pathways controlling mitochondrial content and function. Furthermore, our data indicate that targeting mitochondrial oxidative stress may have therapeutic benefit in the management of pathologies related to placental hypoxia.


Subject(s)
Mitochondria/metabolism , Organelle Biogenesis , Oxidative Stress , Pre-Eclampsia/metabolism , Reactive Oxygen Species/metabolism , Trophoblasts/metabolism , Cell Hypoxia , Cell Line , Female , Humans , Mitochondria/pathology , Pre-Eclampsia/pathology , Pregnancy , Trophoblasts/pathology
4.
PLoS Biol ; 18(11): e3000885, 2020 11.
Article in English | MEDLINE | ID: mdl-33170835

ABSTRACT

Hypertension is the most important cause of death and disability in the elderly. In 9 out of 10 cases, the molecular cause, however, is unknown. One mechanistic hypothesis involves impaired endothelium-dependent vasodilation through reactive oxygen species (ROS) formation. Indeed, ROS forming NADPH oxidase (Nox) genes associate with hypertension, yet target validation has been negative. We re-investigate this association by molecular network analysis and identify NOX5, not present in rodents, as a sole neighbor to human vasodilatory endothelial nitric oxide (NO) signaling. In hypertensive patients, endothelial microparticles indeed contained higher levels of NOX5-but not NOX1, NOX2, or NOX4-with a bimodal distribution correlating with disease severity. Mechanistically, mice expressing human Nox5 in endothelial cells developed-upon aging-severe systolic hypertension and impaired endothelium-dependent vasodilation due to uncoupled NO synthase (NOS). We conclude that NOX5-induced uncoupling of endothelial NOS is a causal mechanism and theragnostic target of an age-related hypertension endotype. Nox5 knock-in (KI) mice represent the first mechanism-based animal model of hypertension.


Subject(s)
Hypertension/physiopathology , NADPH Oxidase 5/genetics , Nitric Oxide/metabolism , Adult , Age Factors , Aged , Animals , Endothelial Cells , Endothelium, Vascular , Female , Gene Knock-In Techniques/methods , Humans , Hypertension/genetics , Hypertension/metabolism , Male , Membrane Proteins/genetics , Mice , Middle Aged , NADPH Oxidase 5/metabolism , NADPH Oxidases/genetics , NADPH Oxidases/metabolism , Nitric Oxide/genetics , Nitric Oxide Synthase/genetics , Nitric Oxide Synthase/metabolism , Nitric Oxide Synthase Type III/genetics , Nitric Oxide Synthase Type III/metabolism , Reactive Oxygen Species
5.
Hypertens Res ; 43(12): 1361-1374, 2020 12.
Article in English | MEDLINE | ID: mdl-32733105

ABSTRACT

Preeclampsia (PE) is a pregnancy-related disorder characterized by hypertension and proteinuria that affects 3-10% of all pregnancies. Although its pathophysiology remains obscure, placental hypoxia-induced oxidative stress and alterations in vascular function, morphology, and endothelial barrier integrity are considered to play a key role in the development of preeclampsia. In this study, placental villous explants of noncomplicated placentae and BeWo cells were subjected to hypoxia. The effect of placental hypoxic-conditioned medium (HCM) on intraluminal-induced contraction and endothelial barrier integrity in chorionic arteries was investigated using pressure myography. The impact of BeWo cell HCM on endothelial cell viability, reactive oxygen species formation and inflammation was also determined. Alterations in arterial morphology and contractile responsiveness to the thromboxane A2 analog (U46619) after exposure to placental HCM were examined immunohistochemically and by wire myography, respectively. Intraluminal administration of placental HCM induced vasoconstriction and increased the endothelial permeability for KCl, which was concentration-dependently prevented by quercetin. Placental and BeWo cell HCMs decreased endothelial cell viability, increased the production of reactive oxygen species and enhanced the secretion of IL-6 and IL-8. The cross-sectional area of the arterial media was increased upon exposure to placental HCM, which was associated with increased vascular proliferation and contractile responsiveness to U46619, and all of these effects were prevented by the antioxidants quercetin and RRR-α-tocopherol. This study is the first to comprehensively demonstrate the link between factors secreted by placental cells in response to hypoxia and vascular abnormalities and paves the way for new diagnostic approaches and therapies to better protect the maternal vasculature during and after a preeclampsia-complicated pregnancy.


Subject(s)
Endothelium, Vascular/physiopathology , Hypoxia/physiopathology , Placenta/physiopathology , 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid , Cell Line , Culture Media, Conditioned , Endothelial Cells/physiology , Endothelium, Vascular/metabolism , Female , Humans , In Vitro Techniques , Permeability , Placenta/blood supply , Pre-Eclampsia/etiology , Pregnancy , Reactive Oxygen Species/metabolism , Vasoconstriction
6.
PLoS One ; 13(8): e0202648, 2018.
Article in English | MEDLINE | ID: mdl-30142162

ABSTRACT

INTRODUCTION: Preeclampsia is a major health problem in human pregnancy, severely complicating 5-8% of all pregnancies. The emerging molecular mechanism is that conditions like hypoxic stress trigger the release of placental messengers into the maternal circulation, which causes preeclampsia. Our objective was to develop an in vitro model, which can be used to further elucidate the molecular mechanisms of preeclampsia and which might be used to find a remedy. METHODS: Human non-complicated term placentas were collected. Placental explants were subjected to severe hypoxia and the conditioned media were added to chorionic arteries that were mounted into a myograph. Contractile responses of the conditioned media were determined, as well as effects on thromboxane-A2 (U46619) induced contractility. To identify the vasoactive compounds present in the conditioned media, specific receptor antagonists were evaluated. RESULTS: Factors released by placental explants generated under severe hypoxia induced an increased vasoconstriction and vascular contractility to thromboxane-A2. It was found that agonists for the angiotensin-I and endothelin-1 receptor released by placental tissue under severe hypoxia provoke vasoconstriction. The dietary antioxidant quercetin could partially prevent the acute and sustained vascular effects in a concentration-dependent manner. DISCUSSION: Both the acute vasoconstriction, as well as the increased contractility to U46619 are in line with the clinical vascular complications observed in preeclampsia. Data obtained with quercetin supports that our model opens avenues for e.g. nutritional interventions aimed at treating or preventing preeclampsia.


Subject(s)
Constriction, Pathologic/genetics , Placenta/metabolism , Pre-Eclampsia/genetics , Vasoconstriction/genetics , 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid/pharmacology , Cell Hypoxia/drug effects , Cell Hypoxia/genetics , Chorion/blood supply , Chorion/metabolism , Chorion/pathology , Constriction, Pathologic/physiopathology , Culture Media, Conditioned/pharmacology , Endothelium, Vascular/drug effects , Endothelium, Vascular/pathology , Female , Humans , Hypoxia/genetics , Hypoxia/pathology , Muscle Contraction/drug effects , Myography , Placenta/pathology , Pre-Eclampsia/metabolism , Pre-Eclampsia/physiopathology , Pregnancy , Vasoconstriction/drug effects , Vasoconstrictor Agents/pharmacology
7.
Curr Neurovasc Res ; 14(4): 306-315, 2017.
Article in English | MEDLINE | ID: mdl-28982333

ABSTRACT

BACKGROUND: The regulation of cerebral arterial vasomotor tone involves several mechanisms. The role of sympathetic nerves and the adrenergic neurotransmitter, noradrenaline (NA), has been the subject of debate for decades. Moreover, the specific role of endothelin-1 (ET-1) in cerebral arterial vasoconstriction has not been elucidated to date. In this study, we evaluated the contribution of NA and ET-1 to cerebral artery vasoconstriction. METHODS: Arterial responses of rat middle cerebral arteries, and human pial cerebral arteries to cumulative concentrations of NA and ET-1, and to Electrical Field Stimulation (EFS), were evaluated. To assess the role of NA and ET-1 when EFS was applied, experiments were performed in the presence of adrenergic, neurogenic, and endothelin-1 receptor modulators. RESULTS: We found that vasoconstriction of cerebral arteries following EFS requires the application of exogenous NA, whereas neither EFS nor NA alone induced vasoconstriction. The observed vasoconstriction was abolished by α-adrenoreceptor antagonist, catecholamine-release inhibitor, blockade of the perivascular neurons, and by the endothelin-2 receptor antagonist (BQ123). CONCLUSION: Based on our results, cerebral artery vasoconstriction requires simultaneous neurogenic and adrenergic activation and is ET-1 dependent. We hypothesize that NA modulates the release of ET-1. Upon release, ET-1 binds to the ETA-receptor on smooth muscle cells inducing cerebral artery vasoconstriction.


Subject(s)
Cerebral Arteries/physiology , Endothelin-1/pharmacology , Norepinephrine/pharmacology , Receptor Cross-Talk/physiology , Receptors, Endothelin/physiology , Vasoconstriction/physiology , Adult , Aged , Animals , Cerebral Arteries/drug effects , Dose-Response Relationship, Drug , Female , Humans , Male , Middle Aged , Rats , Rats, Inbred WKY , Receptor Cross-Talk/drug effects , Receptors, Endothelin/agonists , Vasoconstriction/drug effects , Young Adult
8.
Hypertens Res ; 40(8): 738-745, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28298655

ABSTRACT

Endothelin 1 (ET-1), a potent vasoconstrictor, pro-mitogenic and pro-inflammatory peptide, may promote development of endothelial dysfunction and arterial remodeling. ET-1 can be formed through cleavage of big-ET-1 by endothelin-converting enzyme (ECE) or neutral endopeptidase (NEP). We investigated whether chronic treatment with the novel dual NEP/ECE inhibitor SOL1 improves functional and structural properties of resistance-sized arteries of 32-week-old male spontaneously hypertensive rats (SHR). SHR received a chronic 4-week treatment with SOL1, losartan or hydralazine. We then compared effects of inhibition of NO synthase (NOS) (100 µM l-NAME), blockade of ETA- and ETB-receptors (10 µM bosentan) and stimulation of the endothelium with 0.001-10 µM acetylcholine (ACh) in isolated third-order mesenteric resistance arteries. Losartan and hydralazine significantly lowered blood pressure. Losartan decreased the media-to-lumen ratio of resistance arteries. l-NAME (1) increased arterial contractile responses to K+ (5.9-40 mM) in the losartan, SOL1 and vehicle group and (2) increased the sensitivity to phenylephrine (PHE; 0.16-20 µM) in the SOL1 group but not in the losartan, hydralazine and vehicle group. Relaxing responses to ACh in the absence or presence of l-NAME during contractions induced by either 10 µM PHE or 40 mM K+ were not altered by any in vivo treatment. Acute treatment with bosentan did, however, significantly improve maximal relaxing responses involving endothelium-derived nitric oxide and -hyperpolarizing factors in the SOL1 group but not in the losartan, hydralazine or vehicle group. Thus, chronic inhibition of NEP/ECE improved basal endothelial function but did not alter blood pressure, resistance artery structure and stimulated endothelium-dependent relaxing responses in 32-week-old SHR.


Subject(s)
Endothelin-Converting Enzymes/antagonists & inhibitors , Endothelium, Vascular/physiopathology , Enzyme Inhibitors/therapeutic use , Hypertension/drug therapy , Hypertension/physiopathology , Mesenteric Arteries/physiopathology , Neprilysin/antagonists & inhibitors , Protease Inhibitors/therapeutic use , Angiotensin II Type 1 Receptor Blockers/therapeutic use , Animals , Benzazepines/therapeutic use , Bosentan , Endothelium, Vascular/drug effects , Hydralazine/therapeutic use , Losartan/therapeutic use , Mesenteric Arteries/drug effects , Muscle Contraction/drug effects , NG-Nitroarginine Methyl Ester/therapeutic use , Nitric Oxide Synthase Type III/antagonists & inhibitors , Rats , Rats, Inbred SHR , Sulfonamides/therapeutic use , Vascular Resistance/drug effects
9.
Biochim Biophys Acta Gen Subj ; 1861(5 Pt A): 1177-1189, 2017 May.
Article in English | MEDLINE | ID: mdl-28286015

ABSTRACT

BACKGROUND: Rutin intake is associated with a reduced risk of cardiovascular disease (CVD). The exact mechanism by which rutin can protect against CVD development is still enigmatic. Since, rutin is a compound with a relatively short half-life, the direct antioxidant effect of rutin cannot explain the long-lasting effect on human health. We hypothesized that rutin next to its direct antioxidant effect that improves endothelial function, may also induce an adaptive response in endogenous antioxidant systems. METHODS AND RESULTS: In Human Umbilical Vein Endothelial Cells (HUVECs), the direct antioxidant effect was confirmed. During scavenging of Reactive Oxygen Species (ROS), rutin is oxidized into a quinone derivative. HUVECs pretreated with rutin quinone became better protected against a second challenge with oxidative stress 3h later. LC-MS/MS analysis indicated that rutin quinone targets cysteine 151 of Keap1. Moreover, we found that the quinone is an inhibitor of the selenoprotein thioredoxin reductase 1. These properties correlated with an activation of Nrf2 and upregulation of Glutamate Cysteine Ligase, the rate-limiting enzyme of glutathione synthesis, while NF-κB and HIF activation became blunted by rutin treatment. Furthermore, rutin was found to prevent hydrogen peroxide from impairing relaxation of human chorionic plate placental vessels, which may help to protect endothelial function. CONCLUSION AND SIGNIFICANCE: Rutin functions as an antioxidant and is oxidized into a quinone that upregulates the Nrf2-mediated endogenous antioxidant response. This mechanism suggests that rutin selectively exerts its protective effects in regions with increased oxidative stress, and explains how rutin reduces the risk of developing CVD. GENERAL SIGNIFICANCE: The newly found mechanism behind the long-term protection of rutin against cardiovascular disease, the selective upregulation of endogenous antioxidant systems, contributes to the further understanding why rutin can reduce the risk on developing cardiovascular disease.


Subject(s)
Adaptation, Physiological/drug effects , Arterioles/drug effects , Human Umbilical Vein Endothelial Cells/drug effects , NF-E2-Related Factor 2/metabolism , Oxidative Stress/drug effects , Protective Agents/pharmacology , Rutin/pharmacology , Antioxidants/pharmacology , Arterioles/metabolism , Cells, Cultured , Female , Glutamate-Cysteine Ligase/metabolism , HEK293 Cells , Half-Life , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Hydrogen Peroxide/pharmacology , NF-kappa B/metabolism , Oxidation-Reduction/drug effects , Placenta/drug effects , Placenta/metabolism , Pregnancy , Reactive Oxygen Species/metabolism , Up-Regulation/drug effects
10.
Eur Respir J ; 46(4): 1084-94, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26113671

ABSTRACT

Limited literature sources implicate mast-cell mediator chymase in the pathologies of pulmonary hypertension and pulmonary fibrosis. However, there is no evidence on the contribution of chymase to the development of pulmonary hypertension associated with lung fibrosis, which is an important medical condition linked with increased mortality of patients who already suffer from a life-threatening interstitial lung disease.The aim of this study was to investigate the role of chymase in this particular pulmonary hypertension form, by using a bleomycin-induced pulmonary hypertension model.Chymase inhibition resulted in attenuation of pulmonary hypertension and pulmonary fibrosis, as evident from improved haemodynamics, decreased right ventricular remodelling/hypertrophy, pulmonary vascular remodelling and lung fibrosis. These beneficial effects were associated with a strong tendency of reduction in mast cell number and activity, and significantly diminished chymase expression levels. Mechanistically, chymase inhibition led to attenuation of transforming growth factor ß1 and matrix-metalloproteinase-2 contents in the lungs. Furthermore, chymase inhibition prevented big endothelin-1-induced vasoconstriction of the pulmonary arteries.Therefore, chymase plays a role in the pathogenesis of pulmonary hypertension associated with pulmonary fibrosis and may represent a promising therapeutic target. In addition, this study may provide valuable insights on the contribution of chymase in the pulmonary hypertension context, in general, regardless of the pulmonary hypertension form.


Subject(s)
Chymases/metabolism , Chymases/physiology , Hypertension, Pulmonary/physiopathology , Lung/physiopathology , Pulmonary Fibrosis/physiopathology , Animals , Bleomycin/chemistry , Chymases/antagonists & inhibitors , Disease Models, Animal , Endothelin-1/metabolism , Enzyme-Linked Immunosorbent Assay , Hemodynamics , Humans , Hypertrophy, Right Ventricular/enzymology , Immunohistochemistry , Lung/enzymology , Lung/metabolism , Mast Cells/enzymology , Matrix Metalloproteinase 2/metabolism , Mesocricetus , Pulmonary Artery/metabolism , Pulmonary Artery/physiopathology , Radioimmunoassay , Random Allocation , Transforming Growth Factor beta1/metabolism
11.
Eur J Pharmacol ; 746: 132-7, 2015 Jan 05.
Article in English | MEDLINE | ID: mdl-25449042

ABSTRACT

The food supplement quercetin is used as self-medication for prostate disorders and is known to induce vasorelaxation. The drug tamsulosin is used in the treatment of benign prostatic hyperplasia. A major side effect of tamsulosin is orthostatic hypotension, mediated by vasorelaxation resulting from α1-adrenoceptor blockade. The overlapping profile prompted us to investigate the pharmacodynamic interaction of quercetin with tamsulosin. Since quercetin is extensively metabolized in the intestines and the liver, the metabolites quercetin-3-glucuronide and 4'O-methyl-quercetin were also examined. Vasorelaxation induced by the compounds was tested in rat mesenteric arteries (average diameter: 360±µm) constricted by the α1-adrenoceptor agonist phenylephrine. Tamsulosin (0.1nM) decreased phenylephrine sensitivity 17-fold (n=10). Quercetin (5, 10 and 20µM) also caused a decrease (2-, 4- and 6-fold respectively) of phenylephrine sensitivity, while 10µM of quercetin-3-glucuronide and 4'O-methyl-quercetin decreased this sensitivity (1.5- and 2-fold) only slightly (n=6). The combination of tamsulosin with quercetin or quercetin metabolites proved to be far more potent than the compounds in isolation. The combination of quercetin, quercetin-3-glucuronide or 4'O-methyl-quercetin with tamsulosin decreased the phenylephrine sensitivity approximately 200-, 35- and 150-fold (n=6). The strong pharmacodynamic interaction between the food supplement quercetin and tamsulosin underlines the potential of the impact of supplement-drug interactions that warrant more research.


Subject(s)
Dietary Supplements , Quercetin/pharmacology , Sulfonamides/pharmacology , Vasodilation/drug effects , Animals , Drug Interactions , Hydrogen Peroxide/metabolism , Male , Phenylephrine/pharmacology , Quercetin/metabolism , Rats , Rats, Wistar , Tamsulosin
12.
Life Sci ; 111(1-2): 36-41, 2014 Aug 28.
Article in English | MEDLINE | ID: mdl-25066928

ABSTRACT

AIMS: In engineered cells, endothelin ETA and ETB receptors can heterodimerize. We tested whether this can also be observed in native tissue. MAIN METHODS: Rat mesenteric resistance arteries (rMRA) were maintained in organ culture for 24h to upregulate ETB-mediated contractions in addition to their normal ETA-mediated responses. They were then exposed to 100 nM linear ET-1 (ETB-agonist) labeled with Oregon Green 488 (OG488/L.-ET-1) and/or to 16nM intact ET-1 (ETA/ETB-agonist) labeled with the rhodamine dye TAMRA (TAMRA/ET-1). Two photon laser scanning microscopy (TPLSM) was used for the visualization of their binding in the tissue. Fluorescence Lifetime Imaging Microscopy (FLIM) was employed for measurements of the OG488/L.-ET-1 lifetime in the absence and presence of TAMRA/ET-1. KEY FINDINGS: After incubation with the labeled ligands, medial smooth muscle cells (SMCs) were efficiently stained and became visible under TPLSM. TAMRA/ET-1 bound to all SMCs whereas OG488/L.-ET-1 stained only groups of SMCs. Interaction of the two receptor subtypes in SMC was investigated in double staining experiments. Fluorescence lifetime of OG488/L.-ET-1 was reduced in the presence of TAMRA/ET-1, which indicates the occurrence of Fluorescence Resonant Energy Transfer (FRET) and suggests close proximity of the two receptor subtypes within the arterial wall. SIGNIFICANCE: The methodology that is introduced by these new observations may be useful to assess ET-receptor heterodimerization in biopsies from relevant experimental animal models and human patients.


Subject(s)
Mesenteric Arteries/physiology , Protein Multimerization/physiology , Receptor, Endothelin A/metabolism , Receptor, Endothelin B/metabolism , Animals , Carboxylic Acids , Endothelin-1/metabolism , Male , Microscopy, Confocal , Microscopy, Fluorescence , Organ Culture Techniques , Rats , Rats, Inbred WKY , Rhodamines
13.
J Hypertens ; 30(9): 1799-808, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22828083

ABSTRACT

BACKGROUND: Endothelin-1 (ET1) is a potent vasoconstrictor peptide with pro-mitogenic and pro-inflammatory properties and is therefore of interest in the development of endothelial dysfunction, endothelium-dependent flow-related remodeling, and hypertension-related remodeling. ET1 can be formed through cleavage of big ET1 by endothelin-converting enzyme (ECE) and neutral endopeptidase (NEP). METHOD: We investigated whether the dual NEP/ECE inhibitor SOL1 improves resistance artery function and structure in 12 weeks old spontaneously hypertensive rats (SHRs) and whether arterial structural responses to decreased (-90%) or increased (+100%) blood flow are impaired in young SHRs. To this end two groups of SHRs received chronic 4-week treatment at two different time points (4-8 and 8-12 weeks) prior to the experiment. We compared in-vitro effects of cyclo-oxygenase inhibition (1 µmol/l indomethacine), nitric oxide synthase inhibition (100 µmol/l N(ω)-L-nitro arginine methyl ester), and stimulation of the endothelium by 0.001-10 µmol/l acetylcholine (ACh) in isolated third-order mesenteric arteries of SHRs and aged-matched Wistar-Kyoto (WKY) rats. RESULTS: SOL1 had no effect on blood pressure in SHRs or WKY rats. ACh caused biphasic effects in mesenteric arteries of SHRs. The contractile component (endothelium-derived contractile factor) was absent in WKY and abolished by acute indomethacin administration or chronic SOL1 treatment. Endothelium-derived nitric oxide-type responses did not differ in both strains and were not influenced by SOL1 treatment. Endothelium-derived hyperpolarizing factor-type responses were severely impaired in SHRs as compared to WKY rats and were normalized by chronic SOL1 treatment. In first-order mesenteric arteries, outward flow-induced remodeling was impaired in SHRs. Chronic SOL1 treatment did not restore this response. CONCLUSION: Thus chronic SOL1 treatment during the development of hypertension in SHRs has no effect on blood pressure but improves several aspects of endothelium-dependent vasomotor responses but not arterial remodeling.


Subject(s)
Angiotensin-Converting Enzyme Inhibitors/pharmacology , Endothelium, Vascular/physiology , Mesenteric Arteries/physiopathology , Peptide Hydrolases/drug effects , Protease Inhibitors/pharmacology , Animals , Hypertension/enzymology , Hypertension/physiopathology , Rats , Rats, Inbred SHR , Rats, Inbred WKY
14.
Hypertens Res ; 35(11): 1093-101, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22786567

ABSTRACT

Arteries from young healthy animals respond to chronic changes in blood flow and blood pressure by structural remodeling. We tested whether the ability to respond to decreased (-90%) or increased (+100%) blood flow is impaired during the development of deoxycorticosterone acetate (DOCA)-salt hypertension in rats, a model for an upregulated endothelin-1 system. Mesenteric small arteries (MrA) were exposed to low blood flow (LF) or high blood flow (HF) for 4 or 7 weeks. The bioavailability of vasoactive peptides was modified by chronic treatment of the rats with the dual neutral endopeptidase (NEP)/endothelin-converting enzyme (ECE) inhibitor SOL1. After 3 or 6 weeks of hypertension, the MrA showed hypertrophic arterial remodeling (3 weeks: media cross-sectional area (mCSA): 10±1 × 10(3) to 17±2 × 10(3) µm(2); 6 weeks: 13±2 × 10(3) to 24±3 × 10(3) µm(2)). After 3, but not 6, weeks of hypertension, the arterial diameter was increased (Ø: 385±13 to 463±14 µm). SOL1 reduced hypertrophy after 3 weeks of hypertension (mCSA: 6 × 10(3)±1 × 10(3) µm(2)). The diameter of the HF arteries of normotensive rats increased (Ø: 463±22 µm) but no expansion occurred in the HF arteries of hypertensive rats (Ø: 471±16 µm). MrA from SOL1-treated hypertensive rats did show a significant diameter increase (Ø: 419±13 to 475±16 µm). Arteries exposed to LF showed inward remodeling in normotensive and hypertensive rats (mean Ø between 235 and 290 µm), and infiltration of monocyte/macrophages. SOL1 treatment did not affect the arterial diameter of LF arteries but reduced the infiltration of monocyte/macrophages. We show for the first time that flow-induced remodeling is impaired during the development of DOCA-salt hypertension and that this can be prevented by chronic NEP/ECE inhibition.


Subject(s)
Blood Pressure/physiology , Desoxycorticosterone/adverse effects , Hypertension/pathology , Hypertension/physiopathology , Mesenteric Arteries/pathology , Mesenteric Arteries/physiopathology , Regional Blood Flow/physiology , Animals , Aspartic Acid Endopeptidases/antagonists & inhibitors , Aspartic Acid Endopeptidases/drug effects , Benzazepines/pharmacology , Blood Pressure/drug effects , Cell Movement/drug effects , Cell Movement/physiology , Disease Models, Animal , Endothelin-Converting Enzymes , Enzyme Inhibitors/pharmacology , Hypertension/chemically induced , Hypertrophy/chemically induced , Macrophages/pathology , Metalloendopeptidases/antagonists & inhibitors , Metalloendopeptidases/drug effects , Monocytes/pathology , Neprilysin/antagonists & inhibitors , Neprilysin/drug effects , Rats , Rats, Wistar , Regional Blood Flow/drug effects , Vascular Resistance/drug effects , Vascular Resistance/physiology
15.
Cell Mol Life Sci ; 69(14): 2327-43, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22648375

ABSTRACT

Reactive oxygen species (ROS) are cellular signals but also disease triggers; their relative excess (oxidative stress) or shortage (reductive stress) compared to reducing equivalents are potentially deleterious. This may explain why antioxidants fail to combat diseases that correlate with oxidative stress. Instead, targeting of disease-relevant enzymatic ROS sources that leaves physiological ROS signaling unaffected may be more beneficial. NADPH oxidases are the only known enzyme family with the sole function to produce ROS. Of the catalytic NADPH oxidase subunits (NOX), NOX4 is the most widely distributed isoform. We provide here a critical review of the currently available experimental tools to assess the role of NOX and especially NOX4, i.e. knock-out mice, siRNAs, antibodies, and pharmacological inhibitors. We then focus on the characterization of the small molecule NADPH oxidase inhibitor, VAS2870, in vitro and in vivo, its specificity, selectivity, and possible mechanism of action. Finally, we discuss the validation of NOX4 as a potential therapeutic target for indications including stroke, heart failure, and fibrosis.


Subject(s)
NADPH Oxidases/metabolism , Animals , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Humans , Models, Animal , NADPH Oxidases/antagonists & inhibitors , NADPH Oxidases/genetics , Oxidative Stress/drug effects , Protein Isoforms/antagonists & inhibitors , Protein Isoforms/genetics , Protein Isoforms/metabolism , RNA Interference , RNA, Small Interfering/metabolism , Reactive Oxygen Species/metabolism
16.
J Renin Angiotensin Aldosterone Syst ; 12(3): 208-14, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21163865

ABSTRACT

INTRODUCTION: This study investigated whether variation in the genes encoding for ACE, AGT and AGTR1 modifies the risk of myocardial infarction (MI) related to ACE inhibitors and AT II antagonists. METHODS: A nested case-control study among users of antihypertensive drugs, in whom the polymorphisms ACE-G4656C, ACE-T3892C, AGT-C235T and AGTR1-A1166C were genotyped. RESULTS: Among 613 cases and 3630 controls, the risk of MI was significantly lower among users of ACE inhibitors compared with that in users of other antihypertensives (adjusted OR, 0.78; 95% CI, 0.63-0.97). In patients using ACE inhibitors the largest risk reduction was found in patients carrying the ACE-4656-G allele (GC and GG genotypes) compared with patients carrying the CC genotype (OR, 0.68; 95% CI, 0.53-0.86 and OR, 1.26, 95% CI, 0.78-2.02, respectively). The synergy index for this interaction was statistically significant (SI, 0.58; 95% CI, 0.35-0.95). The risk of MI was reduced in those who were current users of ACE inhibitors those who had been prescribed dosages lower than the equivalent of 1 defined daily dose (DDD) and those having the AGTR1-1166AC or AA genotype compared with that in users of ACE inhibitors with the AGTR1-1166CC genotype (SI, 3.67; 95% CI,1.18-11.4). None of the polymorphisms modified the effectiveness of AT II antagonists regarding the risk of MI. CONCLUSION: This study shows an interaction between the use of ACE inhibitors and ACE-G4656C polymorphism, and in low doses also with AGTR1-A1166C polymorphism, in the prevention of MI.


Subject(s)
Angiotensin II Type 1 Receptor Blockers/therapeutic use , Angiotensin-Converting Enzyme Inhibitors/therapeutic use , Genetic Predisposition to Disease , Myocardial Infarction/drug therapy , Myocardial Infarction/genetics , Polymorphism, Single Nucleotide/genetics , Renin-Angiotensin System/genetics , Case-Control Studies , Female , Humans , Male , Middle Aged , Risk Factors
17.
Circulation ; 122(3): 273-81, 2010 Jul 20.
Article in English | MEDLINE | ID: mdl-20606119

ABSTRACT

BACKGROUND: Control of peripheral resistance arteries by autonomic nerves is essential for the regulation of blood flow. The signals responsible for the maintenance of vascular neuroeffector mechanisms in the adult, however, remain largely unknown. METHODS AND RESULTS: Here, we report that VEGF( partial differential/ partial differential) mice with low vascular endothelial growth factor (VEGF) levels suffer defects in the regulation of resistance arteries. These defects are due to dysfunction and structural remodeling of the neuroeffector junction, the equivalent of a synapse between autonomic nerve endings and vascular smooth muscle cells, and to an impaired contractile smooth muscle cell phenotype. Notably, short-term delivery of a VEGF inhibitor to healthy mice also resulted in functional and structural defects of neuroeffector junctions. CONCLUSIONS: These findings uncover a novel role for VEGF in the maintenance of arterial neuroeffector function and may help us better understand how VEGF inhibitors cause vascular regulation defects in cancer patients.


Subject(s)
Autonomic Nervous System Diseases/physiopathology , Cardiovascular Diseases/physiopathology , Vascular Endothelial Growth Factor A/genetics , Vascular Resistance/physiology , Vasoconstriction/physiology , Animals , Autonomic Nervous System Diseases/genetics , Cardiovascular Diseases/genetics , Carotid Artery, Common/innervation , Carotid Artery, Common/physiology , Gene Expression/physiology , Gene Transfer Techniques , Lac Operon , Mesenteric Arteries/innervation , Mesenteric Arteries/physiology , Mice , Mice, Transgenic , Muscle, Smooth, Vascular/physiology , Signal Transduction/physiology , Vascular Endothelial Growth Factor A/metabolism , Vascular Endothelial Growth Factor Receptor-1/genetics , Vascular Endothelial Growth Factor Receptor-1/metabolism , Vascular Endothelial Growth Factor Receptor-2/genetics , Vascular Endothelial Growth Factor Receptor-2/metabolism
18.
Front Biosci (Elite Ed) ; 2(3): 1164-8, 2010 06 01.
Article in English | MEDLINE | ID: mdl-20515788

ABSTRACT

Oxidative stress is a key factor driving the aging of cells and arteries. Studies suggest that white blood cell (WBC) telomere length is an index of systemic aging. We, therefore, investigated the association between WBC telomere length and oxidized-LDL, and vascular aging, expressed by the distensibility of the carotid artery. We studied a random population sample of 216 non-smokers and 89, smokers. In all subjects, age and gender- adjusted telomere length was inversely correlated with plasma oxidized-LDL (regression coefficient = -0.656 kb/mg/dL; p=0.0006). Independent of gender, age and mean blood pressure, carotid distensibility increased with telomere length (2.33+/- 1.18 10-3/kPa/kb; p=0.05) but decreased with higher plasma levels of oxidized LDL (-10.7+/- 3.91 10-3/kPa/ mg/dL; p=0.006). Adjusted for gender and age, smokers' telomere length was shorter (6.72 vs 6.91 kb; p=0.014) and plasma oxidized-LDL level higher (0.52 vs 0.46 mg/dL; p=0.03) than in non-smokers. Higher level of oxidized-LDL, is associated with shorter WBC telomeres and increased stiffness of the carotid artery. Smoking is marked by increased oxidative stress in concert with shortened WBC telomere length.


Subject(s)
Carotid Arteries/pathology , Lipoproteins, LDL/blood , Smoking/genetics , Telomere , Adult , Cohort Studies , Female , Humans , Male , Middle Aged
19.
J Renin Angiotensin Aldosterone Syst ; 10(2): 101-8, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19502258

ABSTRACT

INTRODUCTION: We assessed the influence of genetic polymorphisms in the renin-angiotensin system on the risk of diabetes associated with the use of angiotensin II receptor blockers and angiotensin-converting enzyme (ACE) inhibitors. MATERIALS AND METHODS: We performed a matched case-control study among antihypertensive drug users. Pharmacy records and questionnaires were used to ascertain incident diabetes (cases), antihypertensive drug use, and risk factors. Controls did not (yet) have diabetes.We genotyped ACE (G4656C, which is in complete linkage disequilibrium with the ACE insertion/deletion polymorphism), angiotensinogen (M235T), and angiotensin II type 1 receptor (A1166C). RESULTS: Among 495 cases of incident diabetes and 2,624 controls, homozygous 1166C carriers of angiotensin II type 1 receptor who used angiotensin II receptor blockers had an increased risk of diabetes compared to 1166A carriers (interaction odds ratio 5.3 [95% confidence interval: 1.8-16.1]). Homozygous ACE GG subjects who used ACE inhibitors >or= 1 defined daily dose/day had a higher risk of diabetes compared to subjects with the ACE C allele (interaction odds ratio 2.3 [95% confidence interval: 1.2-4.5]). CONCLUSIONS: Angiotensin II receptor blockers increase the occurrence of diabetes in homozygous 1166C carriers of angiotensin II type 1 receptor, but not in 1166A carriers. ACE inhibitors at doses >or= 1 defined daily dose/day increase the risk of diabetes among homozygous ACE GG carriers, but not in 4656C carriers.


Subject(s)
Angiotensin-Converting Enzyme Inhibitors/therapeutic use , Hypertension/drug therapy , Renin-Angiotensin System/genetics , Aged , Case-Control Studies , Diabetes Mellitus, Type 2/genetics , Female , Humans , Hypertension/genetics , Male , Peptidyl-Dipeptidase A/genetics , Polymorphism, Genetic , Receptor, Angiotensin, Type 1/genetics , Risk
20.
Pharmacoepidemiol Drug Saf ; 18(8): 665-71, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19462400

ABSTRACT

PURPOSE: In epidemiological studies, non-response may introduce bias and limit generalizability. In genetic pharmacoepidemiological research, collection of DNA might be a major reason for non-response. We determined reasons for non-response and compared characteristics of non-responders and responders in a pharmacogenetic case-control study. METHODS: Myocardial infarction (MI) cases and controls, who were antihypertensive drug users, were recruited through community pharmacies that participate in the Pharmaco-Morbidity-Record-Linkage-System (PHARMO). The PHARMO database comprises drug dispensing histories of about 2 000 000 subjects from a representative sample of Dutch community pharmacies linked to the national registry of hospital discharges. Independent samples t-test and ANOVA-statistics were used to analyse the differences in continuous variables between responders and non-responders. chi2 statistics and logistic regression were used to compare categorical variables. RESULTS: We approached 1871 cases and 14 102 controls of whom 794 MI cases (42.4%) and 4997 controls (35.4%) responded. We could not approach 2194 patients of whom 63.1% had died and 31.2% moved to another pharmacy. Main reasons for non-response were health problems or hospital stays (16.2%, OR 1.47; 95%CI: 1.00-2.16). Other reasons were old age or dementia (16.9%, OR 1.82; 95%CI: 1.24-2.65). Only a small percentage (1.1%, OR 1.43; 95%CI: 0.41-5.03) mentioned DNA sampling as a reason. About 30% of the non-responders did not give a reason. Women were statistically significantly (p < 0.0005) less willing to participate than men (38.8% versus 31.3%). An association with age was also found (mean age 64.6 versus 66.5 yrs) (p < 0.0005). CONCLUSION: In a pharmacogenetic case-control study fear for genetic screening was not a major reported reason for non-response. Females were less willing to participate than males.


Subject(s)
Antihypertensive Agents/therapeutic use , Myocardial Infarction/drug therapy , Patient Selection , Pharmacogenetics , Research Subjects/psychology , Adult , Aged , Case-Control Studies , Chi-Square Distribution , Correspondence as Topic , Fear , Feasibility Studies , Female , Genetic Privacy , Genetic Testing , Health Knowledge, Attitudes, Practice , Humans , Logistic Models , Male , Middle Aged , Myocardial Infarction/epidemiology , Netherlands/epidemiology , Odds Ratio , Pharmacogenetics/methods , Registries , Risk Assessment , Risk Factors , Surveys and Questionnaires , Telephone
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