Regulation of adrenomedullin release from human endothelial cells by sex steroids and angiotensin-II

It is well documented that there are gender differences in the incidence and patterns of cardiovascular diseases but the reasons are unclear. Sex steroids may modulate the behaviour of vascular endothelial cells, which in turn act by paracrine processes to alter adjacent vascular smooth muscle activity. We hypothesised that the sex steroids alter the percentage of vascular endothelial cells that secrete the vasodilator peptide, adrenomedullin and modify the adrenomedullin-stimulating action of angiotensin-II. The percentage of adrenomedullin-secreting human aortic endothelial cells was determined using the cell immunoblot method. Cells were incubated with selected concentrations of angiotensin-II, oestradiol and testosterone alone and sex steroids in combination with angiotensin-II. Adrenomedullin mRNA expression in endothelial cells was quantified by real-time PCR. It was observed that at 4 h, angiotensin-II increased the percentage of adrenomedullin-secreting cells in a concentration-dependent manner. Testosterone at physiological concentrations was observed to increase the number of adrenomedullin-secreting cells whilst oestradiol had no effect. Addition of testosterone to angiotensin-II resulted in less than additive increases in the number of cells secreting adrenomedullin. Oestradiol reduced the angiotensin-II-induced increase in adrenomedullin-secreting cells. Adrenomedullin mRNA expression was significantly increased by testosterone and there was also a trend for an increase in adrenomedullin mRNA expression, which occurred when cells were incubated with angiotensin-II. Our results point to a complex interplay between the sex steroids and angiotensin-II in regulating adrenomedullin production by human endothelial cells, which may contribute to gender-related differences in vascular disease in humans.

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Citrus bergamiaRisso Elevates Intracellular Ca2+in Human Vascular Endothelial Cells due to Release of Ca2+from Primary Intracellular Stores

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2013/759615 ◽

2013 ◽

Vol 2013 ◽

pp. 1-7 ◽

Cited By ~ 1

Author(s):

Purum Kang ◽

Seung Ho Han ◽

Hea Kyung Moon ◽

Jeong-Min Lee ◽

Hyo-Keun Kim ◽

...

Keyword(s):

Endothelial Cells ◽

Channel Blocker ◽

Vascular Endothelial Cells ◽

Umbilical Vein ◽

Dependent Manner ◽

Vascular Endothelial ◽

Concentration Dependent Manner ◽

Carbonyl Cyanide ◽

Intracellular Ca ◽

Umbilical Vein Endothelial Cells

The purpose of the present study is to examine the effects of essential oil ofCitrus bergamiaRisso (bergamot, BEO) on intracellular Ca2+in human umbilical vein endothelial cells. Fura-2 fluorescence was used to examine changes in intracellular Ca2+concentration[Ca2+]i. In the presence of extracellular Ca2+, BEO increased[Ca2+]i, which was partially inhibited by a nonselective Ca2+channel blocker La3+. In Ca2+-free extracellular solutions, BEO increased[Ca2+]iin a concentration-dependent manner, suggesting that BEO mobilizes intracellular Ca2+. BEO-induced[Ca2+]iincrease was partially inhibited by a Ca2+-induced Ca2+release inhibitor dantrolene, a phospholipase C inhibitor U73122, and an inositol 1,4,5-triphosphate (IP3)-gated Ca2+channel blocker, 2-aminoethoxydiphenyl borane (2-APB). BEO also increased[Ca2+]iin the presence of carbonyl cyanide m-chlorophenylhydrazone, an inhibitor of mitochondrial Ca2+uptake. In addition, store-operated Ca2+entry (SOC) was potentiated by BEO. These results suggest that BEO mobilizes Ca2+from primary intracellular stores via Ca2+-induced and IP3-mediated Ca2+release and affect promotion of Ca2+influx, likely via an SOC mechanism.

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The Study on Angiotensin II Induced-Ferroptosis in Vascular Endothelial Cells

10.21203/rs.3.rs-565583/v1 ◽

2021 ◽

Author(s):

hong fang ◽

Chi liu ◽

Omer Cavdar ◽

Yi Shen

Keyword(s):

Endothelial Cells ◽

Angiotensin Ii ◽

Molecular Marker ◽

Analysis Of Variance ◽

Vascular Endothelial Cells ◽

Dependent Manner ◽

Vascular Endothelial ◽

Ang Ii ◽

Valsartan Group ◽

Dose Dependent

Abstract PurposeTo verify the effect of Angiotensin II on ferroptosis in vascular endothelial cells and clarify the related mechanism. MethodsHUVECs were evaluated for p53, P21,ALOX12, VEGF, MDA,GSH. Molecular marker impact upon AngII-induced ferroptosis was evaluated with students’ t-test，one-way analysis of variance (ANOVA).ResultsAs the concentration of Ang II increased，the level of ALOX12, P53,GSH and MDA increased in HUVECs. The expression of VEGFA in HUVECs is negatively correlated with dose of Ang II. Incubation of HUVECs in AngII and valsartan for 48hr reduces ALOX12, P21, GSH and MDA. Compared with the single AngII group, ALOX12, P21, GSH and MDA in valsartan group was decreased significantly（p=0.000）.In pifithrin-α hydrobromide-treated, ALOX12, P21, GSH and MDA was reduced significantly, as compared to valsartan group（p=0.000）. The most larger reduction in ALOX12, P21,GSH and MDA was pifithrin - α hydrobromide combined with valsartan group. In contrast, the expression of VEGFA increased significantly after HUVECs were treated with pifithrin - α hydrobromide and valsartan（p=0.000）.ConclusionsAngII can induce ferroptosis of vascular endothelial cells in a dose-dependent manner. The mechanism of AngII-induced ferroptosis may be regulated through the signal axis of ATR1,2-p53-ALOX12.

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The Study on Angiotensin II Induced-ferroptosis in Vascular Endothelial Cells

10.21203/rs.3.rs-506444/v1 ◽

2021 ◽

Author(s):

Hong Fang ◽

Chi Liu ◽

Omer Cavdar ◽

Yi Shen

Keyword(s):

Endothelial Cells ◽

Angiotensin Ii ◽

Molecular Marker ◽

Analysis Of Variance ◽

Vascular Endothelial Cells ◽

Dependent Manner ◽

Vascular Endothelial ◽

Ang Ii ◽

Valsartan Group ◽

Dose Dependent

Abstract Background: To verify the effect of Angiotensin II on ferroptosis in vascular endothelial cells and clarify the related mechanism.Methods: HUVECs were evaluated for p53, P21, ALOX12, VEGF, MDA, GSH. Molecular marker impact upon AngII-induced ferroptosis was evaluated with students’ t-test，one-way analysis of variance (ANOVA).Results: As the concentration of Ang II increased，the level of ALOX12, P53, GSH and MDA increased in HUVECs. The expression of VEGFA in HUVECs is negatively correlated with dose of Ang II. Incubation of HUVECs in AngII and valsartan for 48hr reduces ALOX12, P21, GSH and MDA. Compared with the single AngII group, ALOX12, P21, GSH and MDA in valsartan group was decreased significantly（p=0.000）. In pifithrin-α hydrobromide-treated, ALOX12, P21, GSH and MDA was reduced significantly, as compared to valsartan group（p=0.000）. The most larger reduction in ALOX12, P21,GSH and MDA was pifithrin - α hydrobromide combined with valsartan group. In contrast, the expression of VEGFA increased significantly after HUVECs were treated with pifithrin - α hydrobromide and valsartan（p=0.000）.Conclusions: AngII can induce ferroptosis of vascular endothelial cells in a dose-dependent manner. The mechanism of AngII-induced ferroptosis may be regulated through the signal axis of ATR1,2-p53-ALOX12.

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Arsenite stimulates plasma membrane NADPH oxidase in vascular endothelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2001.280.3.l442 ◽

2001 ◽

Vol 280 (3) ◽

pp. L442-L449 ◽

Cited By ~ 89

Author(s):

Karol R. Smith ◽

Linda R. Klei ◽

Aaron Barchowsky

Keyword(s):

Endothelial Cells ◽

Angiotensin Ii ◽

Nadph Oxidase ◽

Oxidase Activity ◽

Vascular Endothelial Cells ◽

Oxidase Inhibitor ◽

Vascular Endothelial ◽

Effect Analysis ◽

Aortic Endothelial Cells ◽

Necrosis Factor

Low-level arsenite treatment of porcine aortic endothelial cells (PAEC) stimulated superoxide accumulation that was attenuated by inhibitors of NAD(P)H oxidase. To demonstrate whether arsenite stimulated NADPH oxidase, intact PAEC were treated with arsenite for up to 2 h and membrane fractions were prepared to measure NADPH oxidase activity. Arsenite (5 μM) stimulated a twofold increase in activity by 1 h, which was inhibited by the oxidase inhibitor diphenyleneiodonium chloride. Direct treatment of isolated membranes with arsenite had no effect. Analysis of NADPH oxidase components revealed that p67phoxlocalized exclusively to membranes of both control and treated cells. In contrast, cytosolic Rac1 translocated to the membrane fractions of cells treated with arsenite or angiotensin II but not with tumor necrosis factor. Immunodepletion of p67phoxblocked oxidase activity stimulated by all three compounds. However, depleting Rac1 inhibited responses only to arsenite and angiotensin II. These data demonstrate that stimulus-specific activation of NADPH oxidase in endothelial cells was the source of reactive oxygen in endothelial cells after noncytotoxic arsenite exposure.

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Modulation of calcium homeostasis in cultured rat aortic endothelial cells by intracellular acidification

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1993.265.4.h1424 ◽

1993 ◽

Vol 265 (4) ◽

pp. H1424-H1433 ◽

Cited By ~ 2

Author(s):

R. C. Ziegelstein ◽

L. Cheng ◽

P. S. Blank ◽

H. A. Spurgeon ◽

E. G. Lakatta ◽

...

Keyword(s):

Endothelial Cells ◽

Vascular Endothelial Cells ◽

Cytosolic Calcium ◽

Ph Sensitive ◽

Vascular Endothelial ◽

Intracellular Acidification ◽

Cytosolic Ph ◽

Aortic Endothelial Cells ◽

Endothelial Monolayers ◽

Aortic Endothelial

Acidosis produces vasodilation in a process that may involve the vascular endothelium. Because synthesis and release of endothelium-derived vasodilatory substances are linked to an increase in cytosolic calcium concentration ([Ca2+]i), we examined the effect of intracellular acidification on cultured rat aortic endothelial cells loaded either with the pH-sensitive probe carboxy-seminaphthorhodafluor-1 or the Ca(2+)-sensitive fluorescent probe indo 1. The basal cytosolic pH (pHi) of endothelial monolayers in a 5% CO2-HCO3- buffer was 7.27 +/- 0.02 and that in a bicarbonate-free solution was 7.22 +/- 0.03. Acidification was induced either by removal of NH4Cl (delta pHi = -0.10 +/- 0.02), changing from a bicarbonate-free to a 5% CO2-HCO3(-)-buffered solution at constant buffer pH (delta pHi = -0.18 +/- 0.03), or changing from a 5% to a 20% CO2-HCO3- solution (delta pHi = -0.27 +/- 0.07). Regardless of the method used, intracellular acidification increased [Ca2+]i as indexed by indo 1 fluorescence. The increase in [Ca2+]i induced by changing from a 5 to a 20% CO2-HCO3- solution was not significantly altered by removal of buffer Ca2+ either before or after depletion of bradykinin- and thapsigargin-sensitive intracellular Ca2+ stores. Thus intracellular acidification of vascular endothelial cells releases Ca2+ into the cytosol either from pH-sensitive intracellular buffer sites, mitochondria, or from bradykinin- and thapsigargin-insensitive intracellular stores. This Ca2+ mobilization may be linked to endothelial synthesis and release of vasodilatory substances during acidosis.

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Biomechanical interactions of Schistosoma mansoni eggs with vascular endothelial cells facilitate egg extravasation

10.1101/2021.09.16.459846 ◽

2021 ◽

Author(s):

Yi-Ting Yeh ◽

Danielle E. Skinner ◽

Ernesto Criado-Hidalgo ◽

Natalie Shee Chen ◽

Antoni Garcia-De Herreros ◽

...

Keyword(s):

Endothelial Cells ◽

Schistosoma Mansoni ◽

Disease Transmission ◽

Vascular Endothelial Cells ◽

Flexible Substrate ◽

Blood Stream ◽

The Body ◽

Dependent Manner ◽

Vascular Endothelial ◽

Maturation State

AbstractThe eggs of the parasitic blood fluke, Schistosoma, are the main drivers of the chronic pathologies associated with schistosomiasis, a disease of poverty afflicting approximately 220 million people worldwide. Eggs laid by Schistosoma mansoni in the bloodstream of the host are encapsulated by vascular endothelial cells (VECs), the first step in the migration of the egg from the blood stream into the lumen of the gut and eventual exit from the body. The biomechanics associated with encapsulation and extravasation of the egg are poorly understood. We demonstrate that S. mansoni eggs induce VECs to form two types of membrane extensions during encapsulation; filopodia that probe eggshell surfaces and intercellular nanotubes that presumably facilitate VEC communication. Encapsulation efficiency, the number of filopodia and intercellular nanotubes, and the length of these structures depend on the egg’s vitality and, to a lesser degree, its maturation state. During encapsulation, live eggs induce VEC contractility and membranous structures formation, in a Rho/ROCK pathway-dependent manner. Using elastic hydrogels embedded with fluorescent microbeads as substrates to culture VECs, live eggs induce VECs to exert significantly greater contractile forces during encapsulation than dead eggs, which leads to 3D deformations on both the VEC monolayer and the flexible substrate underneath. These significant mechanical deformations cause the VEC monolayer tension to fluctuate with eventual rupture of VEC junctions, thus facilitating egg transit out of the blood vessel. Overall, our data on the mechanical interplay between host VECs and the schistosome egg improve our understanding of how this parasite manipulates its immediate environment to maintain disease transmission.

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PGI2 Production In Human Endothelial Cells Cultured In Diabetic And Nondiabetic Serum

10.1055/s-0038-1652118 ◽

1981 ◽

Author(s):

R C Paton ◽

R Guillot ◽

Ph Passa

Keyword(s):

Endothelial Cells ◽

Vascular Endothelial Cells ◽

Vascular Complications ◽

Proliferative Retinopathy ◽

Vascular Endothelial ◽

Human Endothelial Cells ◽

Bovine Thrombin ◽

Control Serum ◽

Umbilical Cords ◽

Cells Cultured

Reduced levels of prostaglandin I2 (PGI2) may contribute to the platelet hyper-reactivity and vascular complications found in diabetes mellitus. This study compared PGI2 production (PGI2-like activity and 6-keto-PGF1α levels) by vascular endothelial cells cultured in the presence of serum from 15 diabetics with proliferative retinopathy (5 treated by surgical hypophysectomy) and 15 sex-matched nondiabetic controls. Endothelial cells from human umbilical veins were cultured in M199 with either 20 % diabetic or control serum. At confluence, cultures were washed and stimulated with 0.1 NIH u/ml bovine thrombin. After 2 min incubation, the supernatant was tested for i)PGI2-like activity on ADP- induced platelet aggregation, results expressed as % inhibition and ii) 6-keto-PGF1α by radioimmunoassay, results expressed as nmol/ml. There was a significant correlation between PGI2-like activity and 6-keto-PGF-1α levels (r 0.78, p<0.001). The liberation of PGI2 from endothelial cells from different umbilical cords varied, but both PGI2-like activity (mean± SEM 21.9± 4.8 vs 28.3± 5.1 p<0.05) and 6-keto-PGF-1α (3.15± 0.68 vs 3.95 ±0.91 nmol/ml, p <0.05)were significantly lower in superantant from cells cultured in the presence of diabetic compared to control serum. PGI2 production was not significantly different in cells cultured with serum from hypophysectomised and nonhypophysectomised diabetics.These results suggest that serum from diabetics with proliferative retinopathy contains factors which impair the release or production of PGI2 by endothelial cells and that this effect is not mediated by the pituitary.

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Regulation of endothelin-1 release from human endothelial cells by sex steroids and angiotensin-II

Peptides ◽

10.1016/j.peptides.2008.02.003 ◽

2008 ◽

Vol 29 (6) ◽

pp. 1057-1061 ◽

Cited By ~ 22

Author(s):

Lachlan J. Pearson ◽

Timothy G. Yandle ◽

M. Gary Nicholls ◽

John J. Evans

Keyword(s):

Endothelial Cells ◽

Angiotensin Ii ◽

Sex Steroids ◽

Human Endothelial Cells ◽

Endothelin 1

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Effect of angiotensin-converting-enzyme inhibition on bradykinin metabolism by vascular endothelial cells

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1993.264.5.h1493 ◽

1993 ◽

Vol 264 (5) ◽

pp. H1493-H1497 ◽

Cited By ~ 3

Author(s):

M. Grafe ◽

C. Bossaller ◽

K. Graf ◽

W. Auch-Schwelk ◽

C. R. Baumgarten ◽

...

Keyword(s):

Endothelial Cells ◽

Angiotensin Converting Enzyme ◽

Vascular Endothelial Cells ◽

Ace Inhibition ◽

Angiotensin Converting Enzyme Inhibition ◽

Dependent Manner ◽

Converting Enzyme ◽

Concentration Dependent Manner ◽

Cell Monolayers ◽

Converting Enzyme Inhibition

The degradation of bradykinin by angiotensin-converting-enzyme (ACE) activity in cultured human endothelial cells was studied by direct measurement of bradykinin and by its effect on the release of endothelium-derived relaxing factors. The half-life of exogenous bradykinin (10,000 pg/ml) was calculated from the decay of the bradykinin concentration as 46 +/- 2 min in cell monolayers, 133 +/- 15 min in conditioned medium, and 24 +/- 2 min in homogenates. Most of the bradykinin-degrading activity in cell monolayers could be inhibited in a concentration-dependent manner by the ACE inhibitors lisinopril, ramiprilat, and captopril. Bradykinin-degrading activity was released into the culture medium containing one-fourth of the bradykinin-degrading activity found in the presence of cell monolayers. In cell homogenates higher unspecific bradykinin-degrading activities were present. The functional consequence of bradykinin degradation was demonstrated by the potentiating effect of ramiprilat on the generation of endothelium-derived relaxing factors nitric oxide and prostacyclin from endothelial cells. The study supports the concept of increased vasodilatory effects of bradykinin during ACE inhibition.

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