scholarly journals Omega-3 fatty acids differentially alter the expression of detoxification enzymes and nitric oxide bioavailability in endothelial cells during IL-6 exposure

2021 ◽  
Vol 42 (Supplement_1) ◽  
Author(s):  
S C R Sherratt ◽  
P Libby ◽  
D L Bhatt ◽  
H Dawoud ◽  
T Malinski ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Umbilical Vein ◽  
Calcium Ionophore ◽  
Detoxification Enzymes ◽  
Omega 3 ◽  
Private Company ◽  
Sod Isoforms ◽  
No Bioavailability

Abstract Background Atherosclerotic plaques can elaborate reactive oxygen species (ROS) that reduce nitric oxide (NO) bioavailability. Cellular detoxification enzymes including various peroxiredoxin (PRDX) and superoxide dismutase (SOD) isoforms can inactivate ROS. The omega-3 fatty acid (n3-FA) eicosapentaenoic acid (EPA) reduced cardiovascular (CV) events in high-risk patients (REDUCE-IT), a benefit not observed with mixed n3-FAs containing docosahexaenoic acid (DHA). Purpose The purpose of this study was to compare the effects of EPA and DHA on NO bioavailability and expression of detoxification enzymes in the vascular endothelium in vitro. Methods Human umbilical vein endothelial cells (HUVECs) were pretreated with EPA or DHA at equimolar levels (10 μM) for 2 h, then challenged with IL-6 at 12 ng/ml for 24 h. Proteomic analysis was performed using LC/MS to measure relative protein expression. Only significant (p<0.05) changes between treatment groups >1-fold were analyzed. Cells were stimulated with calcium ionophore to measure NO and peroxynitrite (ONOO-) release using a porphyrinic nanosensor. Results EPA, but not DHA, augmented PRDX-2 and SOD1 expression in HUVECs relative to IL-6 alone (1.2-fold and 1.6-fold, respectively, p=0.03). EPA also significantly lowered other isoforms unlike DHA. Either EPA or DHA increased thioredoxin expression by 1.5-fold (p=0.001) and 1.3-fold (p=0.02), respectively and decreased SOD2 expression by 1.5-fold (p=8.75E-11) and 1.6-fold (p=6.03E-9), respectively. IL-6 alone only increased expression of 6 detoxification enzymes by at least 1.2-fold, relative to vehicle. Unlike DHA, EPA also increased the NO to ONOO- release ratio by 36% (p<0.05) relative to IL-6 alone, without changes in NO synthase (eNOS) expression. Conclusions n3-FAs differentially influenced NO bioavailability and expression of ROS detoxification proteins, including peroxiredoxin and SOD isoforms. The net benefits of EPA on eNOS function and ROS detoxification may contribute to reduced atherothrombotic risk compared to DHA. FUNDunding Acknowledgement Type of funding sources: Private company. Main funding source(s): Amarin Pharma Inc., Elucida Research

Abstract 1432: Nitric Oxide Bioavailability is Lower in Endothelium of Healthy Mexican American Donors as Compared to Non-Hispanic White Donors: Effect of Nebivolol

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
R. P Mason ◽  
Ruslan Kubant ◽  
Christopher Malinski ◽  
Adam Jacoby ◽  
Robert F Jacob ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Endothelial Function ◽  
Mexican Americans ◽  
Mexican American ◽  
Umbilical Vein ◽  
Coupling Efficiency ◽  
Calcium Ionophore ◽  
Stress Factors ◽  
No Bioavailability

Background: Epidemiologic studies indicate that Mexican Americans (MA) have a higher prevalence of CV risk factors and disease as compared with non-Hispanic whites (NHW). This increase in CV risk may be due, in part, to differences in endothelial function. In this study, we measured endothelial function in cells from normotensive, age-matched MA and NHW donors, as well as the effects of treatment with nebivolol, a new β 1 -selective blocker with vasodilating properties. Methods: Endothelial nitric oxide (NO) and peroxynitrite (ONOO − ) release in human umbilical vein endothelial cells (HUVECs) from age-matched MA and NHW donors were measured simultaneously using a nanosensor array. The effects of nebivolol on NO and ONOO − release were evaluated following pretreatment (24 h) with a calcium ionophore (CaI) as a receptor-independent stimulus. Endothelial NO synthase (eNOS) levels were measured by Western blot analysis, and drug-membrane interactions were determined by small angle x-ray diffraction approaches. Results: NO bioavailability in endothelial cells of MA donors was 30% lower than that of cells from NHW donors (383 ± 10 nM versus 543 ± 8 nM, n=6) following stimulation with CaI (1.0 μM). Pretreatment with nebivolol (1.0 μM) eliminated these interracial differences and enhanced NO release disproportionately in MA cells (57%) versus NHW cells (20%). Nebivolol also reduced ONOO − levels in MA endothelium by 75% (746 ± 12 nM to 195 ± 10 nM) and by 50% in NHW cells (416 ± 7 nM to 191 ± 13 nM). The ratio of NO to ONOO − , an indicator of eNOS coupling, increased more than 5-fold in MA cells following nebivolol treatment. In addition, eNOS levels were 40% lower in MA endothelium compared to NHW, but increased 2-fold with nebivolol treatment. These effects were not observed with atenolol, a hydrophilic β 1 -selective antagonist. Conclusion: We observed differences between Mexican Americans and non-Hispanic whites in endothelial NO bioavailability and nitroxidative stress–factors that may contribute to increased CV risk. Treatment with nebivolol, but not atenolol, enhanced both the expression and coupling efficiency of eNOS in Mexican American endothelium.

scholarly journals Alpha-Asarone Protects Endothelial Cells from Injury by Angiotensin II

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Hai-Xia Shi ◽  
Jiajun Yang ◽  
Tao Yang ◽  
Yong-Liang Xue ◽  
Jun Liu ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Angiotensin Ii ◽  
Cell Injury ◽  
Fluorescent Dyes ◽  
Umbilical Vein ◽  
Protective Effects ◽  
Ang Ii ◽  
Versus Model

α-Asarone is the major therapeutical constituent ofAcorus tatarinowiiSchott. In this study, the potential protective effects ofα-asarone against endothelial cell injury induced by angiotensin II were investigatedin vitro. The EA.hy926 cell line derived from human umbilical vein endothelial cells was pretreated withα-asarone (10, 50, 100 µmol/L) for 1 h, followed by coincubation with Ang II (0.1 µmol/L) for 24 h. Intracellular nitric oxide (NO) and reactive oxygen species (ROS) were detected by fluorescent dyes, and phosphorylation of endothelial nitric oxide synthase (eNOS) atSer1177was determined by Western blotting.α-Asarone dose-dependently mitigated the Ang II-induced intracellular NO reduction (P<0.01versus model) and ROS production (P<0.01versus model). Furthermore, eNOS phosphorylation (Ser1177) by acetylcholine was significantly inhibited by Ang II, while pretreatment for 1 h withα-asarone partially prevented this effect (P<0.05versus model). Additionally, cell viability determined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay (105~114.5% versus control,P>0.05) was not affected after 24 h of incubation withα-asarone at 1–100 µmol/L. Therefore,α-asarone protects against Ang II-mediated damage of endothelial cells and may be developed to prevent injury to cardiovascular tissues.

Effect of hypoxia on adherence of granulocytes to endothelial cells in vitro

1994 ◽  
Vol 267 (3) ◽  
pp. H874-H879 ◽  
Author(s):  
A. Pietersma ◽  
N. De Jong ◽  
J. F. Koster ◽  
W. Sluiter
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Calcium Ionophore ◽  
Intercellular Adhesion Molecule ◽  
Protective Mechanism ◽  
Hypoxic Conditions ◽  
Umbilical Vein Endothelial Cells ◽  
Stimulation Of

The objective of this study was to investigate the effect of hypoxia on the adhesiveness of endothelial cells for granulocytes. Human umbilical vein endothelial cells (HUVEC) were exposed to a PO2 of 7.5 mmHg (1.0 kPa), and the adherence of granulocytes was assessed under continuous hypoxia by means of a hypoxic incubator room. After 2 h of hypoxia the adherence of granulocytes decreased to 50% of the normoxic control, which was not due to a decreased viability of the endothelial cells nor to an increased generation of the antiadhesive factors nitric oxide, prostacyclin, and adenosine. Hypoxia also had no effect on the expression of intercellular adhesion molecule (ICAM)-1 or ICAM-2 on the endothelium. Although the mechanism of the action of hypoxia on the adhesiveness of endothelial cells remains unclear as yet, our data suggest that HUVEC possess a protective mechanism that prevents granulocyte adherence to endothelial cells under extreme hypoxic conditions. The decreased adherence seems paradoxical to the in vivo situation for which the increased margination of granulocytes within the vascular compartment of the ischemic tissue has been observed. However, hypoxia did not impair the potential adhesiveness of HUVEC, since stimulation of endothelial cells under hypoxic conditions with calcium ionophore or lipopolysaccharide increased the adherence of granulocytes in a similar fashion as under normoxic conditions. We therefore conclude that the increased margination of granulocytes during ischemia may be accomplished by the additional stimulation of hypoxic endothelial cells.

scholarly journals Mitigation of Particulate Matter-Induced Inflammation and Vasoactivity in Human Vascular Endothelial Cells by Omega-3 Polyunsaturated Fatty Acids

2018 ◽  
Vol 15 (10) ◽  
pp. 2293
Author(s):  
Jaya Sriram ◽  
Olorunfemi Adetona ◽  
Tonya Orchard ◽  
Chieh-Ming Wu ◽  
James Odei
Keyword(s):  
Fatty Acids ◽  
Endothelial Cells ◽  
Particulate Matter ◽  
Polyunsaturated Fatty Acids ◽  
Fish Oil ◽  
Umbilical Vein ◽  
Diesel Exhaust Particles ◽  
Omega 3 ◽  
Oil Formulation

Airborne particulate matter (PM) exposure remains the leading environmental risk factor for disease globally. Interventions to mitigate the adverse effects of PM are required, since there is no discernible threshold for its effects, and exposure reduction approaches are limited. The mitigation of PM (specifically diesel exhaust particles (DEP))-induced release of pro-inflammatory cytokines interleukin-6 (IL-6) and interleukin-8 (IL-8) and vasoconstrictor endothelin-1 (ET-1) after 24 and 48 h of exposure by pre-treatment with individual pure, combined pure, and an oil formulation of two fish oil omega-3 polyunsaturated fatty acids (ω-3 PUFAs), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA) were all tested at an equivalent concentration of 100 µM in vitro in human umbilical vein endothelial cells. The PUFAs and fish oil formulation completely mitigated or diminished the DEP-induced release of IL-6, IL-8, and ET-1 by 14–78%. DHA was more effective in reducing the levels of the DEP-induced release of the cytokines, especially IL-6 after 48 h of DEP exposure in comparison to EPA (p < 0.05), whereas EPA seemed to be more potent in reducing ET-1 levels. The potential of fish ω-3 PUFAs to mitigate PM-induced inflammation and vasoactivity was demonstrated by this study.

Monocyte-derived microparticles and exosomes induce procoagulant and apoptotic effects on endothelial cells

2008 ◽  
Vol 100 (05) ◽  
pp. 878-885 ◽  
Author(s):  
Tal Tamari ◽  
Benjamin Brenner ◽  
Anat Aharon
Keyword(s):  
Endothelial Cells ◽  
Tissue Factor ◽  
Umbilical Vein ◽  
Calcium Ionophore ◽  
Tube Formation ◽  
Ionophore A23187 ◽  
Membrane Phospholipids ◽  
Flip Flop ◽  
Time Activity

SummaryMicrovesicles (MVs) which include microparticles (MPs) and exosomes are found in blood circulation in normal physiologic conditions and are increased in a variety of diseases. This study evaluated the effects of MVs on human umbilical vein endothelial cells (HUVEC) by morphologic changes, apoptosis, and thrombogenicty, in vitro. Stimulation of monocyte cell line (THP-1) by starvation or by endotoxin and calcium ionophore A23187 resulted in the release of MVs which express exosome marker Tsg 101, negative phospholipids in their leaflets, monocyte markers (CD18, CD14) and active tissue factor (TF). MVs were found to disrupt EC integrity and rapidly induce membrane blebbing. Brief exposure (2–4 hours) to MVs resulted in EC membrane phospholipids “flip-flop” while longer stimulation (20 hours) led to two contradicting outcomes – tube formation as well as apoptosis, as assessed by nuclear fragmentation. Additionally, MVs exposure resulted in increased cell surface thrombogenicity and perturbation of the endothelial haemostatic balance, which were enhanced during longer exposure time. Activity, antigen level and mRNA expression of the coagulation initiator TF were elevated due to (i) adherence of MVs derived TF to the EC membrane, and (ii) an increase in endothelial TF expression. Furthermore, levels of the anticoagulant tissue factor pathway inhibitor (TFPI) and thrombomodulin (TM) were decreased. These findings demonstrate that monocyte MVs increase endothelial thrombogenicity and apoptosis. In addition, they induce tube formation which may indicate their angiogenic effect. These findings may clarify, in part, the role of MVs in EC dysfunction associated with inflammatory diseases and hypercoagulable states.

Nitric oxide synthesis is impaired in glutathione-depleted human umbilical vein endothelial cells

1993 ◽  
Vol 265 (3) ◽  
pp. C728-C732 ◽  
Author(s):  
D. Ghigo ◽  
P. Alessio ◽  
A. Foco ◽  
F. Bussolino ◽  
C. Costamagna ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Reduced Glutathione ◽  
Umbilical Vein ◽  
Calcium Ionophore ◽  
Human Endothelial Cells ◽  
Cgmp Production ◽  
Endothelium Derived Relaxing Factor ◽  
Citrulline Synthesis ◽  
Umbilical Vein Endothelial Cells

Human endothelial cells cultured from umbilical vein (HUVEC) were tested for their ability to synthesize nitric oxide (NO), which has been identified as an endothelium-derived relaxing factor. The synthesis of this free radical (detected as citrulline, which is produced stoichiometrically with NO from arginine) in HUVEC is Ca2+ dependent, is increased sevenfold by the calcium ionophore ionomycin, and accounts for most basal and ionomycin-induced guanosine 3',5'-cyclic monophosphate (cGMP) production. Loading of cells with reduced glutathione (GSH), but not with N-(2-mercaptopropionyl)- glycine (MPG), led to increased citrulline production, both basally and after ionomycin stimulation. When the cells were depleted of GSH by incubation with 1-chloro-2,4-dinitrobenzene (CDNB), citrulline synthesis and cGMP production were inhibited in a concentration-dependent way. CDNB was not cytotoxic and did not inhibit cGMP increase elicited by sodium nitroprusside; cell loading with GSH (but not with MPG) relieved the block of citrulline synthesis. These results suggest that GSH is necessary in HUVEC for NO synthesis rather than for the NO effect on guanylate cyclase.

scholarly journals VLDL Induced Modulation of Nitric Oxide Signalling and Cell Redox Homeostasis in HUVEC

2017 ◽  
Vol 2017 ◽  
pp. 1-15 ◽  
Author(s):  
Maria Chiara Magnifico ◽  
Roxana Elena Oberkersch ◽  
Azzurra Mollo ◽  
Luca Giambelli ◽  
Yasmine Grooten ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Umbilical Vein ◽  
Redox Homeostasis ◽  
No Synthase ◽  
Very Low Density Lipoproteins ◽  
Enos Uncoupling ◽  
Phosphorylation Pattern ◽  
No Bioavailability ◽  
Umbilical Vein Endothelial Cells

High levels of circulating lipoprotein constitute a risk factor for cardiovascular diseases, and in this context, the specific role of the very-low-density lipoproteins (VLDL) is poorly understood. The response of human umbilical vein endothelial cells (HUVEC) to VLDL exposure was studied, especially focusing on the pathways involved in alteration of redox homeostasis and nitric oxide (NO) bioavailability. The results obtained by the analysis of the expression level of genes implicated in the NO metabolism and oxidative stress response indicated a strong activation of inducible NO synthase (iNOS) upon 24 h exposure to VLDL, particularly if these have been preventively oxidised. Simultaneously, both mRNA and protein expression of endothelial NO synthase (eNOS) were decreased and its phosphorylation pattern, at the key residues Tyr495 and Ser1177, strongly suggested the occurrence of the eNOS uncoupling. The results are consistent with the observed increased production of nitrites and nitrates (NOx), reactive oxygen species (ROS), 3-nitrotyrosine (3-NT), and, at mitochondrial level, a deficit in mitochondrial O2consumption. Altogether, these data suggest that the VLDL, particularly if oxidised, when allowed to persist in contact with endothelial cells, strongly alter NO bioavailability, affecting redox homeostasis and mitochondrial function.

Regional differentiation in the rat aorta for a novel signaling pathway: leucine to glutamate

1997 ◽  
Vol 273 (3) ◽  
pp. H1484-H1492 ◽  
Author(s):  
D. Schachter ◽  
J. C. Sang
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Human Peripheral Blood ◽  
Umbilical Vein ◽  
Rat Aorta ◽  
Signal Pathway ◽  
Regional Differentiation ◽  
Differentiation Pattern ◽  
Cells Cultured

Rat aortic endothelium is differentiated regionally for regulating guanosine 3',5'-cyclic monophosphate (cGMP) levels in underlying smooth muscle by signaling via nitric oxide and prostaglandin H2. Highest activity is just distal to the aortic arch and diminishes peripherally. The same differentiation pattern is reported here for a third and novel signal pathway: endothelial conversion of L-leucine to L-glutamate. Sequential segments of rat aorta incubated in vitro convert L-[U-14C]leucine to a major 14C metabolite identified as L-glutamate. Net synthesis of glutamate is greatest in aortic segments of the "windkessel" region; significant quantities are also observed in the pancreas, testis, and lung but very little in 10 additional tissues. Endothelial cells cultured from mouse brain, human umbilical vein, or bovine aorta and human peripheral blood macrophages also form [14C]glutamate. When aortic segments are denuded of endothelium, treatment with L-glutamate in the presence of 3-isobutyl-1-methylxanthine significantly increases the cGMP content. A number of leucine derivatives inhibit the leucine-to-glutamate conversion and decrease the cGMP content in aortic segments in vitro.

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