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.

scholarly journals Pretreatment with Astragaloside IV protects human umbilical vein endothelial cells from hydrogen peroxide induced oxidative stress and cell dysfunction via inhibiting eNOS uncoupling and NADPH oxidase – ROS – NF-κB pathway

2016 ◽  
Vol 94 (11) ◽  
pp. 1132-1140 ◽  
Author(s):  
Chonghua Xu ◽  
Futian Tang ◽  
Meili Lu ◽  
Jing Yang ◽  
Ronghui Han ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Nitric Oxide ◽  
Hydrogen Peroxide ◽  
Endothelial Cells ◽  
Nadph Oxidase ◽  
Protein Expression ◽  
Umbilical Vein ◽  
Astragaloside Iv ◽  
Enos Uncoupling ◽  
Umbilical Vein Endothelial Cells

Endothelial cell injury caused by reactive oxygen species (ROS) plays a critical role in the pathogenesis of cardiovascular disorders. Astragaloside IV (AsIV) possesses potent antioxidant properties against oxidative stress through undefined mechanism(s). We sought to investigate whether AsIV protects human umbilical vein endothelial cells (HUVECs) from hydrogen peroxide (H2O2) induced oxidative stress focusing on eNOS uncoupling and the NADPH oxidase – ROS – NF-κB pathway. Compared with HUVECs incubated with H2O2 alone, pretreatment with AsIV significantly increased the viability of HUVECs, which was accompanied with apparent increase in nitric oxide (NO) production and decrease in intracellular superoxide anion production. Furthermore, pretreatment with AsIV increased endothelial nitric oxide synthase (eNOS) dimer/monomer ratio and its critical cofactor tetrahydrobiopterin (BH4) content, decreased Nox4 protein expression (the most abundant Nox isoform in HUVECs), inhibited translocation of NF-κB p65 subunit into nuclear fraction while enhanced the protein expression of IκB-α (the inhibitor of NF-κB p65), reduced the levels of IL-1β, IL-6, and TNF-α in HUVECs medium, and decreased iNOS protein expression. These results suggest that AsIV may protect HUVECs from H2O2-induced oxidative stress via inhibiting NADPH oxidase – ROS – NF-κB pathway and eNOS uncoupling.

Hypoxia-induced pulmonary endothelin-1 expression is unaltered by nitric oxide

2002 ◽  
Vol 92 (3) ◽  
pp. 1152-1158 ◽  
Author(s):  
Scott Earley ◽  
Leif D. Nelin ◽  
Louis G. Chicoine ◽  
Benjimen R. Walker
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Promoter Activity ◽  
Umbilical Vein ◽  
Hypoxia Inducible Factor ◽  
Hypoxic Exposure ◽  
Mrna Levels ◽  
Protective Effects ◽  
No Donor ◽  
Umbilical Vein Endothelial Cells

Nitric oxide (NO) attenuates hypoxia-induced endothelin (ET)-1 expression in cultured umbilical vein endothelial cells. We hypothesized that NO similarly attenuates hypoxia-induced increases in ET-1 expression in the lungs of intact animals and reasoned that potentially reduced ET-1 levels may contribute to the protective effects of NO against the development of pulmonary hypertension during chronic hypoxia. As expected, hypoxic exposure (24 h, 10% O2) increased rat lung ET-1 peptide and prepro-ET-1 mRNA levels. Contrary to our hypothesis, inhaled NO (iNO) did not attenuate hypoxia-induced increases in pulmonary ET-1 peptide or prepro-ET-1 mRNA levels. Because of this surprising finding, we also examined the effects of NO on hypoxia-induced increases in ET peptide levels in cultured cell experiments. Consistent with the results of iNO experiments, administration of the NO donor S-nitroso- N-acetyl-penicillamine to cultured bovine pulmonary endothelial cells did not attenuate increases in ET peptide levels resulting from hypoxic (24 h, 3% O2) exposure. In additional experiments, we examined the effects of NO on the activity of a cloned ET-1 promoter fragment containing a functional hypoxia inducible factor-1 binding site in reporter gene experiments. Whereas moderate hypoxia (24 h, 3% O2) had no effect on ET-1 promoter activity, activity was increased by severe hypoxic (24 h, 0.5% O2) exposure. ET-1 promoter activity after S-nitroso- N-acetyl-penicillamine administration during severe hypoxia was greater than that in normoxic controls, although activity was reduced compared with that in hypoxic controls. These findings suggest that hypoxia-induced pulmonary ET-1 expression is unaffected by NO.

scholarly journals Cilostazol Induces eNOS and TM Expression via Activation with Sirtuin 1/Krüppel-like Factor 2 Pathway in Endothelial Cells

2021 ◽  
Vol 22 (19) ◽  
pp. 10287
Author(s):  
Chih-Hsien Wu ◽  
Yi-Lin Chiu ◽  
Chung-Yueh Hsieh ◽  
Guo-Shiang Tsung ◽  
Lian-Shan Wu ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Endothelial Nitric Oxide Synthase ◽  
Umbilical Vein ◽  
Sirtuin 1 ◽  
No Production ◽  
Beneficial Effects ◽  
Umbilical Vein Endothelial Cells ◽  
Oxide Synthase ◽  
Endothelial Nitric Oxide

Cilostazol was suggested to be beneficial to retard in-stent atherosclerosis and prevent stent thrombosis. However, the mechanisms responsible for the beneficial effects of cilostazol are not fully understood. In this study, we attempted to verify the mechanism of the antithrombotic effect of cilostazol. Human umbilical vein endothelial cells (HUVECs) were cultured with various concentrations of cilostazol to verify its impact on endothelial cells. KLF2, silent information regulator transcript-1 (SIRT1), endothelial nitric oxide synthase (eNOS), and endothelial thrombomodulin (TM) expression levels were examined. We found cilostazol significantly activated KLF2 expression and KLF2-related endothelial function, including eNOS activation, Nitric oxide (NO) production, and TM secretion. The activation was regulated by SIRT1, which was also stimulated by cilostazol. These findings suggest that cilostazol may be capable of an antithrombotic and vasculoprotective effect in endothelial cells.

scholarly journals Effects of Propofol on Cyclic Strain-induced Endothelin-1 Expression in Human Umbilical Vein Endothelial Cells

2009 ◽  
Vol 110 (1) ◽  
pp. 74-80 ◽  
Author(s):  
Tzu-Hurng Cheng ◽  
Jin-Jer Chen ◽  
Cheng-Hsien Chen ◽  
Kar-Lok Wong
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Protein Kinase ◽  
Umbilical Vein ◽  
Cyclic Strain ◽  
Nitric Oxide Production ◽  
Human Umbilical Vein ◽  
Oxide Production ◽  
Extracellular Signal ◽  
Umbilical Vein Endothelial Cells

Background Propofol is one of the most popular intravenous induction agents of general anesthesia. Experimental results revealed that propofol exerted hypotensive and antioxidative effects. However, the intracellular mechanism of propofol remains to be delineated. The aims of this study were to test the hypothesis that propofol may alter strain-induced endothelin-1 (ET-1) secretion and nitric oxide production, and to identify the putative underlying signaling pathways in human umbilical vein endothelial cells. Methods Cultured human umbilical vein endothelial cells were exposed to cyclic strain in the presence of propofol, and ET-1 expression was examined by Northern blotting and enzyme-linked immunosorbent assay kit. Activation of extracellular signal-regulated protein kinase, endothelial nitric oxide synthase, and protein kinase B were assessed by Western blot analysis. Results The authors show that propofol inhibits strain-induced ET-1 expression, strain-increased reactive oxygen species formation, and extracellular signal-regulated protein kinase phosphorylation. On the contrary, nitric oxide production, endothelial nitric oxide synthase activity, and protein kinase B phosphorylation were enhanced by propofol treatment. Furthermore, in the presence of PTIO, a nitric oxide scavenger, and KT5823, a specific inhibitor of cyclic guanosine monophosphate-dependent protein kinase, the inhibitory effect of propofol on strain-induced extracellular signal-regulated protein kinase phosphorylation and ET-1 release was reversed. Conclusions The authors demonstrate for the first time that propofol inhibits strain-induced ET-1 secretion and enhances strain-increased nitric oxide production in human umbilical vein endothelial cells. Thus, this study delivers important new insight into the molecular pathways that may contribute to the proposed hypotensive effects of propofol in the cardiovascular system.

Paradoxical overexpression and translocation of connexin43 in homocysteine-treated endothelial cells

2002 ◽  
Vol 282 (6) ◽  
pp. H2124-H2133 ◽  
Author(s):  
Hong Li ◽  
Sergey Brodsky ◽  
Sindu Kumari ◽  
Virginijus Valiunas ◽  
Peter Brink ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Fluorescent Protein ◽  
Lucifer Yellow ◽  
Umbilical Vein ◽  
Expression Screening ◽  
Fluorescent Markers ◽  
Junctional Communication ◽  
Green Fluorescent ◽  
Umbilical Vein Endothelial Cells

Hyperhomocysteinemia is an established cause of defective vasorelaxation. Gene expression screening of human umbilical vein endothelial cells (HUVEC) treated with homocysteine (Hcy) revealed that connexin43 (Cx43) was upregulated. Expression of Cx43 was increased more than twofold in Hcy-treated HUVEC. Gap junctional communication (Lucifer yellow and whole cell patch clamp) was not enhanced in Hcy-treated HUVEC. HUVEC expressing chimeric Cx43-green fluorescent protein exhibited it at cell-cell contacts in control but showed redistribution to the intracellular compartment(s) in Hcy-treated cells. Confocal microscopy of HUVEC stained with anti-Cx43, mitochondrial, and endoplasmic reticulum fluorescent markers showed the localization of Cx43 to the plasma membrane of control cells and its colocalization with the mitochondrial marker in Hcy-treated HUVEC. Studies of isolated mitochondria confirmed overexpression of Cx43 in the mitochondria of Hcy-treated HUVEC. Microdissected renal interlobar arteries, which normally exhibit endothelium-derived hyperpolarizing factor-induced vasorelaxation, showed reduced nitric oxide synthase- and cyclooxygenase-independent vasorelaxation to acetylcholine after pretreatment with Hcy. In summary, Hcy-induced upregulation of Cx43 transcript and protein expression are associated with unaltered intercellular communication, redistribution of Cx43 in HUVEC, and reduced nitric oxide- and prostanoid-independent vascular responses to acetylcholine in Hcy-treated arteries.

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.

Sign in / Sign up

Export Citation Format

Share Document