scholarly journals Hydrogen-Rich Medium Ameliorates Lipopolysaccharides-Induced Mitochondrial Fission and Dysfunction in Human Umbilical Vein Endothelial Cells (Huvecs) via Up-Regulating HO-1 Expression

2021 ◽  
Author(s):  
Keliang Xie ◽  
Xing Mao ◽  
Naqi Lian ◽  
Yanyan Wang ◽  
Yuzun Wang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Umbilical Vein ◽  
Mitochondrial Fission ◽  
Protective Effects ◽  
Atp Content ◽  
Rich Medium ◽  
Umbilical Vein Endothelial Cells

Abstract Background Sepsis is defined as life-threatening organ dysfunction caused by a dysregulated host response to infection. It has been showed that the change of mitochondrial dynamics has been proved to be one of the main causes of death in patients with severe sepsis. And hydrogen has been proved to exert its protective effects against sepsis via heme oxygenase-1 (HO-1). This study was designed to demonstrate that whether the benefit effects of hydrogen can maintain the dynamic process of mitochondrial fusion/fission to mitigate human umbilical vein endothelial cells (HUVECs) injury exposed to endotoxin through HO-1. Methods HUVECs cells cultured with medium which contained Lipopolysaccharides (LPS), Saline, hydrogen, Mdivi-1 (a dynamin-related protein 1 [Drp1] inhibitor) or zinc protoporphyrin Ⅸ (Znpp) (a HO-1 inhibitor) were also used in the research. Cell death and apoptosis were assessed using FITC annexin V and PI. Mitochondria were stained with Mitotracker orange and observed by confocal microscope. Oxygen consumption rate was assessed by seahorse xf24 extracellular analyzer. Mitochondrial membrane potential monitored by JC-1 dye. The expressions of Drp1 and HO-1 were tested by Western blot. The co-localization of Drp1 and mitochondria was determined by immunofluorescence. Results LPS caused a decrease in ATP content, mitochondrial membrane potential, and maximal respiration rate. At the same time, increased expression of Drp1 were observed in LPS-stimulated HUVECs, concomitantly with excessive mitochondrial fission. We found that hydrogen-rich medium can increase ATP content, mitochondrial membrane potential and maximal respiration rate, and decrease the expression of Drp1 in LPS-treated HUVECs. Meanwhile, hydrogen can ameliorate excessive mitochondrial fission caused by LPS. Furthermore, hydrogen-rich medium had a similar effect to Mdivi-1, a mitochondrial fission blocker. Both of them rescued the up-regulation of Drp1 and mitochondrial fission induced by LPS, then normalized mitochondrial shape after LPS stimulation. But after Znpp pretreatment, HO-1 expression was inhibited and the protective effects of hydrogen were abrogated. Conclusions Hydrogen-rich medium can alleviate the LPS-induced mitochondrial fusion/fission and dysfunction in HUVECs via HO-1 up-regulation.

Protective effects and plausible mechanisms of antler-velvet polypeptide against hydrogen peroxide induced injury in human umbilical vein endothelial cells

2017 ◽  
Vol 95 (5) ◽  
pp. 610-619 ◽  
Author(s):  
Wenhe Zhu ◽  
Huiyan Wang ◽  
Wei Zhang ◽  
Na Xu ◽  
Junjie Xu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Flow Cytometry ◽  
Endothelial Cells ◽  
Membrane Potential ◽  
Cell Viability ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Umbilical Vein ◽  
Protective Effects ◽  
Umbilical Vein Endothelial Cells

Antler velvet polypeptide (VAP) is a prominent bioactive component of antler velvet. Whereas uncharacterized crude extracts have typically been used in pharmacological studies, in this study, the velvet polypeptide was isolated and purified by acid water extraction, ethanol precipitation, ammonium sulfate fractionation and precipitation, and chromatography, progressively. Human umbilical vein endothelial cells (HUVECs) were induced with H2O2 followed purified polypeptide treatment. Cell viability was evaluated by MTT assay. The apoptosis of cells was detected by fluorescence microscopy and flow cytometry. A cell analyzer was used to measure the mitochondrial membrane potential. The intracellular reactive oxygen species (ROS) levels were determined by flow cytometry. Oxidative stress related biochemical parameters were detected, and the expression of apoptosis-related proteins was examined by Western blot analysis. The results indicated that a 7.0 kDa polypeptide (VAP II) was isolated from antler velvet. VAP II enhanced cell viability, decreased cell apoptosis, reversed depolarization of mitochondrial membrane potential, decreased ROS levels, inhibited oxidative stress, and regulated the downstream signaling apoptotic cascade expression caused by H2O2. The protective effects of VAP II on HUVECs suggests a potential strategy for the treatment of cardiovascular disease.

Flow- and bradykinin-induced nitric oxide production by endothelial cells is independent of membrane potential

1996 ◽  
Vol 270 (2) ◽  
pp. C546-C551 ◽  
Author(s):  
K. J. Gooch ◽  
J. A. Frangos
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Membrane Potential ◽  
Umbilical Vein ◽  
Fold Increase ◽  
No Production ◽  
Control Medium ◽  
Rich Medium ◽  
Membrane Hyperpolarization ◽  
Umbilical Vein Endothelial Cells

The objective of this study was to evaluate the role transmembrane potential plays in flow-induced nitric oxide (NO) production in endothelial cells (EC). NO production was monitored by measuring intracellular guanosine 3',5'-cyclic monophosphate (cGMP) and extracellular nitrite plus nitrate (NOx). Primary human umbilical vein endothelial cells (HUVEC) were exposed to laminar flow (22 dyn/cm2) of medium with 5.4 mM KCl (control medium) with or without 3 mM tetraethylammonium chloride (TEA) or 90 mM KCl (K(+)-rich medium). Bradykinin (BK) was added to time-matched stationary cultures to give a final concentration of 5 nM. With control medium, 30 s, 2 min, and 3 h of treatment with flow or 2 min of treatment with BK resulted in an approximately threefold increase in cGMP over stationary cultures. Depolarization with KCl or TEA did not influence cGMP production in flow-treated or stationary cultures. Flow of either control or potassium-rich medium resulted in an approximately 10-fold increase in average NOx production rate over 3 h compared with stationary cultures. Taken together these data indicate that neither membrane hyperpolarization nor normal membrane potential is necessary for flow- or BK-induced NO production by HUVEC.

scholarly journals Zhengyuan prescription inhibits X-ray-induced injury of human umbilical vein endothelial cells by activating the Nrf2 signaling pathway

2021 ◽  
Author(s):  
cheng xiaoni ◽  
Yalei Pan ◽  
Zhishu Tang ◽  
Rui Zhou ◽  
Haichao Zhang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Endothelial Cells ◽  
Membrane Potential ◽  
Mitochondrial Membrane ◽  
Umbilical Vein ◽  
X Rays ◽  
X Ray ◽  
Nrf2 Signaling Pathway ◽  
Umbilical Vein Endothelial Cells ◽  
And Oxidative Stress

Abstract Background: Zhengyuan prescription (ZYP) is a Chinese herbal medicine used in clinical practice to protect against radiotherapy-induced injuries. In this study, we investigate the protective effect of ZYP against X-ray-induced injury of human umbilical vein endothelial cells (HUVECs), and we explore the mechanisms underlying this effect.Methods: After 3 h of ZYP intervention, the cells in the ZYP group were irradiated with 6 Gy X-rays and cultured for 48 h. Subsequently, the cell viability, cell morphology, mitochondrial membrane potential, and apoptosis and oxidative stress markers were observed, as well as the expressions of apoptotic and oxidative stress proteins.Results: The obtained results demonstrate that exposure to X-rays promotes cell death, reduces mitochondrial membrane potential, and induces the pirroduction of intracellular reactive oxygen species (ROS). Pretreatment with ZYP reverses these effects to a great extent. Moreover, it up-regulates the expression of the B-cell lymphoma 2 (Bcl-2) apoptosis inhibitor protein while down-regulating the expressions of Bcl-2-associated X protein (Bax), caspase-3, and caspase-9. Interestingly, ZYP can also inhibit oxidative stress injury by activating the expression of Nrf2 (Nuclear Factor E2 related factor) regulated antioxidant enzyme genes such as Heme oxygenase 1 (HO-1) and NAD(P)H:quinone oxidoreductase-1 (NQO1)Conclusions: This study is the first to demonstrate that ZYP suppresses X-ray-induced injury of HUVECs by activating the Nrf2 signaling pathway.

scholarly journals Beta vulgaris rubra L. (Beetroot) Peel Methanol Extract Reduces Oxidative Stress and Stimulates Cell Proliferation via Increasing VEGF Expression in H2O2 Induced Oxidative Stressed Human Umbilical Vein Endothelial Cells

Genes ◽  
2021 ◽  
Vol 12 (9) ◽  
pp. 1380
Author(s):  
Laila Naif Al-Harbi ◽  
Subash-Babu Pandurangan ◽  
Alhanouf Mohammed Al-Dossari ◽  
Ghalia Shamlan ◽  
Ahmad Mohammad Salamatullah ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Proliferation ◽  
Endothelial Cells ◽  
Membrane Potential ◽  
Cell Growth ◽  
Mitochondrial Membrane ◽  
Methanol Extract ◽  
Umbilical Vein ◽  
Vascular Cell ◽  
Umbilical Vein Endothelial Cells

The antioxidant capacity of polyphenols and flavonoids present in dietary agents aids in arresting the development of reactive oxygen species (ROS) and protecting endothelial smooth muscle cells from oxidative stress/induced necrosis. Beetroot (Beta vulgaris var. rubra L.; BVr) is a commonly consumed vegetable representing a rich source of antioxidants. Beetroot peel’s bioactive compounds and their role in human umbilical vein endothelial cells (HUVECs) are still under-researched. In the present study, beetroot peel methanol extract (BPME) was prepared, and its effect on the bio-efficacy, nuclear integrity, mitochondrial membrane potential and vascular cell growth, and immunoregulation-related gene expression levels in HUVECs with induced oxidative stress were analysed. Gas chromatography–mass spectroscopy (GC-MS) results confirmed that BPME contains 5-hydroxymethylfurfural (32.6%), methyl pyruvate (15.13%), furfural (9.98%), and 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-Pyran-4-one (12.4%). BPME extract effectively enhanced cell proliferation and was confirmed by MTT assay; the nuclear integrity was confirmed by propidium iodide (PI) staining assay; the mitochondrial membrane potential (Δψm) was confirmed by JC-1 staining assay. Annexin V assay confirmed that BPME-treated HUVECs showed 99% viable cells, but only 39.8% viability was shown in HUVECs treated with H2O2 alone. In addition, BPME treatment of HUVECs for 48 h reduced mRNA expression of lipid peroxide (LPO) and increased NOS-3, Nrf-2, GSK-3β, GPX, endothelial nitric oxide synthase (eNOS) and vascular cell growth factor (VEGF) mRNA expression levels. We found that BPME treatment decreased proinflammatory (nuclear factor-κβ (F-κβ), tissue necrosis factor-α (TNF-α), toll-like receptor-4 (TLR-4), interleukin-1β (IL-1β)) and vascular inflammation (intracellular adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), EDN1, IL-1β)-related mRNA expressions. In conclusion, beetroot peel treatment effectively increased vascular smooth cell growth factors and microtubule development, whereas it decreased vascular inflammatory regulators. BPME may be beneficial for vascular smooth cell regeneration, tissue repair and anti-ageing potential.

scholarly journals Ethanol inhibits monocyte chemotactic protein-1 expression in interleukin-1β-activated human endothelial cells

2005 ◽  
Vol 289 (4) ◽  
pp. H1669-H1675 ◽  
Author(s):  
John P. Cullen ◽  
Shariq Sayeed ◽  
Ying Jin ◽  
Nicholas G. Theodorakis ◽  
James V. Sitzmann ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Binding Activity ◽  
Protective Effects ◽  
Monocyte Adhesion ◽  
Monocyte Chemotactic Protein ◽  
Chemotactic Protein ◽  
Subendothelial Space ◽  
Umbilical Vein Endothelial Cells

The aim of this study was to determine the effect of ethanol (EtOH) on endothelial monocyte chemotactic protein-1 (MCP-1) expression. IL-1β increased the production of MCP-1 by human umbilical vein endothelial cells from undetectable levels to ∼900 pg/ml at 24 h. EtOH dose-dependently inhibited IL-1β-stimulated MCP-1 secretion as determined by ELISA: 25 ± 1%, 35 ± 7%, and 65 ± 5% inhibition for 1, 10, and 100 mM EtOH, respectively, concomitant with inhibition of monocyte adhesion to activated endothelial cells. Similarly, EtOH dose-dependently inhibited IL-1β-stimulated MCP-1 mRNA expression. Experiments with actinomycin D demonstrated that EtOH decreased the stability of MCP-1 mRNA. In addition, EtOH significantly reduced NF-κB and AP-1 binding activity induced by IL-1β and inhibited MCP-1 gene transcription. Binding of 125I-labeled MCP-1 to its receptor (CCR2) on THP-1 human monocytic cells was not affected by EtOH treatment. Modulation of the expression of MCP-1 represents a mechanism whereby EtOH could inhibit atherogenesis by blocking the crucial early step of monocyte adhesion and subsequent recruitment to the subendothelial space. These actions of EtOH may underlie, in part, its cardiovascular protective effects in vivo.

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 Glyoxal Damages Human Aortic Endothelial Cells by Perturbing the Glutathione, Mitochondrial Membrane Potential, and Mitogen-Activated Protein Kinase Pathways

2021 ◽  
Author(s):  
Ming-Zhang Xie ◽  
Chun Guo ◽  
Jia-Qi Dong. BA ◽  
Jie Zhang ◽  
Ke-Tao Sun ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Membrane Potential ◽  
Protein Kinase ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Mitogen Activated Protein Kinase ◽  
Aortic Endothelial Cells ◽  
Mitogen Activated Protein ◽  
Human Aortic Endothelial Cells ◽  
Quantitative Fluorescence

Abstract Background: Exposure to glyoxal, the smallest dialdehyde, is associated with several diseases; humans are routinely exposed to glyoxal because of its ubiquitous presence in foods and the environment. The aim of this study was to examine the damage caused by glyoxal in human aortic endothelial cells. Methods: Cell survival assays and quantitative fluorescence assays were performed to measure DNA damage; oxidative stress was detected by colorimetric assays and quantitative fluorescence, and the mitogen-activated protein kinase pathways were assessed using western blotting. Results: Exposure to glyoxal was found to be linked to abnormal glutathione activity, the collapse of mitochondrial membrane potential, and the activation of mitogen-activated protein kinase pathways. However, DNA damage and thioredoxin oxidation were not induced by dialdehydes. Conclusions: Intracellular glutathione, members of the mitogen-activated protein kinase pathways, and the mitochondrial membrane potential are all critical targets of glyoxal. These findings provide novel insights into the molecular mechanisms perturbed by glyoxal and may facilitate the development of new therapeutics and diagnostic markers for cardiovascular diseases.

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