scholarly journals Characterization of the interactions between procoagulant albumin and human endothelial cells

Blood ◽  
1993 ◽  
Vol 82 (9) ◽  
pp. 2684-2692
Author(s):  
KJ Faucette ◽  
LA Fitzgerald ◽  
L Liu ◽  
CJ Parker ◽  
GM Rodgers
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Quantitative Polymerase Chain Reaction ◽  
Scavenger Receptor ◽  
Kinase C ◽  
Normal Human ◽  
Membrane Bound ◽  
Polymerase Chain ◽  
Umbilical Vein Endothelial Cells

Normal human plasma contains procoagulant albumin (PC-Al), an anionic form of albumin that induces tissue factor (TF) activity in human umbilical vein endothelial cells (HUVEC) and monocytes. In this study, we investigated both the interactions between HUVEC and PC-Al and the mechanism by which PC-Al induces TF activity. Binding of PC-Al to HUVEC was specific and reversible. Further studies indicated that membrane- bound PC-Al was not internalized by HUVEC. A potential receptor on HUVEC was suggested by studies in which the capacity of a variety of reagents to inhibit the activity of PC-Al was quantitated. Induction of TF activity by PC-Al was antagonized by dextran sulfate, heparin, fucoidan, and concanavalin A but not by ovalbumin, polyglutamic acid, or polyvinyl sulfate. This competition profile bears similarities to those reported for scavenger receptors that have been identified on both HUVEC and monocytes. Involvement of protein kinase C (PKC) in the PC-Al-induced enhancement of TF activity was suggested by experiments in which staurosporine, an inhibitor of PKC, suppressed the activity of PC-Al. The induction of TF activity by PC-Al was further characterized by using a quantitative polymerase chain reaction assay. Increased TF mRNA was first seen after 1 hour of incubation with PC-Al. Maximal observed expression occurred at 2 hours, but at 5 hours, expression had significantly decreased. Monocytes could also be induced to express TF mRNA after a 2-hour incubation with PC-Al. These results suggest that the functionally relevant binding of PC-Al to HUVEC may be mediated through interactions with a membrane constituent that has some of the properties of a scavenger receptor and that this interaction augments TF activity by enhancing transcription of TF mRNA, at least in part, by a mechanism that is dependent on activation of PKC.

scholarly journals Characterization of the interactions between procoagulant albumin and human endothelial cells

Blood ◽  
1993 ◽  
Vol 82 (9) ◽  
pp. 2684-2692 ◽  
Author(s):  
KJ Faucette ◽  
LA Fitzgerald ◽  
L Liu ◽  
CJ Parker ◽  
GM Rodgers
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Quantitative Polymerase Chain Reaction ◽  
Scavenger Receptor ◽  
Kinase C ◽  
Normal Human ◽  
Membrane Bound ◽  
Polymerase Chain ◽  
Umbilical Vein Endothelial Cells

Abstract Normal human plasma contains procoagulant albumin (PC-Al), an anionic form of albumin that induces tissue factor (TF) activity in human umbilical vein endothelial cells (HUVEC) and monocytes. In this study, we investigated both the interactions between HUVEC and PC-Al and the mechanism by which PC-Al induces TF activity. Binding of PC-Al to HUVEC was specific and reversible. Further studies indicated that membrane- bound PC-Al was not internalized by HUVEC. A potential receptor on HUVEC was suggested by studies in which the capacity of a variety of reagents to inhibit the activity of PC-Al was quantitated. Induction of TF activity by PC-Al was antagonized by dextran sulfate, heparin, fucoidan, and concanavalin A but not by ovalbumin, polyglutamic acid, or polyvinyl sulfate. This competition profile bears similarities to those reported for scavenger receptors that have been identified on both HUVEC and monocytes. Involvement of protein kinase C (PKC) in the PC-Al-induced enhancement of TF activity was suggested by experiments in which staurosporine, an inhibitor of PKC, suppressed the activity of PC-Al. The induction of TF activity by PC-Al was further characterized by using a quantitative polymerase chain reaction assay. Increased TF mRNA was first seen after 1 hour of incubation with PC-Al. Maximal observed expression occurred at 2 hours, but at 5 hours, expression had significantly decreased. Monocytes could also be induced to express TF mRNA after a 2-hour incubation with PC-Al. These results suggest that the functionally relevant binding of PC-Al to HUVEC may be mediated through interactions with a membrane constituent that has some of the properties of a scavenger receptor and that this interaction augments TF activity by enhancing transcription of TF mRNA, at least in part, by a mechanism that is dependent on activation of PKC.

Partial Characterization of a Fibrinolytic Inhibitor Produced by Cultured Endothelial Cells Derived from Human Umbilical Vein

1982 ◽  
Vol 47 (02) ◽  
pp. 128-131 ◽  
Author(s):  
F Esnard ◽  
E Dupuy ◽  
A M Dosne ◽  
E Bodevin
Keyword(s):  
Endothelial Cells ◽  
Primary Culture ◽  
Ammonium Sulphate ◽  
Concanavalin A ◽  
Umbilical Vein ◽  
Human Umbilical Vein ◽  
Preliminary Characterization ◽  
Umbilical Vein Endothelial Cells ◽  
Ammonium Sulphate Saturation

SummaryA preliminary characterization of a fibrinolytic inhibitor released by human umbilical vein endothelial cells in primary culture is reported. This molecule of Mr comprised between 2 × 105 and 106 and of μ2 mobility precipitates at 43% ammonium sulphate saturation and is totally adsorbed on Concanavalin A Sepharose 4 B. A possible relationship with a macroglobulins is discussed.

Abstract 1746: Insulin Like Growth Factor Binding Protein-3 (IGFBP-3) Mediates Vascular Repair By HDL Receptor Activation And Sphingosine Kinase (SK-1)/Sphingosine 1-Phosphate (S1P) Dependent Nitric Oxide (NO) Generation

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Sergio Li Calzi ◽  
Jennifer L Kielczewski ◽  
Evan McFarland ◽  
Kyung Hee Chang ◽  
Aqeela Afzal ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Scavenger Receptor ◽  
Receptor Activation ◽  
Hematopoietic Stem ◽  
Beneficial Effects ◽  
Sphingosine 1 Phosphate ◽  
Active Motor ◽  
Umbilical Vein Endothelial Cells ◽  
Igfbp 3

Purpose: We demonstrated that IGFBP-3 stimulates hematopoietic stem cells (HSC) to differentiate into endothelial cells, form capillaries, and stabilize the vasculature (Chang, et al, PNAS 2007). Local IGFBP- 3 production is increased by hypoxia and facilitates the homing of HSC to areas of injury. In the circulation, IGFBP-3 is bound to HDL. In this study, we investigated the signaling pathways responsible for the robust migratory effects of IGFBP-3. Methods: The effects of IGFBP-3 on NO generation in human vascular precursors (CD 34 + , CD14 − ), human lung microvascular endothelial cells, and human umbilical vein endothelial cells were examined using DAF-FM fluorescence. Western analysis was use for detection of eNOS and vasodilator-stimulated phosphoprotein (VASP), which redistributes to lamellipodia forming an active motor complex that supports motility and is phosphorylated in response to NO. Localization of VASP was performed by immunohistochemistry. SK-1 was assessed following IGFBP-3 stimulation. Results: In CD34 + cells and endothelial cells, IGFBP-3 stimulated eNOS phosphorylation at Ser1177 (102 ± 1.8%, P = 0.0002) and increased NO generation (275 ± 50%, P = 0.05) by increasing SK-1 and S1P generation. IGFBP-3 was bound and internalized by the HDL receptor, scavenger receptor 1B (SR1B). NO generation following IGFBP-3 exposure was reduced by SK inhibitors or SR-1B blocking antibody pretreatment (35 ± 5%, P < 0.02). IGFBP-3 generated NO increased phosphorylation of VASP at Ser239 and promoted the redistribution of VASP to lamellipodia. Conclusions: IGFBP-3 effects on cell migration are NO dependent and mediated in part by activation of the HDL receptor SR1B suggesting that some of the beneficial effects of HDL are mediated by the association of IGFBP-3.

scholarly journals Omentin-1 Ameliorated Free Fatty Acid-Induced Impairment in Proliferation, Migration, and Inflammatory States of HUVECs

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Yubin Chen ◽  
Fen Liu ◽  
Fei Han ◽  
Lizhi Lv ◽  
Can-e Tang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Fatty Acid ◽  
Endothelial Cell ◽  
Free Fatty Acid ◽  
Cell Injury ◽  
Umbilical Vein ◽  
Quantitative Polymerase Chain Reaction ◽  
Cell Counting ◽  
Endothelial Cell Injury ◽  
Umbilical Vein Endothelial Cells

Objectives. Endothelial cell injury is a critical pathological change during the development of atherosclerosis. Here, we explored the effect of omentin-1 on free fatty acid- (FFA-) induced endothelial cell injury. Methods. An FFA-induced endothelial cell injury model was established to investigate the role of omentin-1 in this process. Cell proliferation was analyzed with the Cell Counting Kit assay and flow cytometry. Scratch and transwell assays were used to evaluate cell migration. Factors secreted by endothelial cells after injury were detected by western blotting, reverse-transcription quantitative polymerase chain reaction, and cellular fluorescence assay. Results. Omentin-1 rescued the FFA-induced impaired proliferation and migration capabilities of human umbilical vein endothelial cells (HUVECs). It decreased the number of THP-1 cells attached to HUVECs in response to injury and inhibited the FFA-induced proinflammatory state of HUVECs. Conclusion. Omentin-1 could partly ameliorate FFA-induced endothelial cell injury.

Cyclical mechanical stretch enhances angiopoietin-2 and Tie2 receptor expression in cultured human umbilical vein endothelial cells

2003 ◽  
Vol 104 (4) ◽  
pp. 421-428 ◽  
Author(s):  
Hang CHANG ◽  
Bao-Wei WANG ◽  
Peiliang KUAN ◽  
Kou-Gi SHYU
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Mechanical Stretch ◽  
Receptor Proteins ◽  
Herbimycin A ◽  
Tie2 Receptor ◽  
Kinase C ◽  
Umbilical Vein Endothelial Cells ◽  
Angiopoietin 2 ◽  
Angiopoietin 1

Endothelial cells are essential for neovascularization. Angiopoietins and Tie receptors are required for a normal vasculature. How cyclical mechanical stretch affects the expression of components of the angiopoietin system is not known. In this study, we investigated the regulation of angiopoietins and Tie receptors by cyclical mechanical stretch in cultured human umbilical vein endothelial cells (HUVECs). HUVECs grown on a flexible membrane base were stretched by vacuum to 20% elongation, at 60cycles/min. The levels of angiopoietin-2 protein began to increase as early as 2h after stretch was initially applied, reached a maximum of 2.7-fold over the control value by 6h. The Tie2 receptor protein showed the same pattern as Ang-2. These increases in angiopoietin-2 and Tie2 receptor proteins at 6h were blocked by the addition (30min before stretch) of the protein kinase C inhibitor Gö6976 (16nM) or the tyrosine kinase inhibitor herbimycin A (24µM). Similar to protein expression, the levels of angiopoietin-2 and Tie2 receptor mRNAs in HUVECs increased 3.1-fold and 2.5-fold respectively after stretch for 6h. These increases were also blocked by Gö6976 or herbimycin A. Cyclical mechanical stretch increased (and Gö6976 or herbimycin A abrogated these increases) the immunohistochemical labelling of angiopoietin-2 and Tie2 receptor after a 6h stretch. The levels of angiopoietin-1 and Tie1 receptor proteins, mRNAs and immunohistochemical staining were unaffected by cyclical mechanical stretch. Thus cyclical mechanical stretch activates the expression of angiopoietin-2 and the Tie2 receptor, but not angiopoietin-1 or the Tie1 receptor, in cultured HUVECs. This mechanical effect is probably mediated by the tyrosine kinase and protein kinase C pathways.

scholarly journals Down-regulated miR-145 rescues insulin resistance in endothelial cells

2019 ◽  
Author(s):  
Jing Wang ◽  
Zhichun Dong ◽  
Liying Lou
Keyword(s):  
Insulin Resistance ◽  
Endothelial Cells ◽  
High Glucose ◽  
Umbilical Vein ◽  
Glucose Consumption ◽  
Over Expression ◽  
Qrt Pcr ◽  
Starting Point ◽  
Polymerase Chain ◽  
Umbilical Vein Endothelial Cells

Abstract Background: MiR-145 is involved in insulin resistance (IR) in liver cells, but its effects in human umbilical vein endothelial cells (HUVECs) induced by IR remains unclear. This study took this as the starting point, aiming to find a potential target for the treatment of related disease. Methods: HUVECs were respectively treated with glucose of 15, 30, 45 mmol/L, or insulin of 1, 2, 3, 4, 5 μmol/L on the basis of high-glucose (33.3 mmol/L). MiR-145 mimics and miR-145 inhibitor were severally transfected into HUVECs with or without IR (4 μmol/L insulin + high-glucose). Quantitative real-time polymerase chain reaction (qRT-PCR) assay determined the miR-145 expression in HUVECs after treatment and transfection. The glucose consumption and glycogen contents of cells were appraised by glucose oxidase-peroxidase and anthranone-sulfuric acid methods, respectively. The apoptotic rates were ascertained using the flow cytometry. The expressions of apoptosis-related indicators Bcl-2 and Bax were analyzed by western blot (WB) and qRT-PCR assays. Results: The expression of miR-145 was increased in IR models and incremental glucose concentrations. The glucose consumption and glycogen content were down-regulated in IR-induced HUVECs, which were enhanced by over-expressed miR-145 but reversed by down-regulation. Moreover, over-expression of miR-145 aggravated the apoptosis of IR-induced HUVECs, while the inhibition of miR-145 had a completely opposite effect. Accordingly, up-regulated miR-145 obviously reduced Bcl-2 level and enhanced Bax expression in IR models, which was contrary to the down-regulated miR-145. Conclusion: Down-regulated miR-145 rescued IR in endothelial cells, which might be a conceivable treatment for IR of endothelial cells.

Induction of Tissue Factor Synthesis in Human Umbilical Vein Endothelial Cells Involves Protein Kinase C

1992 ◽  
Vol 67 (04) ◽  
pp. 473-477 ◽  
Author(s):  
Kjell Sverre Pettersen ◽  
Merete Thune Wiiger ◽  
Nobuhiro Narahara ◽  
Kiyoshi Andoh ◽  
Gustav Gaudernack ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Tissue Factor ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Interleukin 1Β ◽  
Human Umbilical Vein ◽  
Kinase C ◽  
Umbilical Vein Endothelial Cells

SummaryIncubation of human umbilical vein endothelial cells with one of the following compounds: endotoxin, recombinant interleukin-1β, recombinant tumor necrosis factor α, allogenic lymphocyte subpopulations or phorbol ester resulted in significant induction of tissue factor synthesis. Diacylglycerol had the same effect and also enhanced synergistically the induction caused by endotoxin and interleukin-1β. Two different inhibitors of protein kinase C, H7 and sphingosine, inhibited tissue factor synthesis at concentrations which did not depress protein synthesis in general, suggesting that protein kinase C is involved in the processes leading to tissue factor synthesis. Cells down-regulated for the tissue factor response to TPA responded essentially normally to endotoxin and interleukin-1 with regard to tissue factor synthesis.

scholarly journals Biogenic Aspergillus tubingensis silver nanoparticles’ in vitro effects on human umbilical vein endothelial cells, normal human fibroblasts, HEPG2, and Galleria mellonella

2019 ◽  
Vol 8 (6) ◽  
pp. 789-801 ◽  
Author(s):  
Cristiane Angélica Ottoni ◽  
Durvanei Augusto Maria ◽  
Priscila Jane Romano de Oliveira Gonçalves ◽  
Welington Luiz de Araújo ◽  
Ana Olívia de Souza
Keyword(s):  
Silver Nanoparticles ◽  
Endothelial Cells ◽  
Galleria Mellonella ◽  
Umbilical Vein ◽  
Human Fibroblasts ◽  
Normal Fibroblast ◽  
Aspergillus Tubingensis ◽  
Human Umbilical Vein ◽  
Normal Human ◽  
Umbilical Vein Endothelial Cells

Abstract Silver nanoparticles (AgNPs) are widely incorporated into different hygiene, personal care, and healthcare products. However, few studies have been undertaken to determine the effects of biogenic AgNPs on human health. The effect of biosynthesized AgNPs using the fungus Aspergillus tubingensis culture was evaluated on human umbilical vein endothelial cells (HUVECs), normal human fibroblasts (FN1), human hepatoma cells (HEPG2) and a Galleria mellonella model. HUVECs were more susceptible to biogenic AgNPs than normal fibroblasts FN1 and intense cytotoxicity was observed only for very high concentrations at and above 2.5 μM for both cells. Normal human fibroblasts FN1 exposed to AgNPs for 24 h showed viability of 98.83 ± 8.40% and 94.86 ± 5.50% for 1.25 and 2.5 μM, respectively. At 5 and 10 μM, related to the control, an increase in cell viability was observed being 112.66 ± 9.94% and 117.86 ± 8.86%, respectively. Similar results were obtained for treatment for 48 and 72 h. At 1.25, 2.5, 5 and 10 μM of AgNPs, at 24 h, HUVECs showed 51.34 ± 7.47%, 27.01 ± 5.77%, 26.00 ± 3.03% and 27.64 ± 5.85% of viability, respectively. No alteration in cell distribution among different cycle phases was observed after HUVEC and normal fibroblast FN1 exposure to AgNPs from 0.01 to 1 μM for 24, 48 and 72 h. Based on the clonogenic assay, nanoparticles successfully inhibited HEPG2 cell proliferation when exposed to concentrations up to 1 μM. In addition to that, AgNPs did not induce senescence and no morphological alteration was observed by scanning electron microscopy on the endothelial cells. In the larvae of the wax moth, Galleria mellonella, a model for toxicity, AgNPs showed no significant effects, which corroborates to the safety of their use in mammalian cells. These results demonstrate that the use of A. tubingensis AgNPs is a promising biotechnological approach and these AgNPs can be applied in several biomedical situations.

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