scholarly journals Uptake of polyamines by human endothelial cells. Characterization and lack of effect of agonists of endothelial function

1992 ◽  
Vol 286 (2) ◽  
pp. 413-417 ◽  
Author(s):  
D M L Morgan
Keyword(s):  
Endothelial Cells ◽  
Endothelial Function ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Therapeutic Agents ◽  
Human Endothelial Cells ◽  
Polyamine Analogues ◽  
Pharmacological Significance ◽  
Polyamine Uptake ◽  
Umbilical Vein Endothelial Cells

Uptake of polyamines by confluent monolayers of human umbilical-vein endothelial cells (HUVECs) was found to be time-, temperature- and concentration-dependent, energy-requiring, and saturable. Kinetic constants were putrescine Kt 3 +/- 1 microM, Vmax. 15 +/- 7 pmol/h per microgram of protein; spermidine, 0.7 +/- 0.2, 12 +/- 3; spermine, 1 +/- 0.7, 11 +/- 4. Putrescine uptake was inhibited by spermine or spermidine, whereas uptake of spermine or spermidine was not inhibited by 20 microM-putrescine. These data suggest the existence of two carriers, one shared by spermine and spermidine, and one capable of transporting all three polyamines. Pretreatment of HUVECs with thrombin (less than or equal to 10 units/ml; 1 h), bradykinin (less than or equal to 10 microM; 1 h), interleukin-1 (less than or equal to 100 units/ml; 2 h) or phorbol 12-myristate 13-acetate (less than or equal to 1.0 microM; 1 h), all known agonists of endothelial function, had no significant effect on polyamine uptake. These responses may be of importance in angiogenesis and wound healing, and could have pharmacological significance, for there is a growing interest in the use of polyamines or polyamine analogues as therapeutic agents.

Bacterial adherence to human endothelial cells

1988 ◽  
Vol 65 (3) ◽  
pp. 1372-1376 ◽  
Author(s):  
P. D. Thomas ◽  
F. W. Hampson ◽  
G. W. Hunninghake
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Bacterial Adherence ◽  
Pulmonary Infections ◽  
Human Endothelial Cells ◽  
Cell Surface Antigens ◽  
Lung Endothelium ◽  
Umbilical Vein Endothelial Cells ◽  
Reduced Temperature

The adult respiratory distress syndrome (ARDS) is frequently caused by exposure of the lung endothelium to circulating endotoxin (lipopolysaccharide, LPS) and pulmonary infections frequently develop during the course of ARDS. The present studies demonstrate that LPS and interleukin 1 (IL-1, a mediator released by endothelial cells after exposure to LPS) enhance the adherence of Staphylococcus aureus to human umbilical vein endothelial cells. gamma-Interferon, another mediator that induces expression of some cell surface antigens on endothelial cells, had no effect on bacterial adherence. The adherence of bacteria to endothelium was increased by prior opsonization of the bacteria with fresh human serum and was reduced by prior absorption of the serum with bacteria before the use of the serum for opsonization. The capacity of LPS to increase bacterial adherence was time dependent and was maximally expressed after 6 h of exposure; it was blocked by exposure of endothelial cells to LPS in the presence of reduced temperature or dactinomycin (Actinomycin D). These observations suggest that circulating LPS not only can trigger the development of ARDS but also may predispose the lung to the development of pulmonary infections by increasing adherence of bacteria to endothelium.

Single-Chain and Two-Chain Tissue-Type Plasminogen Activator (t-PA) Bind Differently to Cultured Human Endothelial Cells

1989 ◽  
Vol 62 (02) ◽  
pp. 699-703 ◽  
Author(s):  
Rob J Aerts ◽  
Karin Gillis ◽  
Hans Pannekoek
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Binding Sites ◽  
Umbilical Vein ◽  
Tissue Type ◽  
Single Chain ◽  
Human Endothelial Cells ◽  
Tissue Type Plasminogen Activator ◽  
Type Plasminogen Activator ◽  
Umbilical Vein Endothelial Cells

SummaryIt has recently been shown that the fibrinolytic components plasminogen and tissue-type plasminogen activator (t-PA) both bind to cultured human umbilical vein endothelial cells (HUVEC). After cleavage of t-PA by plasmin, “single-chain” t-PA (sct-PA) is converted into “two-chain” t-PA (tct-PA), which differs from the former in a number of respects. We compared binding of sct-PA and tct-PA to the surface of HUVEC. Removal of t-PA bound to HUVEC by a mild treatment with acid and a subsequent quantification of eluted t-PA both by activity- and immunoradiometric assays revealed that, at concentrations between 10 and 500 nM, HUVEC bind about 3-4 times more sct-PA than tct-PA. At these concentrations, both sct-PA and tct-PA remain active when bound to HUVEC. Mutual competition experiments showed that sct-PA and tct-PA can virtually fully inhibit binding of each other to HUVEC, but that an about twofold higher concentration of tct-PA is required to prevent halfmaximal binding of sct-PA than visa versa. These results demonstrate that sct-PA and tct-PA bind with different affinities to the same binding sites on HUVEC.

Assembly and Activation of the Intrinsic Fibrinolytic Pathway on the Surface of Human Endothelial Cells in Culture

1995 ◽  
Vol 74 (02) ◽  
pp. 698-703 ◽  
Author(s):  
Catherine Lenich ◽  
Ralph Pannell ◽  
Victor Gurewich
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Factor Xii ◽  
Plasminogen Activation ◽  
High Molecular Weight Kininogen ◽  
Human Endothelial Cells ◽  
Intrinsic Pathway ◽  
Local Fibrinolysis ◽  
Umbilical Vein Endothelial Cells ◽  
And Function

SummaryFactor XII has long been implicated in the intrinsic pathway of fibrinolysis, but the mechanism by which it triggers plasminogen activation and targets fibrinolysis has not been established. In the present study, the assembly and function of activated Factor XII (F.XIIa), prourokinase (pro-u-PA), high molecular weight kininogen (H-kininogen), and prekallikrein on human umbilical vein endothelial cells (HUVEC) was investigated. 125I-prekallikrein was shown to bind to HUVEC via receptor-bound H-kininogen in the presence of 50 μM ZnCl2. After the addition of F.XIIa, 78% of the 125I-prekallikrein initially bound to HUVEC was converted to 125I-kallikrein. However, only 6% of the HUVEC-bound 125I-pro-u-PA was thereby activated. This discrepancy was shown to be related to rapid dissociation (>50% within 15 min) of prekallikrein/kallikrein, but not pro-u-PA, from HUVEC. Increasing the level of cell-bound kallikrein increased the portion of cell-bound pro-u-PA activated, indicating that their co-localization was important for this pathway. Finally, F.XIIa was shown to trigger plasminogen activation on HUVEC via this pathway. This assembly of reactants on the endothelium suggests a mechanism whereby local fibrinolysis may be triggered by blood coagulation.

Platelets Stimulate Thromboplastin Synthesis in Human Endothelial Cells

1983 ◽  
Vol 49 (02) ◽  
pp. 069-072 ◽  
Author(s):  
U L H Johnsen ◽  
T Lyberg ◽  
K S Galdal ◽  
H Prydz
Keyword(s):  
Endothelial Cells ◽  
De Novo ◽  
Umbilical Vein ◽  
Time Dependent ◽  
De Novo Synthesis ◽  
Human Endothelial Cells ◽  
Tumor Promotor ◽  
Coagulation Assay ◽  
Platelet Aggregates ◽  
Umbilical Vein Endothelial Cells

SummaryHuman umbilical vein endothelial cells in culture synthesize thromboplastin upon stimulation with phytohaemagglutinin (PHA) or the tumor promotor 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The thromboplastin activity is further strongly enhanced in a time dependent reaction by the presence of gel-filtered platelets or platelet aggregates. This effect was demonstrable at platelet concentrations lower than those normally found in plasma, it may thus be of pathophysiological relevance. The thromboplastin activity increased with increasing number of platelets added. Cycloheximide inhibited the increase, suggesting that de novo synthesis of the protein component of thromboplastin, apoprotein III, is necessary.When care was taken to remove monocytes no thromboplastin activity and no apoprotein HI antigen could be demonstrated in suspensions of gel-filtered platelets, platelets aggregated with thrombin or homogenized platelets when studied with a coagulation assay and an antibody neutralization technique.

Syndecan-4-dependent signaling in the inhibition of endotoxin-induced endothelial adherence of neutrophils by antithrombin

2003 ◽  
Vol 90 (12) ◽  
pp. 1150-1157 ◽  
Author(s):  
Nicole Kaneider ◽  
Ellen Förster ◽  
Birgit Mosheimer ◽  
Daniel Sturn ◽  
Christian Wiedermann
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Human Neutrophils ◽  
Heparin Binding ◽  
Clotting Factors ◽  
Edema Formation ◽  
Granulocyte Elastase ◽  
Umbilical Vein Endothelial Cells ◽  
Static Conditions

SummaryCirculating endotoxin is elevated in sepsis and plays a role in endothelial dysfunction whereas antithrombin is decreased by virtue of its consumption during complex formation with clotting factors and by proteolytic degradation by granulocyte elastase. Dysfunction of endothelium results in enhanced leukocyte rolling and diapedesis into tissues leading to edema formation and injury. Antithrombin exerts beneficial effects on endothelial function in sepsis. A direct anti-inflammatory action of anti-thrombin in inflammatory cells is exerted via heparan sulfate proteoglycans. In this study, we investigated whether antithrom-bin affects endotoxin-induced adhesion of neutrophils to human endothelial cells in vitro and whether glycosaminoglycans are involved in its signaling. Adhesion of human neutrophils to monolayers of umbilical vein endothelial cells was tested under static conditions. Endothelial cells were pretreated with endotoxin, interleukin-1, heparinase-I, chondroitinase-ABC or anti-syndecan-4-antibody. Endotoxin and interleukin-1 increased neutrophil adherence to human umbilical vein endothelial cells which was inhibited by antithrombin. Concomitant incubation with pentasaccharide abolished this effect of antithrombin. Treatment of endothelial cells with heparinase or chondroitinase led to higher adhesion and prevented effects of antithrom-bin. With antibodies to syndecan-4, enhanced adhesion of neutrophils was observed. As studied by Western blotting, endo-toxin-induced signaling was diminished by antithrombin and the effect was reversible by chondroitinase or heparinase. From our results, we can conclude that endotoxin-induced adhesion of leukocytes to endothelium can be reversed by ligation of syndecan-4 with antithrombin´s heparin-binding site and interferences with stress response signaling events in endothelium.

Isolation and sequencing of a cDNA clone homologous to the v-sis oncogene from human endothelial cells

1986 ◽  
Vol 6 (8) ◽  
pp. 3018-3022
Author(s):  
B D Tong ◽  
S E Levine ◽  
M Jaye ◽  
G Ricca ◽  
W Drohan ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Growth Factor ◽  
Cdna Library ◽  
Cdna Clone ◽  
Umbilical Vein ◽  
Platelet Derived Growth Factor ◽  
Human Endothelial Cells ◽  
A Chain ◽  
Umbilical Vein Endothelial Cells ◽  
Reading Frames

A clone containing the 3' end of the mRNA for the human c-sis gene (homologous to the B chain of platelet-derived growth factor) was isolated from a cDNA library derived from human umbilical vein endothelial cells and then sequenced. The analysis of possible translation products in all three reading frames indicated that the A chain of platelet-derived growth factor was not coded for within the 3' end of the c-sis mRNA. The 3' end of the mRNA for c-sis is contained in or adjacent to exon 6.

scholarly journals Primary cilia of human endothelial cells disassemble under laminar shear stress

2004 ◽  
Vol 164 (6) ◽  
pp. 811-817 ◽  
Author(s):  
Carlo Iomini ◽  
Karla Tejada ◽  
Wenjun Mo ◽  
Heikki Vaananen ◽  
Gianni Piperno
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Primary Cilia ◽  
Umbilical Vein ◽  
Intraflagellar Transport ◽  
Mechanical Stimuli ◽  
Laminar Shear Stress ◽  
Human Endothelial Cells ◽  
Umbilical Vein Endothelial Cells ◽  
Laminar Shear

We identified primary cilia and centrosomes in cultured human umbilical vein endothelial cells (HUVEC) by antibodies to acetyl-α-tubulin and capillary morphogenesis gene-1 product (CMG-1), a human homologue of the intraflagellar transport (IFT) protein IFT-71 in Chlamydomonas. CMG-1 was present in particles along primary cilia of HUVEC at interphase and around the oldest basal body/centriole at interphase and mitosis. To study the response of primary cilia and centrosomes to mechanical stimuli, we exposed cultured HUVEC to laminar shear stress (LSS). Under LSS, all primary cilia disassembled, and centrosomes were deprived of CMG-1. We conclude that the exposure to LSS ends the IFT in cultured endothelial cells.

scholarly journals MicroRNA as Crucial Regulators of Gene Expression in Estradiol-Treated Human Endothelial Cells

2018 ◽  
Vol 45 (5) ◽  
pp. 1878-1892 ◽  
Author(s):  
Xavier Vidal-Gómez ◽  
Daniel Pérez-Cremades ◽  
Ana Mompeón ◽  
Ana Paula Dantas ◽  
Susana Novella ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Function ◽  
Pathway Analysis ◽  
Mirna Expression ◽  
Umbilical Vein ◽  
Mirna Gene ◽  
Mirna Profile ◽  
Human Endothelial Cells ◽  
Estrogen Regulation

Background/Aims: Estrogen signalling plays an important role in vascular biology as it modulates vasoactive and metabolic pathways in endothelial cells. Growing evidence has also established microRNA (miRNA) as key regulators of endothelial function. Nonetheless, the role of estrogen regulation on miRNA profile in endothelial cells is poorly understood. In this study, we aimed to determine how estrogen modulates miRNA profile in human endothelial cells and to explore the role of the different estrogen receptors (ERα, ERβ and GPER) in the regulation of miRNA expression by estrogen. Methods: We used miRNA microarrays to determine global miRNA expression in human umbilical vein endothelial cells (HUVEC) exposed to a physiological concentration of estradiol (E2; 1 nmol/L) for 24 hours. miRNA-gene interactions were computationally predicted using Ingenuity Pathway Analysis and changes in miRNA levels were validated by qRT-PCR. Role of ER in the E2-induced miRNA was additionally confirmed by using specific ER agonists and antagonists. Results: miRNA array revealed that expression of 114 miRNA were significantly modified after E2 exposition. Further biological pathway analysis revealed cell death and survival, lipid metabolism, reproductive system function, as the top functions regulated by E2. We validated changes in the most significantly increased (miR-30b-5p, miR-487a-5p, miR-4710, miR-501-3p) and decreased (miR-378h and miR-1244) miRNA and the role of ER in these E2-induced miRNA was determined. Results showed that both classical, ERα and ERβ, and membrane-bound ER, GPER, differentially regulated specific miRNA. In silico analysis of validated miRNA promoters identified specific ER binding sites. Conclusion: Our findings identify differentially expressed miRNA pathways linked to E2 in human endothelial cells through ER, and provide new insights by which estrogen can modulate endothelial function.

scholarly journals Interleukin-1 Enhances the Ability of Cultured Human Umbilical Vein Endothelial Cells to Oxidize Linoleic Acid

1995 ◽  
Vol 270 (29) ◽  
pp. 17279-17286 ◽  
Author(s):  
Mercedes Camacho ◽  
Nuria Godessart ◽  
Rosa Antón ◽  
Montserrat Garca ◽  
Lus Vila
Keyword(s):  
Endothelial Cells ◽  
Linoleic Acid ◽  
Umbilical Vein ◽  
Interleukin 1 ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells
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