scholarly journals Rapid Internalization and Nuclear Translocation of CCL5 and CXCL4 in Endothelial Cells

2021 ◽  
Vol 22 (14) ◽  
pp. 7332
Author(s):  
Annemiek Dickhout ◽  
Dawid M. Kaczor ◽  
Alexandra C. A. Heinzmann ◽  
Sanne L. N. Brouns ◽  
Johan W. M. Heemskerk ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Intracellular Calcium ◽  
Nuclear Translocation ◽  
Umbilical Vein ◽  
G Protein Coupled Receptors ◽  
Cell Organelle ◽  
Intracellular Calcium Signaling ◽  
Ea.Hy926 Cells ◽  
Umbilical Vein Endothelial Cells ◽  
G Protein Coupled

The chemokines CCL5 and CXCL4 are deposited by platelets onto endothelial cells, inducing monocyte arrest. Here, the fate of CCL5 and CXCL4 after endothelial deposition was investigated. Human umbilical vein endothelial cells (HUVECs) and EA.hy926 cells were incubated with CCL5 or CXCL4 for up to 120 min, and chemokine uptake was analyzed by microscopy and by ELISA. Intracellular calcium signaling was visualized upon chemokine treatment, and monocyte arrest was evaluated under laminar flow. Whereas CXCL4 remained partly on the cell surface, all of the CCL5 was internalized into endothelial cells. Endocytosis of CCL5 and CXCL4 was shown as a rapid and active process that primarily depended on dynamin, clathrin, and G protein-coupled receptors (GPCRs), but not on surface proteoglycans. Intracellular calcium signals were increased after chemokine treatment. Confocal microscopy and ELISA measurements in cell organelle fractions indicated that both chemokines accumulated in the nucleus. Internalization did not affect leukocyte arrest, as pretreatment of chemokines and subsequent washing did not alter monocyte adhesion to endothelial cells. Endothelial cells rapidly and actively internalize CCL5 and CXCL4 by clathrin and dynamin-dependent endocytosis, where the chemokines appear to be directed to the nucleus. These findings expand our knowledge of how chemokines attract leukocytes to sites of inflammation.

The functional thrombin receptor is associated with the plasmalemma and a large endosomal network in cultured human umbilical vein endothelial cells

1995 ◽  
Vol 108 (3) ◽  
pp. 1155-1164 ◽  
Author(s):  
R. Horvat ◽  
G.E. Palade
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Morphological Evidence ◽  
Thrombin Receptor ◽  
Human Umbilical Vein ◽  
Cultured Endothelial Cells ◽  
Human Thrombin ◽  
Membrane Spanning ◽  
Umbilical Vein Endothelial Cells ◽  
G Protein Coupled

The functional thrombin receptor, normally expressed by endothelial cells and platelets, is a member of the G protein-coupled, seven membrane-spanning-domain receptor family and is thought to be responsible for most, if not all, the cell stimulatory effects of thrombin. Upon binding, thrombin cleaves the receptor's N-terminal ectodomain, unmasking a new N terminus, which by itself activates the receptor. Using antibodies to different domains of the human thrombin receptor, we have localized the receptor in cultured human umbilical vein endothelial cells by indirect immunofluorescence and immunoelectron microscopy. We found the receptor expressed on the plasmalemma of cultured endothelial cells in individual units rather than in clusters, at lower concentration than, and at different sites from, thrombomodulin. We also found the receptor associated with a distinct, intracellular, transferrin receptor-containing, tubulovesicular network. The thrombin receptor-positive structure spread from the perinuclear region to the periphery of the cells, exhibiting a number of varicosities interconnected by branching tubular elements, strikingly similar to an image recently described for a continuous endosomal reticulum. Our results provide morphological evidence for the presence of the functional thrombin receptor at relative low density on the surface of cultured endothelial cells (compared to thrombomodulin) and in relatively large quantities inside the cells, associated with an endosomal compartment.

scholarly journals Ellagic acid inhibits IL-1β-induced cell adhesion molecule expression in human umbilical vein endothelial cells

2007 ◽  
Vol 97 (4) ◽  
pp. 692-698 ◽  
Author(s):  
Ya-Mei Yu ◽  
Zhi-Hong Wang ◽  
Chung-Hsien Liu ◽  
Chin-Seng Chen
Keyword(s):  
Reactive Oxygen Species ◽  
Endothelial Cells ◽  
Cell Adhesion ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Nuclear Translocation ◽  
Umbilical Vein ◽  
Oxygen Species ◽  
Adhesion Molecule Expression ◽  
Umbilical Vein Endothelial Cells

Expression of cell adhesion molecules by endothelium and the attachment of monocytes to endothelium may play a major role in atherosclerosis. Ellagic acid (EA) is a phenolic compound found in fruits and nuts including raspberries, strawberries, grapes and walnuts. Previous studies have indicated that EA possesses antioxidant activity in vitro. In the present study, we investigated the effects of EA on the formation of intracellular reactive oxygen species, the translocation of NFκB and expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 and endothelial leucocyte adhesion molecule (E-selectin) induced by IL-1β in human umbilical vein endothelial cells (HUVEC). We found that EA significantly reduced the binding of human monocytic cell line, U937, to IL-1β-treated HUVEC. The production of reactive oxygen species by IL-1β was dose-dependently suppressed by EA. Supplementation with increasing doses of EA up to 50 μmol/l was most effective in inhibiting the expression of VCAM-1 and E-selectin. Furthermore, the inhibition of IL-1β-induced adhesion molecule expression by EA was manifested by the suppression of nuclear translocation of p65 and p50. In conclusion, EA inhibits IL-1β-induced nuclear translocation of p65 and p50, thereby suppressing the expression of VCAM-1 and E-selectin, resulting in decreased monocyte adhesion. Thus, EA has anti-inflammatory properties and may play an important role in the prevention of atherosclerosis.

scholarly journals Transcription of the sarcoplasmic/endoplasmic reticulum Ca2+-ATPase type 3 gene, ATP2A3, is regulated by the calcineurin/NFAT pathway in endothelial cells

2006 ◽  
Vol 394 (1) ◽  
pp. 27-33 ◽  
Author(s):  
Lahouaria Hadri ◽  
Catherine Pavoine ◽  
Larissa Lipskaia ◽  
Sabrina Yacoubi ◽  
Anne-Marie Lompré
Keyword(s):  
Endothelial Cells ◽  
Endoplasmic Reticulum ◽  
Nuclear Translocation ◽  
Umbilical Vein ◽  
Nuclear Factor ◽  
H1 Receptor ◽  
Activated T Cells ◽  
Cycling Treatment ◽  
Ea.Hy926 Cells ◽  
Type 3

Histamine, known to induce Ca2+ oscillations in endothelial cells, was used to alter Ca2+ cycling. Treatment of HUVEC (human umbilical-vein endothelial cell)-derived EA.hy926 cells with histamine for 1–3 days increased the levels of SERCA (sarcoplasmic/endoplasmic reticulum Ca2+-ATPase) 3, but not of SERCA 2b, transcripts and proteins. Promoter-reporter gene assays demonstrated that this increase in expression was due to activation of SERCA 3 gene transcription. The effect of histamine was abolished by mepyramine, but not by cimetidine, indicating that the H1 receptor, but not the H2 receptor, was involved. The histamine-induced up-regulation of SERCA 3 was abolished by cyclosporin A and by VIVIT, a peptide that prevents calcineurin and NFAT (nuclear factor of activated T-cells) from interacting, indicating involvement of the calcineurin/NFAT pathway. Histamine also induced the nuclear translocation of NFAT. NFAT did not directly bind to the SERCA 3 promoter, but activated Ets-1 (E twenty-six-1), which drives the expression of the SERCA 3 gene. Finally, cells treated with histamine and loaded with fura 2 exhibited an improved capacity in eliminating high cytosolic Ca2+ concentrations, in accordance with an increase in activity of a low-affinity Ca2+-ATPase, like SERCA 3. Thus chronic treatment of endothelial cells with histamine up-regulates SERCA 3 transcription. The effect of histamine is mediated by the H1R (histamine 1 receptor) and involves activation of the calcineurin/NFAT pathway. By increasing the rate of Ca2+ sequestration, up-regulation of SERCA 3 counteracts the cytosolic increase in Ca2+ concentration.

Protease-Activated Receptors Upregulate Cyclooxygenase-2 Expression in Human Endothelial Cells

2002 ◽  
Vol 88 (08) ◽  
pp. 321-328 ◽  
Author(s):  
Rebecca Houliston ◽  
Rosemary Keogh ◽  
David Sugden ◽  
Jayesh Dudhia ◽  
Tom Carter ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Mrna Expression ◽  
Umbilical Vein ◽  
Pcr Analysis ◽  
Protease Activated Receptors ◽  
Prostanoid Production ◽  
Protease Activated Receptor 1 ◽  
Cox 2 ◽  
Umbilical Vein Endothelial Cells ◽  
G Protein Coupled

SummaryWe have previously shown that the serine protease thrombin and other G protein-coupled agonists acutely enhance synthesis and release of prostacyclin from human umbilical vein endothelial cells (HUVEC) through activation of cPLA2α. Here, we show that thrombin and other physiological endothelial cell agonists upregulate COX-2 induction in HUVEC. Thrombin treatment caused a rapid and sustained increase in prostacyclin (PGI2) synthesis from HUVEC. Thrombin and a selective protease-activated receptor-1 (PAR-1) peptide (TRAP) evoked doseand time-dependent increases in COX-2 protein expression which were equivalent to that induced by the proinflammatory cytokine IL-1α. Quantitative and real-time PCR analysis showed enhanced COX-2 mRNA expression in thrombinor TRAP-stimulated HUVEC whereas COX-1 expression was unaffected. A PAR-2 agonist peptide also induced COX-2 protein and mRNA expression with kinetics distinct from those of thrombin, and promoted PGI2 release. These results demonstrate that regulation of COX-2 induction is an important functional response of HUVEC to PAR activation and suggest that PARs promote sustained upregulation of prostanoid production in human endothelium.

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