scholarly journals CD36 Mediates the In Vitro Inhibitory Effects of Thrombospondin-1 on Endothelial Cells

1997 ◽  
Vol 138 (3) ◽  
pp. 707-717 ◽  
Author(s):  
David W. Dawson ◽  
S. Frieda A. Pearce ◽  
Ruiqin Zhong ◽  
Roy L. Silverstein ◽  
William A. Frazier ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Fusion Proteins ◽  
Solid Phase ◽  
Umbilical Vein ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Microvascular Endothelial Cells ◽  
Thrombospondin 1 ◽  
Microvascular Endothelial

Thrombospondin-1 (TSP-1) is a naturally occurring inhibitor of angiogenesis that is able to make normal endothelial cells unresponsive to a wide variety of inducers. Here we use both native TSP-1 and small antiangiogenic peptides derived from it to show that this inhibition is mediated by CD36, a transmembrane glycoprotein found on microvascular endothelial cells. Both IgG antibodies against CD36 and glutathione-S-transferase–CD36 fusion proteins that contain the TSP-1 binding site blocked the ability of intact TSP-1 and its active peptides to inhibit the migration of cultured microvascular endothelial cells. In addition, antiangiogenic TSP-1 peptides inhibited the binding of native TSP-1 to solid phase CD36 and its fusion proteins, as well as to CD36-expressing cells. Additional molecules known to bind CD36, including the IgM anti-CD36 antibody SM∅, oxidized (but not unoxidized) low density lipoprotein, and human collagen 1, mimicked TSP-1 by inhibiting the migration of human microvascular endothelial cells. Transfection of CD36-deficient human umbilical vein endothelial cells with a CD36 expression plasmid caused them to become sensitive to TSP-1 inhibition of their migration and tube formation. This work demonstrates that endothelial CD36, previously thought to be involved only in adhesion and scavenging activities, may be essential for the inhibition of angiogenesis by thrombospondin-1.

scholarly journals Cardiac microvascular endothelial cells express α-smooth muscle actin and show low NOS III activity

1999 ◽  
Vol 276 (5) ◽  
pp. H1755-H1768 ◽  
Author(s):  
Hiroshi Ando ◽  
Thomas Kubin ◽  
Wolfgang Schaper ◽  
Jutta Schaper
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Actin ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Microvascular Endothelial Cells ◽  
Low Grade ◽  
Muscle Actin ◽  
Microvascular Endothelial

We established a culture system of porcine coronary microvascular endothelial cells (MVEC) with high cellular yield and purity >98%. Endothelial origin was confirmed by immunostaining, immunoblotting and fluorescence-activated cell sorter (FACS) analysis using low-density lipoprotein uptake, CD31, von Willebrand factor, and the lectin Dolichos biflorus agglutinin. MVEC were positive for α-smooth muscle actin in culture and in myocardium, as confirmed by FACS. Of the primary MVEC, ∼30% expressed nitric oxide synthase (NOS) III in numbers decreasing from the first passage (6 ± 1%) to the second passage (4 ± 1%; P < 0.001 vs. primary isolates), whereas ∼100% of aortic endothelial cells (AEC) expressed NOS III. In AEC, NOS III activity (pmol citrulline ⋅ mg protein−1 ⋅ min−1) was 80 ± 10 and was nearly abolished in the absence of calcium (5 ± 1, P < 0.001). In primary MVEC, however, NOS III activity in the presence and absence of calcium was 20 ± 4 and 25 ± 5, respectively. We conclude that cardiac MVEC, in contrast to AEC, contain α-smooth muscle actin, show low-grade NOS III activity, and provide a suitable in vitro system for the study of endothelial pathophysiology.

scholarly journals Thrombin Modulates the Expression of a Set of Genes Including Thrombospondin-1 in Human Microvascular Endothelial Cells

2005 ◽  
Vol 280 (23) ◽  
pp. 22172-22180 ◽  
Author(s):  
Joseph N. McLaughlin ◽  
Maria R. Mazzoni ◽  
John H. Cleator ◽  
Laurie Earls ◽  
Ana Luisa Perdigoto ◽  
...  
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Umbilical Vein ◽  
Microvascular Endothelial Cells ◽  
Microvascular Endothelium ◽  
Disease States ◽  
Similar Expression Profile ◽  
Thrombospondin 1 ◽  
Microvascular Endothelial

Thrombospondin-1 (THBS1) is a large extracellular matrix glycoprotein that affects vasculature systems such as platelet activation, angiogenesis, and wound healing. Increases in THBS1 expression have been liked to disease states including tumor progression, atherosclerosis, and arthritis. The present study focuses on the effects of thrombin activation of the G-protein-coupled, protease-activated receptor-1 (PAR-1) on THBS1 gene expression in the microvascular endothelium. Thrombin-induced changes in gene expression were characterized by microarray analysis of ∼11,000 different human genes in human microvascular endothelial cells (HMEC-1). Thrombin induced the expression of a set of at least 65 genes including THBS1. Changes in THBS1 mRNA correlated with an increase in the extracellular THBS1 protein concentration. The PAR-1-specific agonist peptide (TFLLRNK-PDK) mimicked thrombin stimulation of THBS1 expression, suggesting that thrombin signaling is through PAR-1. Further studies showed THBS1 expression was sensitive to pertussis toxin and protein kinase C inhibition indicating Gi/o- and Gq-mediated pathways. THBS1 up-regulation was also confirmed in human umbilical vein endothelial cells stimulated with thrombin. Analysis of the promoter region of THBS1 and other genes of similar expression profile identified from the microarray predicted an EBOX/EGRF transcription model. Expression of members of each family, MYC and EGR1, respectively, correlated with THBS1 expression. These results suggest thrombin formed at sites of vascular injury increases THBS1 expression into the extracellular matrix via activation of a PAR-1, Gi/o, Gq, EBOX/EGRF-signaling cascade, elucidating regulatory points that may play a role in increased THBS1 expression in disease states.

scholarly journals Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein.

1984 ◽  
Vol 99 (6) ◽  
pp. 2034-2040 ◽  
Author(s):  
J C Voyta ◽  
D P Via ◽  
C E Butterfield ◽  
B R Zetter
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Muscle Cells ◽  
Microvascular Endothelial Cells ◽  
Low Density ◽  
Fluorescent Intensity ◽  
Microvascular Endothelial

Acetylated-low density lipoprotein (Ac-LDL) is taken up by macrophages and endothelial cells via the "scavenger cell pathway" of LDL metabolism. In this report, aortic and microvascular endothelial cells internalized and degraded 7-15 times more [125I]-Ac-LDL than did smooth muscle cells or pericytes. Bound [125I]-Ac-LDL was displaced by unlabeled Ac-LDL, but not unmodified LDL. The ability to identify endothelial cells based on their increased metabolism of Ac-LDL was examined using Ac-LDL labeled with the fluorescent probe 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (Dil-Ac-LDL). When cells were incubated with 10 micrograms/ml Dil-Ac-LDL for 4 h at 37 degrees C and subsequently examined by fluorescence microscopy, capillary and aortic endothelial cells were brilliantly fluorescent whereas the fluorescent intensity of retinal pericytes and smooth muscle cells was only slightly above background levels. Dil-Ac-LDL at the concentration used for labeling cells had no effect on endothelial cell growth rate. When primary cultures of bovine adrenal capillary cells were labeled with 10 micrograms/ml of Dil-Ac-LDL for 4 h at 37 degrees C, then trypsinized and subjected to fluorescence-activated cell sorting, pure cultures of capillary endothelial cells could be obtained. Utilizing this method, large numbers of early passage microvascular endothelial cells can be obtained in significantly less time than with conventional methods.

scholarly journals Curcumin Alleviates Palmitic Acid-Induced LOX-1 Upregulation by Suppressing Endoplasmic Reticulum Stress in HUVECs

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Ruixi Luo ◽  
Lifeng Zhao ◽  
Shuaishuai Li ◽  
Peng Chen ◽  
La Wang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endoplasmic Reticulum ◽  
Palmitic Acid ◽  
Er Stress ◽  
Umbilical Vein ◽  
Low Density Lipoprotein ◽  
Critical Role ◽  
Density Lipoprotein ◽  
Low Density

Excessive free fatty acid- (FFA-) induced endothelial lipotoxicity is involved in the pathogenesis of atherosclerosis. Endoplasmic reticulum (ER) stress is mechanistically related to endothelial lipotoxicity. Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is the major oxidatively modified low-density lipoprotein (OxLDL) receptor in endothelial cells and is highly abundant in atherosclerotic lesions. Curcumin reduces the LOX-1 expression; however, the mechanism underlying this effect remains unknown. In the current study, we explored whether curcumin ameliorates palmitic acid- (PA-) induced endothelial lipotoxicity and LOX-1 upregulation by reducing ER stress in human umbilical vein endothelial cells (HUVECs). We built endothelial lipotoxicity in vitro and found that LOX-1 was upregulated after PA stimulation, during which ER stress played an important role. Next, we observed that curcumin substantially alleviated PA-induced lipotoxicity by restoring cell viability, increasing angiogenesis, and decreasing lipid deposition. Furthermore, LOX-1 upregulation in HUVECs was blocked by curcumin, possibly via ER stress suppression. Overall, our findings demonstrated that curcumin alleviates endothelial lipotoxicity and LOX-1 upregulation, and ER stress inhibition may play a critical role in this effect.

scholarly journals Human Induced Pluripotent Stem Cell-Derived Brain Endothelial Cells: Current Controversies

2021 ◽  
Vol 12 ◽  
Author(s):  
Tyler M. Lu ◽  
José Gabriel Barcia Durán ◽  
Sean Houghton ◽  
Shahin Rafii ◽  
David Redmond ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
De Novo ◽  
Density Lipoprotein ◽  
Microvascular Endothelial Cells ◽  
Brain Microvascular Endothelial Cells ◽  
Brain Endothelial Cells ◽  
Microvascular Endothelial

Brain microvascular endothelial cells (BMECs) possess unique properties that are crucial for many functions of the blood-brain-barrier (BBB) including maintenance of brain homeostasis and regulation of interactions between the brain and immune system. The generation of a pure population of putative brain microvascular endothelial cells from human pluripotent stem cell sources (iBMECs) has been described to meet the need for reliable and reproducible brain endothelial cells in vitro. Human pluripotent stem cells (hPSCs), embryonic or induced, can be differentiated into large quantities of specialized cells in order to study development and model disease. These hPSC-derived iBMECs display endothelial-like properties, such as tube formation and low-density lipoprotein uptake, high transendothelial electrical resistance (TEER), and barrier-like efflux transporter activities. Over time, the de novo generation of an organotypic endothelial cell from hPSCs has aroused controversies. This perspective article highlights the developments made in the field of hPSC derived brain endothelial cells as well as where experimental data are lacking, and what concerns have emerged since their initial description.

scholarly journals On-chip perivascular niche with patient-derived glioma cells

2020 ◽  
Author(s):  
Magda Gerigk ◽  
Harry Bulstrode ◽  
HaoTian Harvey Shi ◽  
Felix Tönisen ◽  
Camilla Cerutti ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
3D Culture ◽  
Glioma Cells ◽  
Microvascular Endothelial Cells ◽  
Normal Brain ◽  
Tumour Mass ◽  
Perivascular Niche ◽  
Microvascular Endothelial

AbstractGlioblastoma multiforme (GBM), is the most common and the most aggressive type of primary brain malignancy. Glioblastoma stem-like cells (GSCs) are able to migrate in vascular niches within or away from the tumour mass, increasing tumour resistance to patient treatments and contributing to relapses. To study individual GSCs migration and their interactions with the microenvironment in the vasculature, there is a need to develop a model of human blood vessels in vitro. Herein, we report a systematic study on the interaction between patient-derived glioma stem-like cell lines with different organotypic perivascular niche models. A microfluidic chip integrated with an extracellular matrix was fabricated to support the culture of rounded microvessels, formed with endothelial cells from three different organs, (1) human brain microvascular endothelial cells (hCMEC/D3), (2) human umbilical vein endothelial cells (HUVECs) and, (3) human lung microvascular endothelial cells (HMVEC-L). Three-dimensional (3D) cell culture retains selected adherent and tight junction markers of the endothelial cells, and the stemness-related genes of GSCs. We optimized the experimental protocol to perform qPCR, and western blot on the co-cultured GSCs with endothelial cells forming microvessels. Endpoint biological assays showed upregulation of neovascularization-related genes in endothelial cells (e.g., angiopoietins, vascular endothelial growth factor receptors) resulted after their co-culture with GBM cells. Moreover, we measured cancer cell speed and polarization during migration towards the endothelial cell formed vessel by live-cell imaging showing that organotypic (brain cancer cells – brain endothelial microvessel) interactions differ from those within non-tissue specific vascular niches. The development and optimization of this 3D microfluidic device could provide the next level of complexity of an in vitro system to study the influence of glioma cells on normal brain endothelium. More importantly, it enables the possibility to conduct comparative studies to dissect the influence of 3D culture, microvessel architecture and organotypic vessel types on glioma cells’ stemness and migration.

scholarly journals Evidence against the Hypothesis that Endothelial Endothelin B Receptor Expression Is Regulated by Relaxin and Pregnancy

Endocrinology ◽  
2005 ◽  
Vol 146 (6) ◽  
pp. 2791-2797 ◽  
Author(s):  
Laurie J. Kerchner ◽  
Jacqueline Novak ◽  
Karen Hanley-Yanez ◽  
Ketah D. Doty ◽  
Lee A. Danielson ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Renal Arteries ◽  
Receptor Expression ◽  
Receptor Protein ◽  
Microvascular Endothelial Cells ◽  
Etb Receptor ◽  
Microvascular Endothelial ◽  
Endothelin B

Abstract The endothelial endothelin B (ETB) receptor subtype is critical for renal vasodilation induced by relaxin in nonpregnant rats and during pregnancy (the latter via endogenous circulating relaxin). Here we tested whether expression of vascular ETB receptor protein is regulated by relaxin. Small renal arteries were harvested from virgin and midterm pregnant rats as well as nonpregnant rats that were administered recombinant human relaxin (rhRLX) at 4 μg/h or vehicle for 5 d or 4–6 h. Small renal arteries dissected from additional virgin rats were incubated in vitro with rhRLX or vehicle for 3 h at 37 C. ETB expression was also evaluated in cultured human endothelial cells: aortic, coronary, umbilical vein, and dermal microvascular endothelial cells. Cells were incubated for 4, 8, or 24 h with rhRLX (5, 1, or 0.1 ng/ml) or vehicle. ETB protein expression in arteries and cells was evaluated by Western analysis. No regulation of ETB expression was observed in small renal arteries in any of the experimental protocols, nor was there an increase in the vasorelaxation response to ET-3 in small renal arteries incubated in vitro with rhRLX. rhRLX only sporadically altered ETB expression in human coronary artery endothelial cells and human umbilical vein endothelial cells at certain time points or doses, and no regulation was observed in human aortic endothelial cells or human dermal microvascular endothelial cells. These results suggest that regulation of ETB receptor protein has little or no role in relaxin stimulation of the endothelial ETB/nitric oxide vasodilatory pathway.

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