Cultured endothelial cells from distinct vascular areas show differential responses to agonists

1994 ◽  
Vol 72 (9) ◽  
pp. 1007-1012 ◽  
Author(s):  
N. Woodley ◽  
J. K. Barclay
Keyword(s):  
Endothelial Cells ◽  
Nous Avons ◽  
Mots Clés ◽  
Cultured Endothelial Cells ◽  
Differential Responses

Nous avons comparé la capacité des cellules endothéliales isolées de l'aorte, de la veine cave, de la chambre ventriculaire et du système microvasculaire pulmonaire du lapin de produire un ou des facteurs de relaxation en réponse à l'acetylcholine (ACh) et à la bradykinine (BK). Des anneaux aortiques de lapin dépouillés d'endothélium ont été précontractés avec 1 μM de phényléphrine et superfusés à 2 mL/min avec une solution tampon bicarbonatée de Krebs–Henseleit. Les anneaux ont été exposées à des épreuves témoins d'embols de 3 mL de 1 μM d'ACh ou 1 μM de BK. Les embols d'ACh et de BK ont été ajoutés à des cultures de cellules endothéliales qui avaient été incubées pendant 45 min dans des milieux contenant ou non 10 μM de NG-nitro-L-arginine (NNLA). La solution ainsi obtenue a été répandue sur les anneaux en moins de 8 s. Seules les cellules endothéliales ventriculaires gauches stimulées avec ACh et BK et les cellules endothéliales microvasculaires pulmonaires stimulées avec BK ont donné des produits qui ont relaxé les anneaux d'approximativement 6 ± 2%. L'incubation avec la NNLA a atténué ces relaxations. Nos résultats indiquent qu'il existe des différences dans la capacité des cellules endothéliales de divers sites anatomiques de libérer des facteurs de relaxation dérivés de l'oxyde nitrique en réponse à l'ACh et à la BK. Mots clés : facteur de relaxation dérivé de l'endothélium, aorte, dosage biologique in vitro, acétylcholine, bradykinine, superoxyde dismutase, nitroprussiate de sodium.[Traduit par la rédaction]

Histological interaction of cultured endothelial cells and endovascular embolic materials coated with extracellular matrix

1997 ◽  
Vol 86 (1) ◽  
pp. 109-112 ◽  
Author(s):  
Shinichi Tamatani ◽  
Tsunenori Ozawa ◽  
Takashi Minakawa ◽  
Shigekazu Takeuchi ◽  
Tetsuo Koike ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Interventional Treatment ◽  
Clinical Use ◽  
Content Type ◽  
Cultured Endothelial Cells ◽  
Coated Materials ◽  
Embolic Materials ◽  
Polyvinyl Alcohol Particles

✓ This study was undertaken to evaluate the histological reaction of cultured endothelial cells to endovascular embolic materials in vitro. Endothelial cells were isolated and cultured from a canine carotid artery. Embolic materials (platinum microcoils, polyvinyl alcohol particles, silicon balloons, or silk threads), either in their normal state or after having been coated with type 1 collagen, fibronectin, or laminin, were placed on endothelial cells and cocultured for 6, 12, and 24 hours and 2, 3, 7, 14, and 21 days. The cocultures were investigated histologically using a scanning electron microscope. Endothelial cells were not found on any uncoated embolic materials, even at 21 days. On the materials coated with fibronectin or laminin, endothelial cells began to proliferate in 7 days, covering the materials extensively in 14 days. On the other hand, endothelial cells began to proliferate on the collagen-coated materials in 3 days, covering them extensively in 7 days and reaching confluence with a cobblestone pattern in 21 days. The densities of endothelial cells on collagen-coated materials were much higher than those observed on the materials coated with other extracellular matrices. Future advantages of the clinical use of collagen-coated embolic materials in interventional treatment are discussed.

A High Concentration of Melatonin Inhibits In Vitro LDL Peroxidation But Not Oxidized LDL Toxicity Toward Cultured Endothelial Cells

1998 ◽  
Vol 32 (4) ◽  
pp. 582-592 ◽  
Author(s):  
E. Walters-Laporte ◽  
C. Furman ◽  
S. Fouquet ◽  
F. Martin-Nizard ◽  
S. Lestavel ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Oxidized Ldl ◽  
High Concentration ◽  
Cultured Endothelial Cells

PGI2 is not a major prostanoid produced by cultured rabbit renal microvascular endothelial cells

1989 ◽  
Vol 256 (2) ◽  
pp. F266-F273
Author(s):  
A. Chaudhari ◽  
A. Pedram ◽  
M. A. Kirschenbaum
Keyword(s):  
Endothelial Cells ◽  
Rabbit Kidney ◽  
Angiotensin Converting Enzyme Activity ◽  
Glomerular Function ◽  
Cultured Endothelial Cells ◽  
Large Numbers ◽  
Afferent Arterioles ◽  
In Vitro Biosynthesis ◽  
Current Report

The microvasculature is known to be a source of a number of vasoregulatory prostanoids. In the kidney, these prostanoids have been proposed to influence vascular, tubular, and glomerular function. A rapid method for isolation of large numbers of preglomerular renal microvessels (interlobular arteries and afferent arterioles) from the rabbit kidney has recently been developed in this laboratory. In the current report, we describe methods to culture endothelial cells derived from these isolated renal microvessels. Endothelial cells in primary and continuous cultures were grown in monolayers on culture dishes and plates. These cells demonstrated morphology consistent with that described for other endothelial cells in culture including the presence of Weibel-Palade bodies as seen by electron microscopy. The presence of factor VIII immunofluorescence and angiotensin converting-enzyme activity was also observed. The cultured endothelial cells synthesized a number of common prostanoids under in vitro conditions and the hierarchy of biosynthesis was PGE2 greater than PGF2 alpha greater than PGI2 greater than TxA2. The ratio of the in vitro biosynthesis of PGI2:PGE2 was approximately 1:5, as compared with a 3-5:1 ratio seen in freshly isolated intact microvessels. Prostanoid biosynthesis increased in the cultured endothelial cells in the presence of arachidonic acid (1 and 10 microM), A23187 (10 microM), thrombin (5 U/ml), or bradykinin (1 microM) and decreased with mepacrine (10 microM)-or indomethacin (100 microM), suggesting that these cells were metabolically responsive to a variety of prostanoid stimulators and inhibitors. In summary, endothelial cells can be cultured from freshly isolated preglomerular renal microvessels and have the ability to produce a number of vasoregulatory prostanoids under in vitro conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Deleterious effects of endotoxin on cultured endothelial cells: An in vitro model of vascular injury

Inflammation ◽  
1981 ◽  
Vol 5 (2) ◽  
pp. 115-126 ◽  
Author(s):  
Osamu Yamada ◽  
Charles F. Moldow ◽  
Thomas Sacks ◽  
Philip R. Craddock ◽  
M. A. Boogaerts ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Vascular Injury ◽  
In Vitro Model ◽  
Cultured Endothelial Cells ◽  
Vitro Model

Glucose Inhibits Prostacyclin Production by Cultured Aortic Endothelial Cells

1988 ◽  
Vol 60 (02) ◽  
pp. 174-177 ◽  
Author(s):  
Hiroshi Ono ◽  
Fumio Umeda ◽  
Toyoshi Inoguchi ◽  
Hiroshi Ibayashi
Keyword(s):  
Endothelial Cells ◽  
Vascular Wall ◽  
Diabetic Patients ◽  
High Concentration ◽  
Aortic Endothelial Cells ◽  
Cultured Endothelial Cells ◽  
Effect Of Glucose ◽  
Prostacyclin Production ◽  
Aortic Endothelial

SummaryA reduction in production of prostacyclin (PGI2) by the cells in the vascular wall may play a role in the pathogenesis of atherosclerosis in diabetic patients. The present study was undertaken to evaluate the effect of glucose on PGI2 production by endothelial cells in vitro. It was shown that PGI2 production by cultured bovine aortic endothelial cells was significantly reduced in the presence of a high concentration of glucose (300 mg/dl) compared with physiological concentrations of glucose (100 mg/ dl). In contrast, no reduction in PGI2 production was observed in cells cultured with equimolar mannitol, suggesting that glucose itself, rather than the effect of osmolality, inhibited PGI2 production by cultured endothelial cells.In addition, a high concentration of glucose also inhibited the proliferation of cultured endothelial cells.

Angiotensin II increases the release of endothelin-I from human cultured endothelial cells but does not regulate its circulating levels

1999 ◽  
Vol 96 (3) ◽  
pp. 261-270 ◽  
Author(s):  
Claudio FERRI ◽  
Giovambattista DESIDERI ◽  
Roberta BALDONCINI ◽  
Cesare BELLINI ◽  
Marco VALENTI ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Protein Synthesis ◽  
Angiotensin Ii ◽  
At1 Receptor ◽  
Dependent Manner ◽  
Synthesis Inhibitor ◽  
Endothelin 1 ◽  
Cultured Endothelial Cells

We investigated the effect of angiotensin II on endothelin-1 secretion in vitro and in vivo. In vivo, angiotensin II was given intravenously to 23 essential hypertensive and 8 control subjects according to different protocols: Study A, 1.0 ngċmin-1ċkg-1 and 3.0 ngċmin-1ċkg-1 angiotensin II for 30 min each; Study B, 1.0 ngċmin-1ċkg-1 and 3.0 ngċmin-1ċkg-1 angiotensin II for 120 min each; Study C, 3.0 ngċmin-1ċkg-1 angiotensin II for 30 min followed by a dose increment of 3.0 ngċmin-1ċkg-1 every 30 min until mean blood pressure levels increased by 25 mmHg; Study D, 1.0 ngċmin-1ċkg-1 followed by 3.0 ngċmin-1ċkg-1 angiotensin II for 60 min each on two different NaCl diets (either 20 mmol NaCl/day or 220 mmol NaCl/day, both for 1 week). In all in vivo studies neither plasma nor urine endothelin-1 levels changed with angiotensin II infusion. In contrast, angiotensin II (10-9, 10-8, 10-7 mol/l) stimulated endothelin-1 secretion from cultured human vascular endothelial cells derived from umbilical cord veins in a time- and dose-dependent manner. The in vitro angiotensin II effects were abolished by candesartan cilexetil, an inhibitor of the membrane-bound AT1 receptor, and also by actinomycin D, an RNA synthesis inhibitor, and cycloheximide, a protein synthesis inhibitor, indicating that endothelin-1 release depended on AT1 receptor subtype and de novo protein synthesis. Our findings indicate that angiotensin II regulates endothelin-1 release by cultured endothelial cells through an AT1 receptor-dependent pathway, but does not influence circulating endothelin-1 levels in vivo.

Regulated release and functional modulation of junctional adhesion molecule A by disintegrin metalloproteinases

Blood ◽  
2009 ◽  
Vol 113 (19) ◽  
pp. 4799-4809 ◽  
Author(s):  
Rory R. Koenen ◽  
Jessica Pruessmeyer ◽  
Oliver Soehnlein ◽  
Line Fraemohs ◽  
Alma Zernecke ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Adhesion Molecule ◽  
Vascular Inflammation ◽  
Neutrophil Infiltration ◽  
Transendothelial Migration ◽  
Cultured Endothelial Cells ◽  
Leukocyte Transendothelial Migration ◽  
Functional Modulation ◽  
Air Pouch Model

Abstract Junctional adhesion molecule A (JAM-A) is a transmembrane adhesive glycoprotein that participates in the organization of endothelial tight junctions and contributes to leukocyte transendothelial migration. We demonstrate here that cultured endothelial cells not only express a cellular 43-kDa variant of JAM-A but also release considerable amounts of a 33-kDa soluble JAM-A variant. This release is enhanced by treatment with proinflammatory cytokines and is associated with the down-regulation of surface JAM-A. Inhibition experiments, loss/gain-of-function experiments, and cleavage experiments with recombinant proteases indicated that cleavage of JAM-A is mediated predominantly by the disintegrin and metalloproteinase (ADAM) 17 and, to a lesser extent, by ADAM10. Cytokine treatment of mice increased JAM-A serum level and in excised murine aortas increased ADAM10/17 activity correlated with enhanced JAM-A release. Functionally, soluble JAM-A blocked migration of cultured endothelial cells, reduced transendothelial migration of isolated neutrophils in vitro, and decreased neutrophil infiltration in a murine air pouch model by LFA-1– and JAM-A–dependent mechanisms. Therefore, shedding of JAM-A by inflamed vascular endothelium via ADAM17 and ADAM10 may not only generate a biomarker for vascular inflammation but could also be instrumental in controlling JAM-A functions in the molecular zipper guiding transendothelial diapedesis of leukocytes.

Plasma endothelin-1 response to acute hypotension induced by vasodilating agents

1994 ◽  
Vol 72 (9) ◽  
pp. 985-991 ◽  
Author(s):  
Peter Cernacek ◽  
Duncan J. Stewart ◽  
Mortimer Levy
Keyword(s):  
Endothelial Cells ◽  
Atrial Natriuretic Peptide ◽  
Natriuretic Peptide ◽  
Cyclic Gmp ◽  
Endothelin 1 ◽  
Arterial Blood ◽  
Cultured Endothelial Cells ◽  
Acute Hypotension ◽  
Atrial Natriuretic

Cyclic GMP (cGMP) dependent vasodilating agents (natriuretic peptides, nitric oxide) inhibit secretion of endothelin-1 (ET-1) in cultured endothelial cells. However, in circulatory conditions associated with acute hypotension, a marked increase in plasma ET-1 has repeatedly been observed. Therefore, after administration of cGMP-dependent agents in hypotensive dose, the net effect of these opposing influences on ET-1 release may shed light on the mechanisms determining circulating levels of this peptide. We have studied the effect of a hypotensive dose of atrial natriuretic peptide (n = 16), 8-Br-cGMP (n = 5), and papaverin (n = 7) on plasma ET-1 in anesthetized dogs. All agents produced marked increases in the peptide level at the end of infusion (178, 280, and 240% of the last preinfusion level, respectively) and a mean arterial blood pressure (MAP) decrease of 19, 18, and 42 mmHg (1 mmHg = 133.3 Pa), respectively. In all three protocols, plasma ET-1 continued to rise when the hypotensive agent was discontinued and remained elevated for 2–3 h postinfusion, even though MAP was normalized. There was a close positive correlation between the maximal increment in plasma ET-1 and the maximal decrease in MAP (r = 0.67, p < 0.001). These results show that acute hypotension due to directly acting vasodilators is a potent stimulus for systemic release of ET-1, even when due to agents known to inhibit ET-1 production in cultured endothelial cells. The discrepancy between the previous in vitro data and the present in vivo data can be explained either by non-endothelial origin of circulating ET-1, at least during acute hypotension, or by substantial differences in the production of ET-1 in the intact organism as opposed to endothelial cells in culture.Key words: endothelin secretion, big endothelin, atrial natriuretic peptide, cyclic GMP, papaverin.

scholarly journals Peptides derived from two separate domains of the matrix protein thrombospondin-1 have anti-angiogenic activity

1993 ◽  
Vol 122 (2) ◽  
pp. 497-511 ◽  
Author(s):  
SS Tolsma ◽  
OV Volpert ◽  
DJ Good ◽  
WA Frazier ◽  
PJ Polverini ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Granulation Tissue ◽  
Matrix Protein ◽  
Angiogenic Activity ◽  
Vessel Formation ◽  
Cultured Endothelial Cells ◽  
Thrombospondin 1

Thrombospondin-1 (TSP1) is a large modular matrix protein containing three identical disulfide-linked 180-kD chains that inhibits neovascularization in vivo (Good et al., 1990). To determine which of the structural motifs present in the 180-kD TSP1 polypeptide mediate the anti-angiogenic activity, a series of protease-generated fragments were tested using several in vitro and in vivo assays that reflect angiogenic activity. The majority of the anti-angiogenic activity of TSP1 resides in the central 70-kD stalk region which alone could block neovascularization induced by bFGF in the rat cornea in vivo and inhibit both migration in a modified Boyden chamber and [3H]thymidine incorporation stimulated by bFGF in cultured capillary endothelial cells. Although TSP1 has been shown to bind active TGF beta 1, this cytokine could not account for the inhibitory effects of the stalk region of TSP1 on cultured endothelial cells. Peptides and truncated molecules were used to further localize inhibitory activity to two domains of the central stalk, the procollagen homology region and the properdin-like type 1 repeats. Trimeric recombinant TSP1 containing NH2-terminal sequences truncated after the procollagen-like module inhibited endothelial cell migration in vitro and corneal neovascularization in vivo whereas trimeric molecules truncated before this domain were inactive as was the NH2-terminal heparin-binding domain that is present in both recombinant molecules. A series of peptides from the procollagen-like region, the smallest of which consisted of residues 303-309 of TSP1, inhibited angiogenesis in vivo in the rat cornea and the migration of endothelial cells in vitro. A 19-residue peptide containing these sequences blocked vessel formation in the granulation tissue invading a polyvinyl sponge implanted into the mouse. Nineteen residue peptides derived from two of the three type 1 repeats present in the intact TSP1 molecule blocked neovascularization in vivo in the rat cornea and inhibited the migration of cultured endothelial cells with ED50's of 0.6-7 microM. One of these peptides, containing residues 481-499 of TSP1, also inhibited vessel formation in granulation tissue invading sponges in vivo. These results suggest that the large TSP1 molecule employs at least two different structural domains and perhaps two different mechanisms to accomplish a single physiological function, the inhibition of neovascularization. The definition of short peptides from each of these domains that are able to block the angiogenic process may be of use in designing targeted inhibitors of the pathological neovascularization that underlies many diseases.

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