Vascular lipoxygenase activity: synthesis of 15-hydroxyeicosatetraenoic acid from arachidonic acid by blood vessels and cultured vascular endothelial cells

1987 ◽  
Vol 45 (6) ◽  
pp. 803-816 ◽  
Author(s):  
Hiroshi Takayama ◽  
Michael A. Gimbrone ◽  
Andrew I. Schafer
Keyword(s):  
Endothelial Cells ◽  
Arachidonic Acid ◽  
Blood Vessels ◽  
Vascular Endothelial Cells ◽  
Lipoxygenase Activity ◽  
Vascular Endothelial ◽  
Hydroxyeicosatetraenoic Acid

Cytochrome P450 Isoform Expression in Human Vascular Endothelial Cells

Hypertension ◽  
2000 ◽  
Vol 36 (suppl_1) ◽  
pp. 698-698
Author(s):  
Eduardo Barbosa-Sicard ◽  
Eva Kaergel ◽  
Dominik N Muller ◽  
Horst Honeck ◽  
Friedrich C Luft ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Arachidonic Acid ◽  
Cytochrome P450 ◽  
Vascular Endothelial Cells ◽  
Arachidonic Acid Metabolism ◽  
Vascular Endothelial ◽  
Rt Pcr ◽  
Pcr Screening ◽  
Mediators Of Inflammation ◽  
Derivatives Of

P29 Epoxy derivatives of arachidonic acid may act as important autocrine and paracrine mediators of endothelial function including regulation of vascular tone and control of inflammation. To identify potential candidates for catalyzing the synthesis of these and further arachidonic acid metabolites, we studied human vascular endothelial cells for the expression of individual cytochrome P450 isoforms belonging to the CYP families 1, 2, 3 and 4. An RT-PCR screening performed with subfamily- and isoform-specific primer pairs revealed mRNAs for the P450 forms 1A1, 1B1, 2C8, 2E1, 2J2, 3A7, 4A11 and 4F2. The identity of the RT-PCR products was confirmed by DNA sequencing. In addition, P450 1A2 mRNA was detected after induction with β-naphthoflavone which also enhanced the expresion of P450s 1A1 and 1B1. P450s 2B6 and 3A4 were not detectable. Similar P450 isoform patterns were obtained analyzing primary human endothelial cells originating from aorta, coronary arteries, dermal microvessels and umbilical veins, as well as an immortalized human endothelial cell line (HMEC-1). Further studies with HMEC-1 cells showed the expression of all human members of the P450 2C subfamily (2C8, 2C9, 2C18 and 2C19). We next used gaschromatography-mass spectrometry to identify the regioisomeric epoxeicosatrienoic acids produced by HMEC-1 cells. Among the P450 forms detected by the RT-PCR screening, P450 2C8 and 2J2 are the leading candidates for producing vasoactive epoxyeicosatrienoic acids. Using recombinant human P450 1A1, we then found that this P450 form catalyzes the formation of various regioisomeric hydroxy derivatives of arachidonic acid. We conclude that P450 1A1 known primarily for its role in polycyclic aromatic hydrocarbon metabolism, may interfere with endothelial arachidonic acid metabolism, particularly after its induction by drugs and xenobiotics. Furthermore, P450s 4A11 and 4F2 probably contribute to the degradation of lipid mediators of inflammation.

scholarly journals Arachidonic Acid Diols Produced by Cytochrome P-450 Monooxygenases Are Incorporated into Phospholipids of Vascular Endothelial Cells

1996 ◽  
Vol 271 (24) ◽  
pp. 14001-14009 ◽  
Author(s):  
Mike VanRollins ◽  
Terry L. Kaduce ◽  
Xiang Fang ◽  
Howard R. Knapp ◽  
Arthur A. Spector
Keyword(s):  
Endothelial Cells ◽  
Arachidonic Acid ◽  
Vascular Endothelial Cells ◽  
Vascular Endothelial ◽  
Cytochrome P 450

scholarly journals TGF-β1 Potentiates the Cytotoxicity of Cadmium by Induction of a Metal Transporter, ZIP8, Mediated by the ALK5-Smad2/3 and ALK5-Smad3-p38 MAPK Signal Pathways in Cultured Vascular Endothelial Cells

2021 ◽  
Vol 23 (1) ◽  
pp. 448
Author(s):  
Keisuke Ito ◽  
Tomoya Fujie ◽  
Masahiro Shimomura ◽  
Tsuyoshi Nakano ◽  
Chika Yamamoto ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Blood Vessels ◽  
Transforming Growth Factor ◽  
Vascular Endothelial Cells ◽  
Extracellular Fluid ◽  
Mitogen Activated Protein Kinase ◽  
Signal Pathways ◽  
Vascular Endothelial ◽  
Metal Transporter ◽  
Tgf Β1

Vascular endothelial cells cover the luminal surface of blood vessels in a monolayer and play a role in the regulation of vascular functions, such as the blood coagulation-fibrinolytic system. When the monolayer is severely or repeatedly injured, platelets aggregate at the damaged site and release transforming growth factor (TGF)-β1 in large quantities from their α-granules. Cadmium is a heavy metal that is toxic to various organs, including the kidneys, bones, liver, and blood vessels. Our previous study showed that the expression level of Zrt/Irt-related protein 8 (ZIP8), a metal transporter that transports cadmium from the extracellular fluid into the cytosol, is a crucial factor in determining the sensitivity of vascular endothelial cells to cadmium cytotoxicity. In the present study, TGF-β1 was discovered to potentiate cadmium-induced cytotoxicity by increasing the intracellular accumulation of cadmium in cells. Additionally, TGF-β1 induced the expression of ZIP8 via the activin receptor-like kinase 5-Smad2/3 signaling pathways; Smad3-mediated induction of ZIP8 was associated with or without p38 mitogen-activated protein kinase (MAPK). These results suggest that the cytotoxicity of cadmium to vascular endothelial cells increases when damaged endothelial monolayers that are highly exposed to TGF-β1 are repaired.

Prostacyclin: Production by Vascular Endothelium and Effects on Platelet Aggregation

1979 ◽  
Author(s):  
S. Moncada ◽  
S. Bunting
Keyword(s):  
Endothelial Cells ◽  
Arachidonic Acid ◽  
Platelet Aggregation ◽  
Vascular Endothelium ◽  
Vascular Endothelial Cells ◽  
Specific Inhibitor ◽  
Inhibitory Effect ◽  
Vascular Endothelial ◽  
Prostacyclin Production

The inhibitory effect of vascular endothelial cells on platelet aggregation is due to their ability to release prostacyclin. The existence of an ADPase has been confirmed in endothelial cells but this enzymes does not seem to be related to the anti-aggregating properties of vascular endothelium. In vitro, the release of prostacyclin by humand and rabbit endothelial cells persists after several subcultures. The production of PGI2 can be demonstrated by its inhibition by aspirin-like drugs or 15-hydroperoxy arachidonic acid (a specific inhibitor of PGI2 synthesis). Moreover, the antiaggregating activity is antagonised by an antibody to 5,6 dihydro prostacyclin which cross reacts and neutralises prostacyclin.

Extracellular ATP signaling and P2X nucleotide receptors in monolayers of primary human vascular endothelial cells

2002 ◽  
Vol 282 (2) ◽  
pp. C289-C301 ◽  
Author(s):  
Lisa M. Schwiebert ◽  
William C. Rice ◽  
Brian A. Kudlow ◽  
Amanda L. Taylor ◽  
Erik M. Schwiebert
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Blood Vessels ◽  
Vascular Tone ◽  
Vascular Endothelial Cells ◽  
Purinergic Signaling ◽  
Primary Cultures ◽  
Atp Release ◽  
Vascular Endothelial ◽  
Multiple Blood

ATP and its metabolites regulate vascular tone; however, the sources of the ATP released in vascular beds are ill defined. As such, we tested the hypothesis that all limbs of an extracellular purinergic signaling system are present in vascular endothelial cells: ATP release, ATP receptors, and ATP receptor-triggered signal transduction. Primary cultures of human endothelial cells derived from multiple blood vessels were grown as monolayers and studied using a bioluminescence detection assay for ATP released into the medium. ATP is released constitutively and exclusively across the apical membrane under basal conditions. Hypotonic challenge or the calcium agonists ionomycin and thapsigargin stimulate ATP release in a reversible and regulated manner. To assess expression of P2X purinergic receptor channel subtypes (P2XRs), we performed degenerate RT-PCR, sequencing of the degenerate P2XR product, and immunoblotting with P2XR subtype-specific antibodies. Results revealed that P2X4and P2X5are expressed abundantly by endothelial cell primary cultures derived from multiple blood vessels. Together, these results suggest that components of an autocrine purinergic signaling loop exist in the endothelial cell microvasculature that may allow for “self-regulation” of endothelial cell function and modulation of vascular tone.

scholarly journals Exogenous H2S Regulates of Cystathionine Gamma-Lyase in HUVECs During Hypoxia

2020 ◽  
Author(s):  
Maoxian Wang
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Mrna Expression ◽  
Blood Vessels ◽  
Western Blotting ◽  
Promoter Activity ◽  
Vascular Endothelial Cells ◽  
Vascular Endothelial Cell ◽  
Luciferase Assay ◽  
Vascular Endothelial

Cystathionine gamma-lyase (CSE) is one of the essential H2S-producing enzymes, and it regulates diverse functions in connection with cardiovascular function. It is crucial how exogenous H2S regulates CSE expression of the vascular endothelial cell during hypoxia. We examined the transcription and expression of CSE in HUVECs regulated by exogenous H2S with 100 μM during hypoxia by Luciferase assay, Western blotting, and quantitative RT-qPCR. Exogenous H2S influenced on the promoter activity of CSE in HUVECs during hypoxia. The effects of 100 μM H2S on CSE mRNA expression in HUVECs is decreased compared with 0 μM H2S. The consequences of 100 μM H2S on the expression level of CSE protein in HUVECs at two h of hypoxia is reduced compared with 0 μM H2S. These findings suggest that vascular endothelial cells can respond to the signals of hypoxia in the blood, and can respond to changes in H2S concentration in the blood, thus affect the blood vessels themselves.

Measurement Of 1-14C Arachidonic Acid Metabolism By Cultured Human Vascular Endothelial Cells And Platelets

1981 ◽  
Author(s):  
Ch Willems ◽  
P J Laanen ◽  
G A Pool ◽  
Ph G de Groot ◽  
J A van Mourik ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Arachidonic Acid ◽  
Acid Metabolism ◽  
Vascular Endothelial Cells ◽  
Lipid Fraction ◽  
Relative Sensitivity ◽  
Arachidonic Acid Metabolism ◽  
Vascular Endothelial ◽  
Response Curves ◽  
Stimulation Of

To investigate the relative sensitivity of endothelial cells and platelets towards aspirin (ASA), a method was designed which allows the measurement of prostaglandin (PG) metabolism using [l-14C] arachidonic acid (AA) as precursor. When HEC were incubated with 20 μM AA for 24h at 37°C in the presence of serum, about 15-20% of the label was incorporated into phospholipids whereas 3-4% was present in the neutral lipid fraction. No PG formation could be measured, even after stimulation of the phospholipase system by thrombin or ionophore A 23187. When HEC were incubated for 20 min with AA in the absence of serum, about 8-10% of the label was incorporated into PGs (6-keto PGF1α, 2%; PGE2, 1%; PGF2α, 4%). Platelets were prelabeled with AA, in buffer and the phospholipase system could be stimulated by thrombin (measured as the formation of TXB2, HHT and HETE). Using the above methods, the IC50 of ASA (30 min, 37°C) was 4.7±2.4μM for HEC and 10.1±2.4μM for platelets (p>0.1). Preincubation with 5μM ASA resulted in 50% inhibition of PG synthesis after 30±8 min with HEC and after 40±5.6 min with platelets (p#x003E;0.1). Moreoever when HEC and platelets were combined, incubated with ASA and subsequently tested separately no difference in dose-response curves could be demonstrated. These results indicate that the sensitivity of HEC and platelets to ASA is similar.

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