Platelet Adhesion to Subendothelial Structures under Flow Conditions: No Effect of the Lipoxygenase Product 13-HODE

1989 ◽  
Vol 62 (02) ◽  
pp. 802-806 ◽  
Author(s):  
Jacques C de Graaf ◽  
Hidde Bult ◽  
Guido R Y de Meyer ◽  
Jan J Sixma ◽  
Philip G de Groot
Keyword(s):  
Endothelial Cells ◽  
Hydroxy Fatty Acid ◽  
Vessel Wall ◽  
Platelet Adhesion ◽  
Direct Evidence ◽  
Inhibitory Effect ◽  
Flow Conditions ◽  
Physiological Concentration ◽  
Hydroxyoctadecadienoic Acid ◽  
Wall Interaction

SummaryIt has been shown that endothelial cells can convert linoleic acid to 13-hydroxyoctadecadienoic acid (13-HODE) and it has been suggested that 13-HODE has non-thrombogenic properties. However, no direct evidence has been presented that indicates that 13-HODE indeed modulates platelet-vessel wall interaction.In this study we have bound a purified 13-HODE to a thrombogenic surface and its effect on platelet adhesion was studied and compared to the effects of an analogous hydroxy fatty acid, 15-hydroxyeicosatetraenoic acid (15-HETE). The effect of 13-HODE on platelet adhesion was studied both under static and flow conditions. In this report we show that binding of up to 40 times the physiological concentration to a thrombogenic surface has no inhibitory effect on platelet adhesion under static or flow conditions. We conclude that 13-HODE is not an important regulatory substance in platelet-subendothelium interaction, although this does not exclude it has a putative anti-adhesive role on intact endothelium.

Effect of Sulphinpyrazone (SP) Aspirin (ASA) and Dipyridamole (DP) on Plateletvessel Wall Interaction after Oral Administration to Rabbits

1979 ◽  
Author(s):  
J.A. Davies ◽  
V.C. Menys
Keyword(s):  
Clinical Trials ◽  
Platelet Aggregation ◽  
Vessel Wall ◽  
Platelet Adhesion ◽  
Ex Vivo ◽  
Inhibitory Effect ◽  
Inhibitory Effects ◽  
Human Dose ◽  
Wall Interaction ◽  
Adhesion Of Platelets

Clinical trials of anti-platelet drugs have suggested that they may be useful in the prevention of thrombotic disease. While such drugs inhibit platelet function, those which act on cyclooxygenase also reduce PGI2 synthesis and may interfere with tne natural antithrombotic properties of the vessel wall. We studied the effects of SP, ASA and OP ex vivo on the platelet-vessel wall interaction. Rabbits were dosed by mouth with drug (at about twice the weight-adjusted human dose) or placebo for 5 days, then exsanguinated and aortas removed. Washed platelets prepared from the blood were labelled with 51Cr. and their adhesion to everted aortapr epared from treated or control rabbits was measured in a perfusion device. PGI2-like activity in aortic rings was assayed by its inhibitory effect on platelet aggregation to ADP. Adhesion of platelets to aort as from SP- treated rabbits was i ncreased (p < 0.025), PGI2 - like activity was partially inhibited, but over all adhesion of SP-treated platelets to aor tas f rom SP-treated animals reduced by 30% (p < 0.02). Adhesion to aortas of ASA- treated rabbits was sliahtly inc r ea=-.ed (p > 0 . 1) , PGI 2 - l ike act ivi ty abolished , and no overall reduc tion in platelet adhesion seen. DP had no effecton adhesion or PGI-like activity. These results support the evidence that cyclo-oxygenase inhibitors reduce the inherent resistance of the vessel wall to platelet adhesion. However with SP, inhibitory effects on platelets appear to be more important.

scholarly journals Platelet-dependent primary hemostasis promotes selectin- and integrin- mediated neutrophil adhesion to damaged endothelium under flow conditions

Blood ◽  
1996 ◽  
Vol 87 (8) ◽  
pp. 3271-3281 ◽  
Author(s):  
PH Kuijper ◽  
HI Gallardo Torres ◽  
JA van der Linden ◽  
JW Lammers ◽  
JJ Sixma ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Vessel Wall ◽  
Platelet Adhesion ◽  
Leukocyte Adhesion ◽  
Umbilical Vein ◽  
Blood Platelets ◽  
Neutrophil Adhesion ◽  
Shear Stresses ◽  
Flow Conditions

Co-localization of blood platelets and granulocytes at sites of hemostasis and inflammation has triggered an intense interest in possible interactions between these cellular processes and induction of vessel wall injury. Leukocyte adhesion to endothelial cells decreases with increasing shear and is dependent on an initial rolling phase mediated by selectins. We hypothesized that flow-dependent platelet adhesion at an injured vessel wall will lead to P-selectin expression by platelets, thus mediating leukocyte co-localization. A perfusion chamber was used in which flowing whole blood induced platelet adhesion to a subendothelial matrix (ECM) of cultured human umbilical vein endothelial cells (HUVEC). We compared neutrophil (polymorphonuclear leukocyte [PMN]) interactions with HUVEC and their ECM with and without adhered platelets. PMNs adhered predominantly to ECM-adhered platelets and not to endothelial cells. ECM alone did not support PMN adhesion under flow conditions. PMN adhesion to unstimulated HUVEC was only substantial at low shear (up to 200 cells/mm2 at shear stress 80 mPa). In marked contrast, PMN adhesion to ECM-adhered platelets was dramatically increased, and adhesion was demonstrated at much higher shear stress (up to 640 mPa). Studies with specific antibodies showed that the platelet-dependent neutrophil adhesion was selectin-mediated. Inhibition of P-selectin caused a marked inhibition of adhesion at high shear stress, whereas the role of leukocyte L-selectin was less pronounced. beta2-Integrin-blocking antibodies inhibited static neutrophil adhesion. fMLP induced L-selectin shedding from leukocytes, resulting in decreased leukocyte adhesion. In conclusion, platelet- dependent hemostasis at the ECM appears to be a powerful intermediate in neutrophil-vessel wall interactions at shear stresses that normally do not allow neutrophil adhesion to intact endothelium.

Thrombin Inhibition and Thrombin Generation by Cultured Aortic Endothelial and Smooth Muscle Cells from Minipig

1982 ◽  
Vol 48 (01) ◽  
pp. 101-103 ◽  
Author(s):  
B Kirchhof ◽  
J Grünwald
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vessel Wall ◽  
Thrombin Generation ◽  
Muscle Cells ◽  
Thrombin Inhibition ◽  
Generating Capacity ◽  
Wall Interaction ◽  
Cells Cultured

SummaryEndothelial and smooth muscle cells cultured from minipig aorta were examined for their inhibitory activity on thrombin and for their thrombin generating capacity.Endothelial cells showed both a thrombin inhibition and an activation of prothrombin in the presence of Ca++, which was enhanced in the presence of phospholipids. Smooth muscle cells showed an activation of prothrombin but at a lower rate. Both coagulation and amidolytic micro-assays were suitable for studying the thrombin-vessel wall interaction.

IS THE ENDOTHELIAL EXTRACELLULAR MATRIX THROMBOGENIC OR THROMBORESISTANT? EFFECT OF PREPARATION AND 13-HODE LEVELS

1987 ◽  
Author(s):  
M R Buchanan ◽  
E Bastida ◽  
J Aznar-Salatti ◽  
P de Groot
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cell ◽  
Cellulose Acetate ◽  
Surface Area ◽  
Whole Blood ◽  
Hplc Analysis ◽  
Platelet Adhesion ◽  
Flow Conditions ◽  
Hydroxyoctadecadienoic Acid ◽  
Static Conditions

It is generally thought that the extracellular matrix (ECM) is thrombogenic.However,one of us (MRB) has reported that the ECM is thromboresistant,and postulated that this was due to the release of endothelial cell (EC) 13-hydroxyoctadecadienoic acid (13-HODE) into the ECM. To test this possibility, we measured platelet adhesion (PLT ADH) onto cultured ECs and their ECMs exposed by 3 methods. We also extracted the ECMs for HPLC analysis of 13-HODE.PLT ADH was expressed as i)adhesion of 3H-adenine labelled platelets/mm2 of ECs or ECMs under static conditions, and ii) % surface^ area coverage measured morphometrically following 5"perfusion with citrated whole blood at 1300 sec-1 in the flat chamber.ECMs were prepared by removing the EC monolayers by freeze thawing , cellulose acetate stripping or NH4OH treatment. PLT ADH to ECs under static and flow conditions were 4700±240/mm2 and 0.1%, respectively, and were associated with 12,6± 1 pg of 13-HODE/mm2 of EC surface (M+SEM). Removal of the ECs by freeze thawing or stripping, resulted in a 18% and 25% increase in PLT ADH to the ECM,under static and flow conditions respectively, and a 80% decrease in ECM associated 13-HODE level. Removal of the EC by NH4OH resulted in a 380% and 770% increase in PLT ADH to the ECM in static and flow conditions. 13-HODE was undetectable.These data support the hypothesis that 13-HODE released from ECs influences the ECM thrombogenecity, and indicate that the residual amounts of components present in the ECMs following EC removal is influenced by the method of ECM preparation.

Effects Of Aspirin And Sulfinpyrazone On Platelets, Vascular Cells And Their Interactions

1981 ◽  
Author(s):  
M R Buchanan ◽  
M J Vazquez ◽  
M A Gimbrone
Keyword(s):  
Growth Rate ◽  
Cell Growth ◽  
Dna Synthesis ◽  
Vessel Wall ◽  
Culture Medium ◽  
Platelet Adhesion ◽  
Thymidine Incorporation ◽  
Inhibitory Effect ◽  
Human Platelets ◽  
Final Density

Sulfinpyrazone (SUL) and aspirin (ASA) are potentially useful antithrombotic drugs. Both drugs are thought to exert this effect by inhibiting the platelet enzyme, cyclooxygenase (C-0), thus preventing thromboxane A2 synthesis. Recent data, however, suggest that these drugs also may affect vessel wall cells. To study this further, we examined the effects of SUL and ASA on i) the adhesion of 3H-adenine-labelled washed human platelets to cultured bovine endothelial (EC) and smooth muscle cells (SMC), ii) EC and SMC DNA synthesis (3H-thymidine incorporation) and iii) cell growth. Pretreatment of platelets with 100μM ASA or 250μM SUL (concentrations sufficient to inhibit C-0), did not affect platelet adhesion to untreated EC or SMC. However, adhesion of untreated, ASA- and SUL-platelets was increased 25,28 and 44% resp. when EC were pretreated with 650μM SUL for 24 hr. In contrast, adhesion of ASA-platelets to EC pretreated with lOOμM ASA (sufficient to inhibit prostacyclin), was unaffected. Platelet adhesion to SMC pretreated with 650μM SUL for 24 hr was decreased when platelets also were pretreated with ASA (20%, p<0.05) or SUL (27%, pc 0.02). Pretreatment of SMC with SUL for only 2 hr had no effect. DNA synthesis in EC and SMC treated with 62.5 and 250μM SUL for 24 hr, was inhibited >35% and >95% resp. Preliminary data suggest that this inhibitory effect may last longer in SMC. To study the effect of SUL on cell growth, EC and SMC were plated at 2 × 104 cells/ cm2 and fed with culture medium containing 0, 62.5 or 625uM SUL on day 0, 1, 3 and 4.5. EC growth rate and final density were unaffected over 7 days. SMC growth rate also was unaffected, but the final density of SMC treated with 650μM SUL was 31 μ 2% less than untreated SMC at 7 days (p<0.01). These data indicate that SUL has direct effects on EC and SMC that may influence i) platelet-vessel wall interactions and ii) vascular cell proliferation.

RELATIONSHIP BETWEEN VESSEL WALL 13-HODE PRODUCTION AND SUBENDOTHELIAL BASEMENT MEMBRANE THROMBORESISTANCE: INFLUENCE OF SALICYLATE

1987 ◽  
Author(s):  
E Weber ◽  
T A Haas ◽  
J Hirsh ◽  
M R Buchanan
Keyword(s):  
Endothelial Cells ◽  
Linoleic Acid ◽  
Basement Membrane ◽  
Vessel Wall ◽  
Carotid Arteries ◽  
Platelet Adhesion ◽  
Fold Increase ◽  
Lipoxygenase Pathway ◽  
The Relationship ◽  
Camp Levels

In previous studies, we have reported that i) the basement membrane (BM) underlying endothelial cells was initially throm-boresistant; ii) 13-hydroxyoctadecadienoic acid (13-HODE) synthesized by endothelial cells from linoleic acid via the lipoxygenase pathway, contributed to the thromboresistance of the endothelium; and iii) salicylate (SAL) increased injured vessel wall thrombogenecity. Therefore, we performed studies to determine the relationship between injured vessel wall thrombogenecity, vessel wall 13-HODE and cAMP levels, and salicylate treatment. Injured vessel wall thrombogenecity was measured as the number of H-adenine platelet (3H-PLT) adherent to the subendothelial BM exposed by air injury in carotid arteries of rabbits treated with 0 or 100 mg/kg of SAL bid, given orally2. Vessel wall 13-HODE was measured as the amount of 13-HODE/cm produced by the vessel wall following stimulation with 10μ/M linoleic acid, and measured by HPLC. Vessel wall cAMP levels were measured by RIA. Four hours after air injury, there was 25.4 ± 2 3 2H-PLT/cm2 of exposed BM. This was associated with 15.9 ng/cm2 of 13-HODE and 308 pM/cm2 of cAMP (Table 1). In contrast, in rabbits treated with SAL, there was a 2-fold increase in platelet adhesion onto the injured carotid arteries. The increase in platelet adhesion was associated with a 65% decrease in 13-HODE production by the vessel wall and a modest (20%) decrease in cAMP level.We conclude that the lipoxygenase derived linoleic acid metabolite, 13-HODE contributes not only to the thromboresis-tance of the endothelium, but also to its underlying basement membrane.

The contributions of the α2β1 integrin to vascular thrombosis in vivo

Blood ◽  
2003 ◽  
Vol 102 (10) ◽  
pp. 3652-3657 ◽  
Author(s):  
Li He ◽  
Loretta K. Pappan ◽  
David G. Grenache ◽  
Zhengzhi Li ◽  
Douglas M. Tollefsen ◽  
...  
Keyword(s):  
Vascular Injury ◽  
Vessel Wall ◽  
Platelet Adhesion ◽  
Thrombus Formation ◽  
Critical Role ◽  
Flow Conditions ◽  
In Vivo Models ◽  
Vascular Thrombosis ◽  
Α2β1 Integrin

AbstractThe α2β1 integrin serves as a receptor for collagens, laminin, and several other nonmatrix ligands. Many studies have suggested that the α2β1 integrin is a critical mediator of platelet adhesion to collagen within the vessel wall after vascular injury and that the interactions of the platelet α2β1 integrin with subendothelial collagen after vascular injury are required for proper hemostasis. We have used the α2β1 integrin-deficient mouse to evaluate the contributions of the α2β1 integrin in 2 in vivo models of thrombosis. Studies using a model of endothelial injury to the carotid artery reveal that the α2β1 integrin plays a critical role in vascular thrombosis at the blood-vessel wall interface under flow conditions. In contrast, the α2β1 integrin is not required for the formation of thrombi and pulmonary emboli following intravascular injection of collagen. Our results are the first to document a critical in vivo role for the α2β1 integrin in thrombus formation at the vessel wall under conditions of shear following vascular injury. (Blood. 2003;102:3652-3657)

scholarly journals Localization of 13-hydroxyoctadecadienoic acid and the vitronectin receptor in human endothelial cells and endothelial cell/platelet interactions in vitro

Blood ◽  
1993 ◽  
Vol 81 (12) ◽  
pp. 3303-3312 ◽  
Author(s):  
MR Buchanan ◽  
MC Bertomeu ◽  
TA Haas ◽  
FW Orr ◽  
LL Eltringham-Smith
Keyword(s):  
Endothelial Cells ◽  
Platelet Adhesion ◽  
Receptor Expression ◽  
Immunofluorescent Staining ◽  
Apical Surface ◽  
Binding Assays ◽  
Vitronectin Receptor ◽  
Wall Cell ◽  
Hydroxyoctadecadienoic Acid

Abstract Blood/vessel wall cell interactions depend, in part, on the expression of adhesion receptors on cell surfaces, such as expression of the vitronectin receptor (VnR) on the apical surface of endothelial cells (ECs) for platelet/EC adhesion. However, it is unclear how receptor expression is regulated from within cells. In previous studies, we found that ECs metabolize linoleic acid into the lipoxygenase monohydroxide, 13-hydroxyoctadecadienoic acid (13-HODE), and that the intracellular level of 13-HODE correlates inversely with VnR expression and platelet adhesion to the EC apical surface. In this study, we determined the physical associations of 13-HODE and VnR in unstimulated and stimulated ECs, ie, at times when ECs were and were not adhesive for specific ligands and platelets, using double antibody immunofluorescent staining techniques and binding assays. 13-HODE and the VnR were colocalized within unstimulated ECs. When ECs were stimulated, 13-HODE was no longer detectable, either in or outside the ECs, and the VnR was detected on the apical surface of the ECs. These changes were paralleled by increased vitronectin binding and increased platelet adhesion to the ECs. We suggest that colocalization of 13-HODE with VnR reflects a 13-HODE/VnR interaction, confining the VnR in a nonadhesive form inside unstimulated ECs, and, as a result, the ECs are nonadhesive. When the ECs are stimulated, 13-HODE and VnR dissociate, allowing the VnR to relocate on the EC surface, where the VnR undergoes a conformational change resulting in increased EC adhesivity.

Fibrillar cellular fibronectin supports efficient platelet aggregation and procoagulant activity

2015 ◽  
Vol 114 (12) ◽  
pp. 1175-1188 ◽  
Author(s):  
Eric Maurer ◽  
Mathieu Schaff ◽  
Nicolas Receveur ◽  
Catherine Bourdon ◽  
Luc Mercier ◽  
...  
Keyword(s):  
Vessel Wall ◽  
Platelet Adhesion ◽  
Thrombus Formation ◽  
Procoagulant Activity ◽  
Toll Like Receptor ◽  
Flow Conditions ◽  
Toll Like Receptor 4 ◽  
Cellular Fibronectin ◽  
Fibrillar Network ◽  
Platelet Functions

SummaryThe ability of cellular fibronectin, found in the vessel wall in a fibrillar conformation, to regulate platelet functions and trigger thrombus formation remains largely unknown. In this study, we evaluated how parietal cellular fibronectin can modulate platelet responses under flow conditions. A fibrillar network was formed by mechanically stretching immobilised dimeric cellular fibronectin. Perfusion of anticoagulated whole blood over this surface resulted in efficient platelet adhesion and thrombus growth. The initial steps of platelet adhesion and activation, as evidenced by filopodia extension and an increase in intracellular calcium levels (419 ± 29 nmol/l), were dependent on integrins α5β1 and αIIbβ3. Subsequent thrombus growth was mediated by these integrins together with the GPIb-V-IX complex, GPVI and Toll-like receptor 4. The involvement of Toll-like receptor 4 could be conveyed via its binding to the EDA region of cellular fibronectin. Upon thrombus formation, the platelets became procoagulant and generated fibrin as revealed by video-microscopy. This work provides evidence that fibrillar cellular fibronectin is a strong thrombogenic surface which supports efficient platelet adhesion, activation, aggregation and procoagulant activity through the interplay of a series of receptors including integrins α5β1 and αIIbβ3, the GPIb-V-IX complex, GPVI and Toll-like receptor 4.

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