INFLUENCE OF HYPOXIA AND HYPOXIA-REOXYGENATION ON ENDOTHELIAL P-SELECTIN EXPRESSION

doi:10.1016/S0049-3848(96)00233-2

Thrombosis Research

Volume 85, Issue 2, 15 January 1997, Pages 159-164

https://doi.org/10.1016/S0049-3848(96)00233-2 Get rights and content

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Cells culture.

Endothelial cells were isolated from human umbilical cord vein (HUVEC) by the method of Jaffe [13]. Cells were grown to confluence in Medium 199 (Gibco) supplemented with 10% of human serum (J. Boy), 10% fetal calf serum (FCS, Biological Industrie), 1% Hepes (Gibco), 1% Penistreptomycin (Gibco), 0.06% Fungizone (Squib). HUVECs were subcultured by brief exposure to 0.1% trypsin and 0.005% EDTA (Boerhinger) and grown to confluence in 35 mm diameter wells (6 or 24 wells Cluster, Costar) in 20% FCS

Effect of hypoxia on endothelial cell surface expression of P-selectin (Fig. 1)

Modulation of endothelial P-selectin expression has been evaluated as a function of hypoxic exposure time. HUVEC have been exposed to hypoxia for 30, 60, 90, 120 or 240 minutes. Our data indicated that hypoxia induced P-selectin expression on the endothelial surface (Fig. 1a). The level of P-selectin expression rapidly increased since the fluorescence intensity was 166±33% after 30 minutes of hypoxic exposure. It reached a maximum at 90 minutes (278±7%), and then moderatly declined, the

DISCUSSION

Because of their localization at the interface between blood and tissues, endothelial cells are the first cells to undergo damages when exposed either to hypoxia or to hypoxia followed by reoxygenation. They respond by striking phenotype modulation, becoming prothrombotic [15]and proinflammatory 16, 17, 18. One consequence of this phenotype modulation is an increase of PMN adherence to the endothelium. Studies using animal models have evidenced the key role of P-selectin in PMN adhesion induced

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Cited by (30)

Hypoxic stress -responsive genes in air breathing catfish, Clarias magur (Hamilton 1822) and their possible physiological adaptive function
2016, Fish and Shellfish Immunology
Citation Excerpt :
Among them, the complement component 3 (C3) is important for the development of pulmonary arterial hypertension in hypoxia exposed mice [48]. Hypoxia has also been found to initiate the coagulation cascade via the activation of P selectin and tissue factors [49]. Amara et al. 2010 [47] suggested the coagulation cascade and complement system are linked together, where the activation of coagulation cascade led the activation of other via cleavage of C3 and C5.
The Indian catfish, Clarias magur (previous name C. batrachus) is an air breathing fish, inhabitant of aquatic bodies characterized by low dissolved oxygen levels. It is exposed to hypoxic conditions in its natural habitat. Thus, it can be useful model to study the mechanism of hypoxia stress tolerance. In C. magur, molecular processes facilitating its adaptation to hypoxia stress remain largely unexplored, in part due to unavailability of genomic resources. The suppression subtractive hybridization technique (SSH) was employed to compare the differential expression of transcripts under experimental hypoxic conditions, to that of normoxic conditions. Twelve subtracted cDNA libraries (six each forward and reverse) were constructed from brain, heart, liver, muscle, spleen and head kidney tissues. A total of 2020 clones were screened and sequenced, resulting into 1805 high quality expressed sequence tags (ESTs). Annotation of these differentially expressed ESTs resulted into the identification of genes involved in vast majority of pathways/processes affecting metabolism, cellular processes, signal transduction and/or immune functions. Additionally, 18 potential novel genes expressed in hypoxia stress exposed fish were also identified. The study had catalogued the differentially expressed genes from hypoxia stress induced C. magur, where most of them are reported for the first time in a hypoxia-tolerant fish species. The results not only provided insights for the hypoxia stress altered cellular functions in C. magur, but also generated a valuable functional genomics resource to assist targeted studies on functional genomics and future genome projects.
Simulated thrombin responses in venous valves
2016, Journal of Vascular Surgery: Venous and Lymphatic Disorders
Venous thromboembolism frequently results in thrombi formation near or within the pocket of a venous valve due to recirculating hemodynamics, which has been largely attributed to hypoxia-induced tissue factor (TF) expression. Numerical models are now capable of assessing the spatiotemporal behavior of the TF-initiated coagulation cascade under nonuniform hemodynamics. The aim of this study was to use such a numerical simulation to analyze the degree and location of thrombin formation with respect to TF position in the presence of disturbed flow induced by an open venous valve.
Thrombin formation was simulated using a computational model that captures the hemodynamics, kinetics, and chemical transport of 22 biochemical species. Disturbed flow is described by the presence of a valve in the equilibrium phase of the valve cycle with leaflets in a fully open position. Three different positions of TF downstream of the valve opening were investigated.
The critical amount of TF required to initiate a thrombotic response is reduced by up to 80% when it is positioned underneath the recirculating regions near the valve opening. In addition, because of the increased surface area of the open valve cusp in conjunction with recirculating hemodynamics, it was observed that thrombin is generated inside the valve pocket even when the exposed region of TF is downstream of the valve.
The presence of prothrombotic surface reactions in conjunction with recirculating hemodynamics provides an additional mechanism for thrombus formation in venous valves that does not require direct damage or dysfunction to the valve itself.
Molecular mechanisms underlying synergistic adhesion of sickle red blood cells by hypoxia and low nitric oxide bioavailability
2014, Blood
Citation Excerpt :
In this regard, it is interesting to note that the adhesion scores of eNOS-deficient mice at normoxia (Figure 3A, lane 5) are comparable to those of control mice under hypoxia (Figure 3A, lane 3). Because P-selectin expression is upregulated by low oxygen tension31 as well as by low NO bioavailability32 in multiple types of cells, it is reasonable to expect such results. We also observed an increase in leukocyte rolling influx, which is mediated by P-selectin.33
The molecular mechanisms by which nitric oxide (NO) bioavailability modulates the clinical expression of sickle cell disease (SCD) remain elusive. We investigated the effect of hypoxia and NO bioavailability on sickle red blood cell (sRBC) adhesion using mice deficient for endothelial NO synthase (eNOS) because their NO metabolite levels are similar to those of SCD mice but without hypoxemia. Whereas sRBC adhesion to endothelial cells in eNOS-deficient mice was synergistically upregulated at the onset of hypoxia, leukocyte adhesion was unaffected. Restoring NO metabolite levels to physiological levels markedly reduced sRBC adhesion to levels seen under normoxia. These results indicate that sRBC adherence to endothelial cells increases in response to hypoxia prior to leukocyte adherence, and that low NO bioavailability synergistically upregulates sRBC adhesion under hypoxia. Although multiple adhesion molecules mediate sRBC adhesion, we found a central role for P-selectin in sRBC adhesion. Hypoxia and low NO bioavailability upregulated P-selectin expression in endothelial cells in an additive manner through p38 kinase pathways. These results demonstrate novel cellular and signaling mechanisms that regulate sRBC adhesion under hypoxia and low NO bioavailability. Importantly, these findings point us toward new molecular targets to inhibit cell adhesion in SCD.
Surgery and Hemostasis
2013, Consultative Hemostasis and Thrombosis: Third Edition
Basic mechanisms and pathogenesis of venous thrombosis
2009, Blood Reviews
Citation Excerpt :
Further studies are needed to explore the possibility that changes in the ratio of procoagulant to anticoagulant properties of the valves make a contribution to venous thrombotic risk. In addition to modulating the pro and anticoagulant properties of the endothelium, hypoxia also up regulates the expression of P-selectin on endothelium leading to the recruitment of leukocytes or leukocyte microparticles containing tissue factor which can serve as the nidus for initiation of the thrombotic response18,19 (Fig. 2). Microparticles bearing tissue factor appear to play a role in thrombus formation.20,21
In 1856 Virchow proposed a triad of causes for venous thrombosis, postulating that stasis, changes in the vessel wall or changes in the blood could lead to thrombosis. We now know that abnormally high levels of some coagulation factors and defects in the natural anticoagulants contribute to thrombotic risk. Among these, factor V Leiden, which renders factor Va resistant to activated protein C, is the most prevalent with approximately 5% of the Caucasian population having this genetic alteration. These genetically controlled variants in coagulation factors work in concert with other risk factors, such as oral contraceptive use, to dramatically increase thrombotic risk. While these abnormalities in the blood coagulation proteins are associated with thrombotic disease propensity, they are less frequent contributors to thrombosis than age or cancer. Cancer increases thrombotic risk by producing tissue factor to initiate coagulation, by shedding procoagulant lipid microparticles or by impairing blood flow. Age is the strongest risk factor for thrombosis. Among possible reasons are fragility of the vessels potentially contributing to stasis, increased coagulation factor levels, impaired function of the venous valves, decreases in the efficacy of natural anticoagulants associated with the vessel wall, increased risk of immobilization and increased risk of severe infection.
Pathophysiology of venous thrombosis
2009, Thrombosis Research
Using Virchow's triad as framework, it is clear that alterations in any of its components (blood composition, the vessel wall, and blood flow) can influence the propensity for the development of venous thromboembolism. Each can also influence the others in ways that enhance or reduce thrombotic propensity. While past work has concentrated on blood components that influence thrombogenicity, their influences generally are manifest only after the thrombotic process has begun, and it is initiation of thrombosis on the vessel wall that is the least well understood. This brief review attempts to link risk factors such as stasis and cancer to the mechanisms that initiate thrombosis.

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BRIEF COMMUNICATIONINFLUENCE OF HYPOXIA AND HYPOXIA-REOXYGENATION ON ENDOTHELIAL P-SELECTIN EXPRESSION

Section snippets

Cells culture.

Effect of hypoxia on endothelial cell surface expression of P-selectin (Fig. 1)

DISCUSSION

First page preview

J. Biol. Chem.

Cell

Cell

Trends Cardiovasc Med

Control of vascular permeability by polymorphonuclear leukocytes in inflammation

Nature Lond.

Effect of hypoxia on adherence of granulocytes to endothelial cells in vitro

Am. J. Physiol.

PADGEM (GMP-140) is a component of Weibel-Palade bodies of human endothelial cells

Blood

Increased PMN adherence on endothelial cells after hypoxia : involvement of PAF, CD18/CD11b, and ICAM-1

Am. J. Physiol.

Coexpression of GMP-140 and PAF by endothelium stimulated by histamine or thrombin : a justacrine system for adhesion and activation of neutrophils

J. Cell Biol.

PMN adherence to cat ischemic-reperfused mesenteric vascular endothelium under flow : role of P-selectin

J. Appl. Physiol.

Role of P-selectin in leukocyte-endothelial interaction in splanchnic ischemia-reperfusion

Am. J. Physiol.

BRIEF COMMUNICATION
INFLUENCE OF HYPOXIA AND HYPOXIA-REOXYGENATION ON ENDOTHELIAL P-SELECTIN EXPRESSION