Article

Mechanism underlying FeCl(3)-induced arterial thrombosis

UMR-S949 Inserm-Université de Strasbourg, Strasbourg, France.
Journal of Thrombosis and Haemostasis (Impact Factor: 5.55). 04/2011; 9(4):779-89. DOI: 10.1111/j.1538-7836.2011.04218.x
Source: PubMed

ABSTRACT The FeCl(3)-induced vascular injury model is widely used to study thrombogenesis in vivo, but the processes leading to vascular injury and thrombosis are poorly defined.
The aim of our study was to better characterize the mechanisms of FeCl(3)-induced vascular injury and thrombus formation, in order to evaluate the pathophysiological relevance of this model.
FeCl(3) was applied at different concentrations (from 7.5% to 20%) and for different time periods (up to 5 min) to mouse carotid or mesenteric arteries.
Under all the conditions tested, ultrastructural analysis revealed that FeCl(3) diffused through the vessel wall, resulting in endothelial cell denudation without exposure of the inner layers. Hence, only the basement membrane components were exposed to circulating blood cells and might have contributed to thrombus formation. Shortly after FeCl(3) application, numerous ferric ion-filled spherical bodies appeared on the endothelial cells. Interestingly, platelets could adhere to these spheres and form aggregates. Immunogold labeling revealed important amounts of tissue factor at their surface, suggesting that these spheres may play a role in thrombin generation. In vitro experiments indicated that FeCl(3) altered the ability of adhesive proteins, including collagen, fibrinogen and von Willebrand factor, to support platelet adhesion. Finally, real-time intravital microscopy showed no protection against thrombosis in GPVI-immunodepleted and β(1)(-/-) mice, suggesting that GPVI and β(1) integrins, known to be involved in initial platelet adhesion and activation, do not play a critical role in FeCl(3)-induced thrombus formation.
This model should be used cautiously, in particular to study the earliest stage of thrombus formation.

1 Follower
 · 
172 Views
 · 
2 Downloads
  • Source
    • "FeCl3 triggers an oxidative vascular endothelial matrix. Then, platelets interact with collagen and vWF in the matrix via their respective platelet surface receptors, leading to platelet adhesion [30]. According to the in vitro results, the in vivo antithrombotic efficacy of DJS was anticipated. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Dangguijagyagsan (DJS), a traditional herbal prescription, has long been used to treat menopause-related symptoms. We identified the cardioprotective effects of an aqueous extract of DJS using an ovariectomized (OVX) and ferric chloride- (FeCl-) induced carotid thrombosis rat model. Female Sprague-Dawley (SD) rats were ovariectomized or Sham-operated (Sham-control). The ovariectomized rats were divided into three groups: OVX with saline (OVX-control), aspirin 30 mg/kg/day (OVX-ASA), and DJS 100 mg/kg/day (OVX-DJS). The treatments were administered for 5 weeks. Then, blood samples were collected to analyze the serum lipid levels and platelet aggregation. The topical application of 40% FeCl3 induced intravascular thrombosis, which was used to test thrombotic occlusion and for histological examination. Body weight and the levels of total cholesterol (TC), triglyceride (TG), and LDL-cholesterol (LDL-C) increased in the OVX rats. These effects were reduced by ASA and DJS treatment. In addition, ASA and DJS treatment significantly inhibited platelet aggregation. These treatments also increased time to occlusion and decreased both thrombus size and the presence of collagen fibers in surrounding vessel walls compared with the Sham-control and OVX-control groups. These results suggest that DJS has beneficial effects in terms of preventing cardiovascular disease in menopausal woman because it can reduce the serum lipid levels and improve blood flow by inhibiting platelet aggregation and thrombus formation.
    Evidence-based Complementary and Alternative Medicine 09/2014; 2014:497836. DOI:10.1155/2014/497836 · 1.88 Impact Factor
  • Source
    • "Our methods had distinct features compared to other experimental models evaluating thrombolytic effects. First, to produce arterial thrombosis, we used a smaller filter paper saturated with a higher concentration (50%) of FeCl3 than previous studies (2.5–65%) [23,24,38,39]. This was because we needed to monitor blood flow for a longer period to examine the thrombolytic effects of drugs. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Saxatilin, a novel disintegrin purified and cloned from the venom of the Korean snake Gloydius saxatilis, strongly inhibits activation and aggregation of platelets. Glycoprotein (GP) IIb/IIIa receptor antagonists can resolve thrombus, so saxatilin might also have thrombolytic effects. We investigated the thrombolytic effects of saxatilin in mice using a ferric chloride-induced carotid arterial thrombosis model. Thrombotic occlusion and thrombus resolution were evaluated quantitatively by measuring blood flow in the carotid artery with an ultrasonic flow meter and calculating the degree of flow restoration on a minute-by-minute basis; results were confirmed by histological examination. Saxatilin dissolved thrombi in a dose-dependent manner. Saxatilin at 5 mg/kg restored blood flow to baseline levels. As saxatilin dose increased, time to recanalization decreased. A bolus injection of 10% of a complete dose with continuous infusion of the remaining dose for 60 minutes resulted in effective recanalization without reocclusion. The thrombolytic effect of saxatilin was also demonstrated in vitro using platelet aggregometry by administering saxatilin in preformed thrombi. Bleeding complications were observed in 2 of 71 mice that received saxatilin. Fibrin/fibrinogen zymography and platelet aggregometry studies indicated that saxatilin does not have fibrinolytic activity, but exerted its action on platelets. Integrin-binding assays showed that saxatilin inhibited multiple integrins, specifically α2bβ3 (GP IIb/IIIa), α5β1, αvβ3, αvβ1, and αvβ5, which act on platelet adhesion/aggregation. Saxatilin inhibited multiple integrins by acting on platelets, and was safe and effective in resolving thrombi in mice.
    PLoS ONE 11/2013; 8(11):e81165. DOI:10.1371/journal.pone.0081165 · 3.23 Impact Factor
  • Source
    • "In this model FeCl 3 is applied to the topical surface of an intact vessel, triggering vascular wall injury and denudation of the endothelium via a mechanism involving the generation of reactive oxygen species [4] [14]. A recent study suggested that FeCl 3 -induced vascular injury was erythrocyte-dependent , requiring hemolysis and hemoglobin oxidation for endothelial denudation [20] [21]. One measurable parameter in this model is the elapsed time from injury to complete vessel occlusion, measured as blood flow cessation by Doppler flowmeter or under direct observation with intravital microscopy [13] [14] [15]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Ferric chloride (FeCl3) induced vascular injury is a widely used model of occlusive thrombosis that reports platelet activation in the context of an aseptic closed vascular system. This model is based on redox-induced endothelial cell injury, which is simple and sensitive to both anticoagulant and anti-platelets drugs. The time required for platelet aggregation to occlude blood flow gives a quantitative measure of vascular damage that is pathologically relevant to thrombotic disease. We have refined the traditional FeCl3-induced carotid artery model making the data highly reproducible with lower variation. This paper will describe our artifices and report the role of varying the oxidative damage by varying FeCl3 concentrations and exposure. To explore a maximum difference between experimental groups, adjustment of the selected FeCl3 dose and exposure duration may be necessary.
    01/2013; 1(1):50-5. DOI:10.1016/j.redox.2012.11.001
Show more

Preview (2 Sources)

Download
2 Downloads