Haemostatic imbalance following carrageenan-induced rat paw oedema

Dipartimento Farmacologia Sperimentale, Università degli Studi di Napoli Federico II, via D. Montesano 49, 80 131 Napoli, Italy.
European Journal of Pharmacology (Impact Factor: 2.53). 01/2008; 577(1-3):156-61. DOI: 10.1016/j.ejphar.2007.08.007
Source: PubMed


Carrageenan-induced rat paw oedema is a widely used model to investigate the physiopathology of an acute local inflammation. Recently, much attention has been focused on the link between haemostasis and inflammation, and on the impact that inflammation might have on thrombotic events. It is known that the systemic response to inflammation is the "acute phase reaction" that represents a highly complex reaction of the organism to a variety of injuries, aimed to restore homeostasis; one important feature of the acute phase reaction is the hepatic synthesis of proteins involved in the coagulation cascade. Much attention has been focused on the role that systemic inflammation might have on thrombotic events, while there is not much information on the role played by an acute local inflammation on haemostasis, that can lead toward a pro-thrombotic state. The present study was conducted to evaluate the haemostatic balance in the early and the late phase of carrageenan-induced rat paw oedema; i.e. at 3 h, when paw inflammation is maximally expressed, and 24 h following carrageenan injection, when there is an almost complete absence of local inflammatory symptoms. We found that in inflamed animals, 24 h following oedema induction, there was an increase in plasma fibrinogen levels, antithrombin III activity and serum interleukin-6 levels, concomitant to a shortened prothrombin time and to an increased platelet responsiveness to ADP. Furthermore, in inflamed tissues at 3 h there was an increase in antithrombin III proteic expression. Our results demonstrate that a haemostatic imbalance occurs following carrageenan-induced rat paw oedema.

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Available from: Valentina Vellecco, Oct 08, 2015
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    • "Blood anticoagulated with 3.2% sodium citrate (1:9 citrate/blood, v/v) was withdrawn from male Wistar rats (anesthetized by enflurane) by cardiac puncture. Platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared as previously described (Pearce et al., 2004; Cicala et al., 2007). Briefly, PRP was obtained by centrifugation at 800 rpm for 15 min at 25 1C. "
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    ABSTRACT: Ethnopharmacological relevance: The roots of Salvia milthorriza Bunge (Lamiaceae) known as "Danshen", are used in Traditional Chinese Medicine as a remedy for activating blood and eliminating stasis. TIIA, a diterpenoid of Salvia milthorriza, is one of active components in Danshen that exhibits a significant improvement of the blood flow in the coronary circulatory system and a reduction of myocardial infarction. However, its effect on platelet and underlying mechanism remains largely unknown. On this basis, this compound could be a promising agent to improve blood viscosity and microcirculation and to prevent CVD. Materials and methods: In order to investigate the effects of TIIA on platelet functionality and its interaction with various platelet activation pathways, rat PRP were incubated with TIIA for 1 min at 37°C prior the addition of the stimuli (ADP or collagen). Aggregation was monitored in a light transmission aggregometer measuring changes in turbidity with continuous observation up to 10 min after the addition of the stimuli. MAPK signaling pathway and tubulin acetylation were analyzed by a Western blot technique. The effect of the TIIA was also studied in vivo on bleeding time in mice. Results: TIIA selectively inhibited rat platelet aggregation induced by reversible ADP stimuli (3 μM) in a concentration-dependent manner (0.5-50 μM). Nevertheless, TIIA was less active against the irreversible stimuli induced by ADP (10 μM) and collagen (10 μg/mL). Moreover, experiments performed on platelet lysates collected at different time-point after the addition of the stimuli shown that TIIA modulated tubulin acetylation and inhibited Erk-2 phosphorylation. Concomitantly, TIIA administrated i.p. at 10 mg/kg significantly amplified the mice bleeding time with an increase of 58% compared to its control (2.06±0.29 min vs 1.30±0.07). ASA was used as reference drug for in vitro and in vivo experiments. Conclusions: This study clarifies the intracellular signaling pathway involved in antiplatelet action of TIIA and also gives preliminary evidences for its anticoagulant activity. On this basis, this compound could be a promising agent to improve blood viscosity and microcirculation and to prevent CVD.
    Journal of Ethnopharmacology 07/2014; 155(2). DOI:10.1016/j.jep.2014.07.010 · 3.00 Impact Factor
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    • "Such outcomes are suggestive of a possibility that circulating IL-6 could act as a messenger of information from peripheral inflammatory sites to the CNS [23]. Furthermore, another study demonstrated that IL-6 serum levels were significantly increased at 24 hr following edema induction, but not after 3 hr, in a model of carrageenan-induced rat paw edema [24]. However, contradictory results indicated that LPS-induced inflammatory paw edema in rats, but not the type induced by carrageenan, resulted in measurable levels of IL-6 in serum within 3 hr of induction [25]. "
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    ABSTRACT: In the present study, we investigated the potential beneficial impact of the addition of antioxidant supplements to diclofenac regimen in a model of carrageenan-induced paw. Rats were treated daily with antioxidants, that is, a-lipoic acid (50 mg/kg), selenium (2.5 mg/kg), vitamin C (1 g/kg), vitamin E (300 mg/kg), or zinc (25 mg/kg) on seven successive days and then received a single treatment with diclofenac or saline before carrageenan was injected to induce paw inflammation. The results indicated that these combinations did not significantly affect the percentage inhibition of paw edema caused by diclofenac alone; however, some combination treatments ameliorated signs of concomitant oxidative stress (such as alterations in plasma malondialdehyde (MDA) levels, hemolysate reduced glutathione levels, and erythrocytic superoxide dismutase enzyme activities) imparted by diclofenac alone. In some cases, few tested antioxidants in combination with diclofenac resulted in increased plasma levels of interleukin- (IL-) 6 and C-reactive protein (CRP). In conclusion, the results of these studies suggested to us that the added presence of natural antioxidants could be beneficial as standard anti-inflammatory therapeutics for a patient under diclofenac treatment, albeit that these effects do not appear to significantly build upon those that could be obtained from this common anti-inflammatory agent per se.
    The Scientific World Journal 03/2014; 2014:731462. DOI:10.1155/2014/731462 · 1.73 Impact Factor
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    • "Carrageenan is a polysaccharide formed by repeated monomers that have groups of galactose sulfate, extracted from the cell wall of Rhodophyta algae such as Chondrus crispus (Campo et al., 2009). It has being widely used as an inductor of inflammation (Morris, 2003; Cicala et al., 2007, Vieira et al., 2012a,b). Inflammatory agents have been used to induce tendinopathies in experimental models used to study the efficacy of various potential therapies. "
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    ABSTRACT: Tendinopathy is a pathology found mainly in the rotator cuff, patellar, Achilles and flexor tendons. Tendinopathy is a significant impediment to performance in athletes and in workers in the labor market. Some studies have indicated that inflammation in adjacent tissues may affect the rotator cuff and Achilles tendon. In this study alterations were verified in the extracellular matrix (ECM) of the deep digital flexor tendon after two periods (12 and 24 hr) of induction inflammation in rat paw. Wistar rats were divided into three groups: those that received injection of 1% carrageenan; those that received 0.9% NaCl; and those that received no application. The tendon was divided into distal (d), proximal (p), and intermediate (i) regions. Biochemical analyses were performed and included non-collagenous proteins (NCP), glycosaminoglycans (GAGs), hydroxyproline (HoPro) and metalloproteinases 2 and 9. Tissue sections were stained with toluidine blue, hematoxylin-eosin, and Ponceau SS and observed under polarization microscopy. Remarkable results were detected that included the presence of MMP-9, degradation of NCP and GAG and the presence of cellular infiltrate closer to digits in d region. The different concentrations of HoPro, as well as alterations in the organization of the collagen fibers showed the collagenous matrix undergoing some alterations. The results indicated that the induced inflammation in rat paw exhibited characteristics similar to the typical acute inflammatory process observed in tendons. Anat Rec, 2013. © 2013 Wiley Periodicals, Inc.
    The Anatomical Record Advances in Integrative Anatomy and Evolutionary Biology 08/2013; 296(8). DOI:10.1002/ar.22729 · 1.54 Impact Factor
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