INTRODUCTION: Inflammation is a complex biological response that occurs in the body as a reaction to harmful stimuli [1]. The side effects caused by anti-inflammatory drugs have led scientists to search for therapeutical alternatives [2]. N-acylhydrazones have been widely used in medicinal chemistry and pharmacology due to their diverse biological properties, including antimicrobial [3], immunomodulatory and anti-inflammatory activities [4]. AIMS: The present work aimed to evaluate the anti-inflammatory activity of the N-acylhydrazone derivative 2-cyano-N’-(3-ethoxy-4-hydroxybenzylidene)-acetohydrazide (AMH) using a mice model of pleurisy induced by different phlogistic agents (carrageenan, zymosan, histamine and bradykinin), and to measure the levels of cytokines, nitric oxide (NO) and myeloperoxidase (MPO) in the pleural exudate. METHODS: N-acylhydrazone derivative (AMH) was provided by Prof. Dr. Ricardo Olímpio Moura from the Pharmacy Department at the State University of Paraíba (UEPB). This project was approved by the Ethics Committee on the Use of Animals of the Federal University of Pernambuco (CEUA-UFPE), under protocol number 23076.030374/2014-85. Swiss mice from the vivarium of the Department of Antibiotics at the Federal University of Pernambuco were used. Pleurisy was induced by injecting phlogistic, via intrapleural route, with one of the following agents: carrageenan (1% 0,1 ml/cav.) bradykinin (10 nmoles/cav.), histamine (100 µg/cav.) or zymosan (200 mg µg/cav.). The quantification of cytokines IL-1? and TNF-? in exudates was determined by the Sandwich ELISA protocol. To assess NO production, nitrate concentrations were measured using Griess reagent assay and later Sodium Nitrate Standard Curve [5]. The activity of the MPO enzyme was evaluated using the methodology described by Bradley et al. (1982) [6]. RESULTS AND DISCUSSION: AMH (10 mg/kg) decreased the concentration of TNF-? near the standard drug value, dexametasone (9,5 pg/mL), it also reduced IL-1? production compared to the non-treated control. IL1-? induces adhesion molecules, leukocyte migration, and increased vascular permeability, indicating it is an important pro-inflammatory mediator, besides that the release of these two pro-inflammatory cytokines may have been reduced due to the anti-inflammatory activity of AMH [7]. The results in carrageenan-induced pleurisy showed that AMH inhibited leukocyte migration in the thoracic cavity by 83,0% in comparison to the control group. At a dose of 10 mg/kg of AMH there was a significant reduction in the levels of NO (~67%) and MPO compared to the control group, suggesting that AMH acts by inhibiting the concentration of TNF-?, consequently reduces the production of NO. In the pleurisy test induced by histamine and zymosan, the AMH significantly inhibited leukocyte migration in 49,2% and 69,8%, respectively, in contrast with the control group. In bradykinin-induced pleurisy, the inhibition of leukocyte migration was 20,6% compared to the control group. CONCLUSION: AMH exhibited a promising anti-inflammatory activity by decreasing cell migration in pleurisy induced by carrageenan, zymosan, and histamine, however, there was no decrease in leukocyte migration in pleurisy induced by bradykinin. It also significantly decreased the levels of TNF-?, IL-1?, NO and MPO.
ACKNOWLEDGEMENT: Thanks to my laboratory colleagues, supervisor, and the Federal University of Pernambuco and CNPq for financial support.
REFERENCES
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Comissão Organizadora
Francisco Mendonça Junior
Pascal Marchand
Teresinha Gonçalves da Silva
Isabelle Orliac-Garnier
Gerd Bruno da Rocha
Comissão Científica
Ricardo Olimpio de Moura
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