PGE2 and LTB4 are AA-derived metabolites from pathways dependent on cyclooxygenase (COX) and 5-lipoxygenase (5-LO), respectively (Peters-Golden and Brock, 2000; Samuelson, 2000; Funk, 2001). These lipid mediators are involved in inflammation and several homeostatic biological functions, including vascular permeability GSK2118436 concentration and leukocyte influx to the bronchoalveolar fluid (Teixeira et al., 1997; Nascimento et al., 2005). PGE2 is involved in the inflammatory response, and in the neutrophil recruitment (Fruscella et al., 2001) in mice inoculated with T. serrulatus scorpion venom ( Pessini et al., 2006). PGE2 is also produced after
i.p. inoculation of phospholipase A2 from the Bothrops asper snake venom in mice ( Moreira et al., 2011). Additionally, the action of crotoxin (neurotoxin isolated from Crotalus durissus terrificus venom) is modulated by 5-LO-derived lipidic mediators in rats ( Nogueira-Neto et al., 2008). However, there
is a lack of knowledge regarding the participation of these lipid mediators in cell recruitment to the peritoneal cavity induced by T. serrulatus Ts2 or Ts6. To address this question, we first demonstrated the kinetics of cell recruitment to the peritoneal cavity of mice injected with Ts2 or Ts6 isolated from the venom of scorpion T. serrulatus, and characterized the possible inflammatory mediators involved in cell migration. Second, we inhibited PGs and LTs synthesis by treatment with celecoxib, a COX-2 inhibitor, or MK-886, a 5-LO activation protein (FLAP) inhibitor, and characterized the cell types and cell recruitment MS-275 nmr kinetics
to the peritoneal cavity of mice injected with Ts2 or Ts6. Toxins Ts2 and Ts6, representing 3% and 2.5% of the total crude soluble TsV, respectively, were purified and stored at −20 °C as previously described (Arantes et al., 1989; Cologna et al., 2011, 2012). Prior to the Janus kinase (JAK) experiments, Ts2 and Ts6 were dissolved in phosphate buffered saline (PBS) and filtered through sterilizing membranes (Spritzenfilter: 0.22 mm, TPP, Switzerland). To determine whether the purified toxins were contaminated by the endotoxin LPS, a Limulus Amoebocyte Lysate test (LAL) was performed according to the manufacturer’s instructions (QCL-1000, Bio Whittaker, Cambrex Company, Walkersville, MD, USA). Male 129sv mice (6–8 weeks old) were obtained from the animal facility of Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP) – Universidade de São Paulo (USP). Male 5-LO deficient (5 LO−/−) mice were obtained from The Jackson Laboratory (Bar Harbor, ME, USA) and raised at FCFRP-USP with their age-matched male wild type littermates (WT-background, strain 129). These mice were maintained under standard laboratory conditions. All experiments were approved and conducted in accordance with the guidelines of the University Animal Care Committee (process n0 09.1.847.53.4). Groups of six mice were injected i.p. with 300 μL of Ts2 or Ts6 (250 μg/kg) diluted in sterile PBS.
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