dexamethasone on lung mechanics and histology, inflammation, and apoptosis in the lung and distal organs in CLP-induced sepsis. The possible mechanisms

of action of both agents were also investigated, PCI-32765 focusing on oxidative stress (nuclear factor E2-related factor 2, GPx, CAT, iNOS, and SOD expression in lung tissue) and levels of interleukin (IL)-6, KC and IL-10 in bronchoalveolar lavage fluid (BALF). This study was approved by the local Animal Care Committee and conducted in compliance with the Guide for the Care and Use of Laboratory Animals (National Academy of Sciences, Washington, DC). Seventy-eight male BALB/c mice (20–25 g) were kept under specific pathogen-free conditions and a 12-h light/dark cycle in the Laboratory of Pulmonary Investigation animal care facility. All animals were randomly assigned to two groups. In the

control group (C), mice were subjected to sham surgery, while in the CLP group, cecal ligation and puncture was performed. Briefly, animals were anesthetized with ketamine (65 mg/kg, intraperitoneally [i.p.]) and xylazine (30 mg/kg, i.p.) and a midline laparotomy (2-cm incision) was performed. The cecum was carefully isolated to prevent damage to blood vessels. A 3-0 cotton ligature was placed below the ileocecal valve to prevent bowel obstruction. Finally, the cecum was punctured once with an 18-gauge needle and the animals left to recover from anesthesia (Oliveira et al., 2009 and Chao et al., 2010). In sham surgery, the abdominal cavity was opened and the cecum was isolated without ligation and puncture. The animals received subcutaneous injections of 1 mL of warm (37 °C) saline and buy Cyclopamine tramadol hydrochloride (20 mg/kg, i.p.). Both groups were further randomized to receive saline solution (SAL, 0.1 mL, i.p.), oleanolic acid (OA, 10 mg/kg, i.p.), or dexamethasone (DEXA, 1 mg/kg, i.p.) 1 h after sham or CLP surgery. Thirty-six mice (n = 6 per group) were selected for assessment of lung mechanics and histology; cell apoptosis in lung, kidney, aminophylline liver, and intestine samples; and measurement of CAT, GPx, iNOS, Nrf2 and SOD mRNA expression. The remaining

42 animals (n = 7/group) were subjected to the same protocol described above to obtain BALF aliquots for analysis. 24 h after sham or CLP surgery, animals were sedated (diazepam, 1 mg/kg, i.p.), anesthetized (thiopental sodium, 20 mg/kg, i.p.), tracheotomized, paralyzed (vecuronium bromide, 0.005 mg/kg, intravenously), and ventilated with a constant flow ventilator (Samay VR15; Universidad de la Republica, Montevideo, Uruguay) using the following settings: respiratory frequency 100 breaths min−1, tidal volume (VT) 0.2 mL, and fraction of inspired oxygen (FiO2) 0.21. A positive end-expiratory pressure (PEEP) of 2 cm H2O was applied and the anterior chest wall was surgically removed. After a 10-min ventilation period, static lung elastance (Est,L) was measured by the end-inflation occlusion method (Bates et al., 1985).