001 N NaOH (Jiang et al , 2007) Two groups (n = 7 and n = 15) we

001 N NaOH (Jiang et al., 2007). Two groups (n = 7 and n = 15) were used for oxidative stress and behavioral experiments, respectively. They were randomly divided into

three groups: (1) Sham + vehicle group; (2) CLP + vehicle and (3) CLP + GUA group. We do not have a sham group with GUA in order to decrease the number of animals used, but we did some pilot experiments that in the conditions that GUA was administered in sham animals cognitive and oxidative damage parameters were not altered and in this study, we did not evaluate the GUA role in the sham group. ABT-199 in vivo Animals were submitted only to one behavioral task. Twelve or 24 h after the surgery procedure (CLP or sham), all rats were killed by decapitation. The hippocampus, cerebellum, striatum, prefrontal cortex and “cortex” (cerebral cortex without the prefrontal cortex) were quickly isolated by hand dissection Dasatinib molecular weight using a magnifying glass and a thin brush; dissection was based on histological distinctions described by Paxinos and Watson (1986). Samples were stored at −80 °C for subsequent analysis of oxidative stress. As an index of oxidative stress effects on lipids, we used the formation of TBARS during an acid-heating reaction, as previously described (Esterbauer and Cheeseman, 1990). Briefly, the samples were mixed with 1 mL

of 10% trichloroacetic acid and 1 mL of 0.67% thiobarbituric acid, and then heated in a boiling water bath for 15 min. TBARS was determined by the absorbance at 535 nm using 1,1,3,3-tetramethoxypropane HSP90 as an external standard. Results are expressed as malondialdehyde equivalents per milligram of protein. The oxidative stress effect on proteins was

assessed by the determination of carbonyl groups based on the reaction with dinitrophenylhidrazine, as previously described (Levine et al., 1994). Briefly, proteins were precipitated by the addition of 20% trichloroacetic acid and dissolved in dinitrophenylhidrazine, and the absorbance was read at 370 nm. Results are expressed as protein carbonylation per milligram of protein. Proteins were measured by the Lowry method (Lowry et al., 1951). The animals were separately submitted to four behavioral tasks: habituation to an open-field apparatus, inhibitory avoidance task, object recognition task and forced swimming task, 10 days after surgery. All behavioral procedures were conducted between 13:00 and 16:00 h in a sound-isolated room, and a single animal performed only one behavioral task in only one time point after surgery. All behavioral tasks were recorded by the same person who was blind to the animal group. This task evaluates aversive memory. The apparatus and procedures have been described in previous reports (Barichello et al., 2005 and Tuon et al., 2008). Briefly, the training apparatus was a 50 × 25 × 25 cm acrylic box (Insight, Brazil) whose floor consisted of parallel caliber stainless steel bars (1 mm diameter) spaced 1 cm apart. A 7 cm-wide, 2.

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