Oxidative Stress Evaluation in Pup Brain

The cerebral cortex and hippocampus of PND15 (n = 5 pups in each group) were individually dissected and mechanically homogenized in PBS. Afterward, the homogenates were centrifuged at 12,500 rpm for 30 min at 4°C with a Z216MK microcentrifuge (HERMLE Labortechnik GmbH; Wehingen, Germany). Aliquots of the supernatants were used to quantify nitrites, lipid peroxidation, GSH and GSSG levels, and enzymatic activities of AST, ALT, LDH, SOD, CAT, and GPx. Protein content was measured in the supernatants using the Coomassie blue method [32 (link)] to normalize the biochemical results.