A Method of Measuring Glutathione Peroxidase Activity in Murine Brain in Pharmacological Experiments

Resumo

A method of measuring of glutathione peroxidase activity using H₂O₂ was adapted for homogenates of murine brains. If the amount of reduced glutathione was at the constant level of 0.55 mM, the concentration of H₂O₂ of 0.192 mM was saturating for glutathione peroxidase of murine brain and was selected as an optimal concentration for the estimation of enzyme activity in tris-HCl buffer with addition of NaN₃ and EDTA (pH 8.5) at the incubation temperature of 37°C. The homogenates were dissolved by the reaction mixture by 10.4 times. The duration of incubation did not exceed 60 sec, if 13% homogenate was used. The experiment based on this method showed increased activity of glutathione peroxidase in the brain of mice treated with a derivative of acetaldehyde ammonia during long-term intermittent normobaric hypoxia. These data might reflect activation of glutathione peroxidase.