Antioxidant Analysis in Mice Jejunum

According to Tsakiris et al. (40 (link)), pieces of mice jejuna from all experimental groups were weighed and homogenized to produce 10% (w/v) homogenate in an ice-cold solution containing 300 mM sucrose and 50 mM Tris(hydroxymethyl)aminomethane-HCl (Sigma, St Louis, MO, USA). The homogenate was centrifuged at 500×g at 4 °C for 10 min. The supernatant was separated and used in a variety of biochemical analyses. According to the technique used by Beutler et al. (41 ), the concentrations of reduced glutathione (GSH) were calculated using the absorbance measurement at 405 nm and expressed as mg/g. The level of glutathione peroxidase (GPx) was assessed using the Pagila and Valentine (42 ) method, which reported values as U/g. Lipid peroxide by-product as malondialdehyde (MDA) contents (43 (link)) were estimated in the supernatant of jejunum homogenate at 534 nm and represented as nmol/g. The Montgomery and Dymock (44 ) procedure were used to measure the level of nitric oxide (NO), which was then represented as µmol/L. The quantity of superoxide dismutase (SOD) was quantified at 560 nm and displayed as U/g (45 (link)). All these parameters were measured using the proper kits (Biodiagnostic Co., Egypt), which were supported by the software of SoftMax^® Pro v. 6.3.1. The molecular device used for the analysis was the Spectra MAX 190.