Table 2- Chemical composition (DM) of purslane used in the study
For determination of SOD and GPx activity and TAA and MDA concentration, erythrocytes
were and mixed with cold redistilled water and
stand at +4 °C for 15 minutes to lyses and then stored in 1.5-mL microtubes at −80°C. Hemoglobin (Hb) concentration was analyzed colorimetrically with a ZiestChem Diagnostic
kit (catalog no. 10-532, Ziest Chemistry, Tehran, Iran) for determination of SOD and GPx
activity (U/g Hb) and TAA and MDA concentration. Erythrocytes samples were analyzed for superoxide dismutase (SOD) activity with a SOD
Assay Kit (RANSOD, RANDOX, United Kingdom). The methodology used in the kit described
by Woolliams et al. (1983). This method employs xanthine and xanthine oxidase (XOD) to
generate superoxide radicals which react with 2-(4-iodophenyl)-3-(4-nitrophenol)-5- phenyltetrazolium chloride (INT) to form a red formazan dye, which measured by
Photometer. The superoxide dismutase activity is then measured by the degree of inhibition
reduction of INT under the conditions of the assay.
Glutathione peroxidase (GPx) activity was determined with a GPx Assay Kit (RANSEL, RANDOX, United Kingdom). The methodology used in the kit is described by Paglia and
Glutathione is immediately converted to the reduced form with a concomitant oxidation of
NADPH to NADP+
. The decrease in absorbance at 340 nm was measured at 37°C.
The concentration of malondialdehyde (MDA), an index of lipid peroxidation and oxidative
stress was determined in the erythrocyteas thiobarbituric acid reactive substances(TBARS)
by the method of Buege and Aust (1978).In the assay 200 μl of lysed erythrocyte sample was
mixed with 800 μl dH2O and 2ml of a TCA-TBA-HCl reagent (containing 15% trichloroacetic
acid (TCA), 0.375% thiobarbituric acid (TBA) and 0.25 N Hydrochloric acid (HCl) in distilled