CONSUMPTION OF BROCCOLI SPROUTS INCREASED THE ACTIVITY OF GLUTATHIONE-DEPENDENT ANTIOXIDANT ENZYMES IN MURINE LIVER

Abstract

Aim. The purpose of this work was to investigate effects of the consumption of broccoli sprouts on the activity of antioxidant defense enzymes in the liver of. GPx isoforms. After photo fixation of gels, we determined activity of each isoform densitometrically using ImageJ software. Results were statistically analyzed by ANOVA followed by post-hoc Tukey's test. Data are presented as mean±SEM. Methods. Frozen tissue was homozeniged in lysis buffer, centrifuged and resulted supernatants were used for analysis. Activities of enzymes were determined by using electrophoresis in polyacrylamide gel. After electrophoresis we conducted dyeing of separate gels for detection of superoxide dismutase (SOD), glutathione-S-transferase (GST) or glutathione peroxidase (GPx) isoforms. After photo fixation of gels, we determined activity of each isoform densitometrically using ImageJ software. Results were statistically analyzed by ANOVA followed by post-hoc Tukey's test. Data are presented as mean±SEM. Results. In the hepatic tissue of all four groups of mice, two isoforms of SOD (SOD1 and SOD2) were detected in gels. The intensity of the bands of both isoforms was not significantly different between groups. Three isoforms of GST (GST1, GST2, GST3) were detected in the liver samples. The activity of GST1 did not significantly differ between the experimental groups. Activities of GST2 and GST3 forms were significantly higher in the group “Broccoli” compared to Control and Cafeteria Diet groups. We identified three isoforms of glutathione peroxidase (GPx1, GPx2, GPx3) in liver samples. The activity of GPx isoform 1 was not significantly different between the experimental groups. The activity of GPx2 was significantly higher in the group of mice that consumed Broccoli and Cafeteria Diet + Broccoli compared to Control. GPx2 activity was significantly higher in the Broccoli group compared to the Cafeteria Diet group. Activity of GPx3 was significantly higher in the Broccoli group compared to the Control and Cafeteria Diet group. Conclusions. Cafeteria diet did not significantly affect the activity of SOD isoforms, but led to redistribution of in the activity of GST and GPx isoforms in murine liver. Feeding with broccoli spouts significantly increased the activity of 2 and 3 isoforms of GST and GPx in murine liver compared to values in control mice and mice fed with cafeteria diet. Combination of Broccoli + Cafereria diet had small activating effects on antioxidant enzyme acivity, compared with cafeteria diet.