This research was conducted to examine the effect of vernonia amygdalina on Glutathione reductase and Glutathione-S-transferase in alloxan induced diabetic wistar rats. Sixty-six (66) adult male wistar rats weighing between 100 and 250 g were used for the study. The animals were induced by intraperitoneal injection of 60 mg of alloxan. The animals were divided into 11 groups of six (6) rats in phase 1 and phase 2 (normal control, crude extract, Metformin, glycoside, saponin and alkaloid) respectively. After 28days treatment, the rats were sacrificed using a gaseous anesthetic agent (chloroform), blood samples were collected from inferior Vena Cava for biochemical analysis and pancreas harvested for histopathological examination. The final fasting blood glucose level of the experimental animals showed no significant difference when compared to that of the control rats (at p < 0.05). The level of Glutathione-S-transferase activities in the liver (2.93 ± 0.30) showed significant decrease in rats treated with various extracts and fractionates when compared with non-diabetic, untreated rats (control), whereas the GST activity in the brain (0.97 ± 0.10) and kidney (1.59 ± 0.80) of rats in all groups showed no significant difference when compared to their respective control group (p < 0.05). Activity of Glutathione reductase in the liver, serum, brain, and kidney show no significant difference in rats treated with various extract and fractionates when compared with nondiabetic, untreated rats (control). Vernonia amygdalina extract possess Hepatoprotective, Cytoprotective, hypoglycemic properties, protects and normalizes activities of Glutathione-S-transferase and Glutathione reductase.