Synergistic activation of glucose-6-phosphate dehydrogenase and NAD(P)H oxidase by Src kinase elevates superoxide in type 2 diabetic, Zucker fa/fa, rat liver

Glucose metabolism through the glycolysis and hexosamine pathway has been shown to be altered ill type 2 diabetes. However, the fate of glucose through the pentose phosphate pathway (PPP) is Currently unclear. In this study, we determined whether the activity of ghlcose-6-phosphate dehydrogenase (G6PD), the rate-limiting enzyme in the PPP, is modulated in the liver of zucker obese fa/fa rats (9-11 weeks of age). We found that G6PD expression and activity, NADPH levels, and 6-phosphogluconate generation were significantly increased in the liver of fa/fa rats. Inhibition of PI3 kinase and Src kinases decreased (p<0.05) G6PD activity in the fa/fa but not in the lean rat liver, Suggesting that G6PD activity is regulated by PI3/Si-c kinase signaling pathways. G6PD-derived NADPH increased (p<0.05) Superoxide anion levels by 70-90% in fa/fa vs lean rat liver, which was inhibited by the NADPH oxidase inhibitor gp91(ds-tat) (50 mu M) and G6PD inhibitors 6-aminonicotinamide (1 mM) and dehydroepiandrosterone ( 100 mu M), therefore indicating that elevated G6PD activity may be responsible for mediating superoxide generation. Interestingly, we also found a positive correlation between liver hypertrophy/increased G6PD activity (r(2) = 0.77; p = 0.0009) and liver hypertrophy/Superoxide production (r(2) = 0.51; p = 0.0091) in fa/fa rats. Increased G6PD and NADPH oxidase expression and activity, in young hyperglycemic and hyperinsulinemic rats before the development of diabetes, seems to be a contributing factor in the induction Of oxidative stress. Because inhibition of G6PD activity decreases oxidative stress, we conclude that G6PD behaves as a pro-oxidant in the fa/fa rat liver in type 2 diabetes. (C) 2009 Elsevier Inc. All rights reserved.