Impaired production of nitric oxide, superoxide, and hydrogen peroxide in glucose 6‐phosphate‐dehydrogenase‐deficient granulocytes

Abstract

Since the generation of superoxide and hydrogen peroxide by NADPH oxidase and nitric oxide (NO) by NO synthase (NOS) in granulocytes is NADPH-dependent, we investigated the production of NO, superoxide and H P O P in glucose 6-phosphate dehydrogenase (G6PD)-deficient human granulocytes. Our results showed that upon stimulation with either 5 W Wg/ml of lipopolysaccharide (LPS) or 10 W WM of phorbol 12-myristate 13-acetate (PMA), the production of nitrite in normal granulocytes was elevated, 252 þ 135% and 239 þ 72%, respectively, compared to the resting stage. In contrast, G6PD-deficient granulocytes did not produce more nitrite upon stimulation with either LPS or PMA compared to the resting stage. Western blot analysis indicated a normal expression pattern of inducible NOS in G6PD-deficient granulocytes. In addition, the production of H P O P and superoxide was also significantly impaired in G6PD-deficient granulocytes compared to control cells. These data demonstrate that G6PD deficiency causes an impairment in the production of NO, superoxide and H P O P .z 1998 Federation of European Biochemical Societies.

The Involvement of Glucose Metabolism in the Regulation of Inducible Nitric Oxide Synthase Gene Expression in Glial Cells: Possible Role of Glucose-6-Phosphate Dehydrogenase and CCAAT/Enhancing Binding Protein

“…The involvement of G6PD in iNOS gene expression was supported further by the inhibition of NO production and iNOS protein expression by the treatment with the antisense DNA oligomer of G6PD mRNA. Although these results indicated the involvement of PPP in glucose-mediated upregulation of LPS-induced expression of iNOS, several previous studies have suggested that the inhibition or deletion of G6PD decreases the LPS-induced NO production without altering the iNOS protein levels (Laychock and Bauer, 1996;Tsai et al, 1998). Because NADPH plays a role, as a cofactor, in the synthesis of NO, NO production is reduced by NADPH depletion evoked by PPP inhibition at an enzymatic level.…”

Macrophage Glucose-6-Phosphate Dehydrogenase Stimulates Proinflammatory Responses with Oxidative Stress

“…For example, it has been demonstrated that activation or overexpression of G6PD increases NO production in RINm5F rat insulinoma cells and bovine aortic endothelial cells (38,39). Also, it has been reported that inhibition of G6PD decreases ROS and RNS in various cell types, including macrophages (40), granulocytes (20), and bovine colonary arteries (41). Therefore, it is very likely that the physiological roles of G6PD might be dependent on cell types that have distinct microenvironments, with different levels of pro-oxidative and antioxidative enzymes.…”

“…Nitrite was measured using the Griess reaction (20). Culture media (100 l) were collected and incubated with an equal volume of Griess reagent for 10 min at room temperature.…”

Increase in Glucose-6-Phosphate Dehydrogenase in Adipocytes Stimulates Oxidative Stress and Inflammatory Signals

“…Measurement of cellular nitrite, hydrogen peroxide, and ROS levels. Nitrite was measured using the Griess reaction (25). Culture media (100 l each) were collected and incubated with an equal volume of Griess reagent for 10 min at room temperature.…”