Hydrogen Peroxide Increases [³H]-2-Deoxyglucose uptake via MAPKs, cPLA₂, and NF-κB Signaling Pathways in Mouse Embryonic Stem Cells

Abstract

Hydrogen peroxide (H₂O₂) has been shown to act as a signaling molecule that is involved in many cellular functions. This study investigated the effect of H₂O₂ on the [³H]-2-deoxyglucose (2-DG) uptake and its related signaling pathways in mouse embryonic stem (ES) cells. H₂O₂ significantly increased the level of 2-DG uptake in a time- (> 4 hr) and concentration- (>10^-4 M) dependent manner due to an increase in V_max but not K_m. Indeed, H₂O₂ increased the mRNA and protein level of glucose transporter 1 (GLUT 1). PD 98059 (a p44/42 MAPKs inhibitor, 10^-5 M), SB 203580 (a p38 MAPK inhibitor, 10^-6 M), and SP 600125 (a SAPK/JNK inhibitor, 10^-6 M) blocked the H₂O₂-induced increase in 2-DG uptake. H₂O₂ also increased phosphorylation of p44/42 mitogen activated protein kinases (MAPKs), p38 MAPK, and stress-activated protein kinase/Jun-N-terminal kinase (SAPK/JNK). In addition, H₂O₂ stimulated the translocation of cytosolic phospholipase A₂ (cPLA₂) from the cytosolic fraction to the membrane fraction, the release of arachidonic acid, and the activation of NF-κB. AACOCF₃ or mepacrine (cPLA₂ inhibitors, 10^-6 M), SN 50 (NF-κB nucleus translocation inhibitor, 500 ng/ml) or Bay11-7082 (a IκB-α phosphorylation inhibitor, 2x10^-5 M) blocked the H₂O₂-induced increase in 2-DG uptake. H₂O₂ increased the protein level of glucose transporter 1 (GLUT 1), which was blocked by PD 98059, SB 203580, SP 600125, mepacrine, or Bay11-7082. In conclusion, H₂O₂ increases the 2-DG uptake via MAPKs, cPLA₂, and NF-κB signaling pathways in mouse ES cells.

Beyond DNA binding - a review of the potential mechanisms mediating quinacrine's therapeutic activities in parasitic infections, inflammation, and cancers

“…Further support of this mechanism was attained by Na et al . when hydrogen peroxide-induced phosphorylation of the p65 subunit of NF-κB was partially inhibited by quinacrine [269]. The effects of quinacrine on NF-κB are in line with its uses in the treatment of inflammatory diseases as a single agent or in combination with other medications [21].…”

Upregulation of NF-κB upon differentiation of mouse embryonic stem cells

“…A potential contribution of NF-κB signaling to self renewal of human ES cells was suggested based on the higher expression level of phospho-p65(Ser536), as determined by immunostaining and confocal microscopy, along with the results obtained from inhibitor studies (32). NF-κB signaling was also suggested to be involved in the increase of glucose uptake upon hydrogen peroxide treatment in mouse ES cells, which was determined by immunostaining and inhibitor studies (33). Taken together with its low expression in ES cells, an attempt to increase NF-κB expression or activity in ES cells without impairing their stemness may be one of strategy for achieving better maintenance of ES cells.…”

Effect of dihydrotestosterone on hydrogen peroxide‐induced apoptosis of mouse embryonic stem cells

“…Among the ROSs, intracellular H 2 O 2 has been shown to induce detrimental effects on cells (Dumont et al, 1999;Liu et al, 2002;Park et al, 2005). Indeed, oxidative stress in tissues and cells has been generally recognized as a pivotal factor in the pathogenesis of embryonic development (Hausburg et al, 2005;Burton et al, 2006;Na et al, 2007). Therefore, the inhibition of such free radical-mediated pathophysiology has become a central focus of research efforts targeted toward the prevention or amelioration of embryo or ES cells injury.…”