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

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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.