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

Hydrogen peroxide (H(2)O(2)) has been shown to act as a signaling molecule that is involved in many cellular functions. This study investigated the effect of H(2)O(2) on the [3H]-2-deoxyglucose (2-DG) uptake and its related signaling pathways in mouse embryonic stem (ES) cells. H(2)O(2) 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, H(2)O(2) 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 H(2)O(2)-induced increase in 2-DG uptake. H(2)O(2) 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, H(2)O(2) stimulated the translocation of cytosolic phospholipase A(2) (cPLA(2)) from the cytosolic fraction to the membrane fraction, the release of arachidonic acid, and the activation of NF-kappaB. AACOCF(3) or mepacrine (cPLA(2) inhibitors, 10-6 M), SN 50 (NF-kappaB nucleus translocation inhibitor, 500 ng/ml) or Bay11-7082 (a IkappaB-alpha phosphorylation inhibitor, 2x10-5 M) blocked the H(2)O(2)-induced increase in 2-DG uptake. H(2)O(2) 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, H(2)O(2) increases the 2-DG uptake via MAPKs, cPLA(2), and NF-kappaB signaling pathways in mouse ES cells.