Hydrogen peroxide increases [ ³ H]-2-deoxyglucose uptake via MAPKs, cPLA ₂ , 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 [ ³ H]-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-κB. AACOCF ₃ or mepacrine (cPLA ₂ inhibitors, 10 ^-6 M), SN 50 (NF-κB nucleus translocation inhibitor, 500 ng/ml) or Bay11-7082 (a IκB-α phosphorylation inhibitor, 2×10 ^-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-κB signaling pathways in mouse ES cells.