Mechanism of Stimulation of Glucose Transport by H2O2: Role of Phospholipase C

• Published in: Archives of biochemistry and biophysics 1999-02, Vol.362 (1), p.113-122
• Main Authors: Prasad, Rajesh K., Ismail-Beigi, Faramarz
• Format: Article
• Language: English
• Subjects: 3-O-Methylglucose - metabolism; 3T3 Cells; Animals; big mitogen-activated protein kinase (BMK1); Biological Transport - drug effects; Calcium-Calmodulin-Dependent Protein Kinases - antagonists & inhibitors; Calcium-Calmodulin-Dependent Protein Kinases - metabolism; Cell Line; Enzyme Activation - drug effects; genistein; Glucose - metabolism; Glut 1; herbimycin A; hydrogen peroxide; Hydrogen Peroxide - pharmacology; MAP Kinase Kinase 4; Mice; Mitogen-Activated Protein Kinase 7; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; phorbol-12-myristate-13-acetate (TPA); phospholipase C (PLC); Phosphotyrosine - metabolism; protein kinase C (PKC); Protein Kinase C - metabolism; Protein Kinases - metabolism; Protein-Tyrosine Kinases - antagonists & inhibitors; Rats; staurosporine; stress-activated protein kinase (SAPK); thapsigargin; Type C Phospholipases - metabolism; Type C Phospholipases - physiology; tyrosine kinase; U73122
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1006/abbi.1998.1026

Exposure of Clone 9 cells, a rat liver cell line, to hydrogen peroxide (H2O2) resulted in a striking and rapid stimulation of glucose transport (8- to 10-fold in 1 h). A comparable response was found in 3T3-L1 preadipocytes, C2C12myoblasts, and NIH 3T3 fibroblasts, which, similar to Clone 9 cells, express only the Glut 1 glucose transporter isoform. The enhancement of glucose transport in Clone 9 cells in response to H2O2was significantly attenuated by genistein and the phospholipase C (PLC) inhibitor, U73122. Exposure to H2O2resulted in a rise in cellsn-1,2-diacylglycerol content, and the rise was significantly inhibited by U73122. Moreover, the H2O2-induced stimulation of glucose transport was significantly blocked by thapsigargin. Neither staurosporine nor a 24-h preincubation in the presence of phorbol-12-myristate-13-acetate (TPA) affected the stimulatory effect of hydrogen peroxide on glucose transport. The activity of big mitogen-activated kinase (BMK1) and of stress-activated protein kinase (SAPK), both members of mitogen-activated protein kinases, were enhanced in response to exposure to H2O2; however, neither protein kinase appeared to be linked to the enhancement of glucose transport by H2O2. It is concluded that the stimulation of glucose transport in response to H2O2is independent of changes in PKC, BMK1, and SAPK activity, and is mediated, at least in part, through H2O2-induced stimulation of protein tyrosine kinase and PLC pathways.