Interleukin 1 stimulates hexose transport in fibroblasts by increasing the expression of glucose transporters
Bird, T.A.; Davies, A.; Baldwin, S.A.; Saklatvala, J.
Journal of Biological Chemistry 265(23): 13578-13583
ISSN/ISBN: 0021-9258 PMID: 2380175 Accession: 007476697
Exposure of quiescent cultures of human gingival fibroblasts (HuGi) and porcine synoviocytes (PSF) to human recombinant interleukin 1.alpha. or -.beta. (IL1.alpha. and -.beta.) enhnaced the rate of glycolysis as judged by increased lactate production. The cytokines also increased uptake of [3H]2-deoxyglucose (DG) in a time- and dose-dependent manner. Stimulation of DG uptake was first 6-8 h following addition of IL1 and was maximal by 24-30 h. IL1.alpha. and -.beta. were equipotent. Half-maximal stimulation occurred at approximately 1 pM IL1; maximal stimulation (2.5-4.5-fold in HuGi, 3-7-fold in PSF) was obtained with approximately 80 pM IL1. The dose-response curves for lactate production and DG uptake were similar. Increased DG uptake was blocked by specific antisera to IL1 and by inhibitors of protein and RNA synthesis but not by indomethacin, and inhibitor of prostaglandin production. DG uptake was enhanced by IL1 in serum-starved cells in the presence of neutralizing anti-platelet-derived growth factor serum. The effect was therefore not secondary to prostaglandin or platelet-derived growth factor production. No increase in cell cycling was detected in IL1-treated cells under the experimental conditions. Kinetic analysis revealed that the Vmax for DG uptake was increased by IL1 (from 36 to 144 pmol/min/mg of cell protein), whereas the Km was unchanged. HuGi cells were pulse-labeled with [35S]methionine following exposure to IL1. Cell lysates were immunoprecipitated using a specific antiserum raised against human erythrocyte glucose transporter. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/autoradiography of these immunoprecipitates revealed dose- and time-dependent increases in the net rate of glucose transporter synthesis which mirrored the changes in DG uptake.