Tumor necrosis factor-alpha and ceramides in insulin resistance

The present studies tested the hypothesis that some effects of tumor necros is factor-alpha (TNF-alpha) are mediated by activation of sphingomyelinases and the production of ceramides. Differentiated 3T3-L1 adipocytes were inc ubated with short-chain ceramide analogs, (C-2- and C-6-ceramides: N-acetyl - and N-hexanoyl-sphingosines, respectively), and this treatment increased 2-deoxyglucose uptake in the absence of insulin progressively from 2-24 h. This effect was inhibited by blocking the activations of mitogen-activated protein kinase, phosphatidylinositol 3-kinase (PI 3-kinase), and ribosomal S6 kinase which mediated an increase in GLUT1 concentrations. Long-term inc reases in PI 3-kinase activity associated with insulin receptor substrate-1 (IRS-1) increased the proportion of GLUT1 and GLUT4 in plasma membranes. T hese events explain the increases in noninsulin-dependent glucose uptake an d incorporation of this glucose into the fatty acid and glycerol moieties o f triacylglycerol. The mechanisms by which TNF-alpha and ceramides increase PI 3-kinase activity were investigated further by using rat2 fibroblasts. Incubation for 20 min with TNF-alpha, bacterial sphingomyelinase, or C-2-ce ramides increased Pt 3-kinase activity by about fivefold, and this effect d epended upon a stimulation of tyrosine kinase activity and an increase in R as-GTP. This demonstrates the existence of a novel signaling pathway far TN F-alpha that could contribute to the effects of this cytokine in stimulatin g basal glucose uptake. By contrast, treating the 3T3-L1 adipocytes for 2-2 4 h with C-2-ceramide diminished insulin-stimulated glucose uptake by decre asing the insulin-induced translocation of GLUT1 and GLUT4 to plasma membra nes. This inhibition was observed when there was no increase in basal gluco se uptake, and it occurred downstream of PI 3-kinase. Our work provides fur ther mechanisms whereby TNF-alpha and ceramides produce insulin resistance and decrease the effectiveness of insulin in stimulating glucose disposal f rom the blood. Conversely, TNF-alpha and ceramides increase the ability of adipocytes to take up glucose and store triacylglycerol in the absence of i nsulin.