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J Biol Chem, Vol. 274, Issue 10, 6770-6775, March 5, 1999

Tumor Necrosis Factor  Stimulates Lipolysis in Adipocytes by Decreasing G_i Protein Concentrations

Slavisa Gasic, Bing Tian^§, and Allan Green

From the  Bassett Research Institute, The Mary Imogene Bassett Hospital, Cooperstown, New York 13326 and the ^§ Department of Pharmacology, University of Texas Medical Branch, Galveston, Texas 77555

Prolonged treatment (12-24 h) of adipocytes with tumor necrosis factor  (TNF) stimulates lipolysis. We have investigated the hypothesis that TNF stimulates lipolysis by blocking the action of endogenous adenosine. Adipocytes were incubated for 48 h with TNF, and lipolysis was measured in the absence or presence of adenosine deaminase. Without adenosine deaminase, the rate of glycerol release was 2-3-fold higher in the TNF-treated cells, but with adenosine deaminase lipolysis increased in the controls to approximately that in the TNF-treated cells. This suggests that TNF blocks adenosine release or prevents its antilipolytic effect. Both N⁶-phenylisopropyl adenosine and nicotinic acid were less potent and efficacious inhibitors of lipolysis in treated cells. A decrease in the concentration of -subunits of all three G_i subtypes was detected by Western blotting without a change in G_s proteins or -subunits. G_i2 was about 50% of control, whereas G_i1 and G_i3 were about 20 and 40% of control values, respectively. The time course of G_i down-regulation correlated with the stimulation of lipolysis. Furthermore, down-regulation of G_i by an alternative approach (prolonged incubation with N⁶-phenylisopropyl adenosine) stimulated lipolysis. These findings indicate that TNF stimulates lipolysis by blunting endogenous inhibition of lipolysis. The mechanism appears to be a G_i protein down-regulation.

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