Insulin-mediated antilipolysis in permeabilized rat adipocytes

Elucidating the mechanism by which insulin inhibits lipolysis has been hampered by the unavailability of a broken cell preparation in which the intact cell responses to the hormone could be duplicated. Here we report, using digitonin-permeabilized rat adipocytes, that physiological concentrations of insulin inhibit cyclic AMP-activated lipolysis despite the absence of cytosolic and plasma membrane integrity. Cyclic AMP (1.0 mM) maximally activates lipolysis in permeabilized adipocytes greater than 10-fold. Insulin inhibits this activation in a biphasic manner with maximum inhibition of 59 +/- 8% (N = 7) at 10(-9) M. At the submaximal concentrations of cyclic AMP (1.0 to 10 microM), insulin (10(-9) M) inhibits lipolysis 80 to 90%. Additionally, the antilipolytic effect of insulin is rapid (3 min) and it is specific, with the relatively inactive desoctapeptide analogue of insulin being three orders of magnitude less inhibitory than native insulin. In contrast to permeabilized cells, intact cells demonstrate only a small lipolytic response to cyclic AMP which is insensitive to insulin. These findings suggest the following about insulin's antilipolytic effects: 1) an intact cell is not required; 2) the intracellular mechanism of action does not require physiological concentrations of the freely diffusible cytosolic components; and 3) a site of insulin action independent of adenylate cyclase may play a major role.