Previous reports have demonstrated that normal serum can increase the rate of adipocyte lipolysis in vitro. However, the nature of the lipolytic activity has remained obscure. We have investigated the lipolytic activity of human serum using isolated rat adipocytes. Human serum resulted in a dose-dependent stimulation of lipolysis (glycerol release) in adipocytes, with a half-maximal effective dose of 0.05% serum and with maximal stimulation with 1% serum. The effect of serum on glycerol release was rapid (within 30 min), and the effect was reversible. Partial purification of this lipolytic activity using gel filtration and ion-exchange chromatography demonstrates that a protein of similar to80 kDa contributes to the lipolytic activity. Human transferrin mimicked the activity of partially purified serum, resulting in a maximal 50% increase in basal lipolysis. In addition, ferrous sulfate heptahydrate induced a biphasic increase in the rate of lipolysis, with a maximal increase of 50% at similar to0.6 mug/ml iron. Inhibitors of protein kinase A (1189) and mitogen-activated protein kinase (PD98059) did not block the effect of serum on lipolysis, whereas the free radical scavenger N-acetyl-L-cysteine completely inhibited the effect. These findings suggest that the stimulatory effect of serum on lipolysis is in part mediated by iron, probably through a prooxidant mechanism.
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