Antilipolytic effect of prostaglandin E2 in perifused rat adipocytes.

Abstract

Previously, the antilipolytic effect of prostaglandin E2 (PGE2) has been investigated in conventional adipocyte incubations. To define the effect of PGE2 on lipolysis more clearly, isolated epididymal adipocytes were studied with the perifusion system. PGE2 inhibited isoproterenol (100 nM)- and theophylline (1 mM)-stimulated lipolysis in a concentration-dependent manner in both the perifusion system and conventional incubations. However, the half-maximally inhibitory concentration (ED50) of PGE2 on isoproterenol-induced lipolysis was about 0.4 nM in the perifusion system, whereas the ED50 was 8 nM in the static adipocyte incubations. The ED50 values of PGE2 on theophylline-induced lipolysis were 0.8 nM (perifusion) and 5 nM (incubation), respectively. Thus, the sensitivity of stimulated lipolysis to PGE2 was about 10 times higher in the perifusion system than in conventional adipocyte incubations. In addition, the maximal antilipolytic effect of PGE2 was greater in the perifusion system. At a concentration of 100 nM PGE2 inhibited theophylline-induced lipolysis by 82 +/- 5% in adipocyte incubations, whereas lipolysis was inhibited by 100 +/- 3.5% in the perifusion system (P less than 0.05). When lipolysis was stimulated by isoproterenol the maximal antilipolytic effect of PGE2 was an inhibition of 90 +/- 2.5% in the perifusion system and 55 +/- 5% in adipocyte incubations (P less than 0.05). Moreover, the maximal antilipolytic effect was obtained at a PGE2 concentration of 20 nM in the perifusion system, but at a concentration of 100 nM in static incubations. The release of immunoreactive PGE2 from adipocytes was measured by RIA. In the perifusion system no PGE2 could be detected in the effluent under basal conditions; however, during exposure to 100 nM isoproterenol a small amount of PGE2 was detected (3-4.5 pg/10(6) cells X min). Exogenous PGE2 was almost totally (90%) recovered in the effluent. In adipocyte incubations basal PGE2 production was 103 +/- 22 pg/10(6) cells X 60 min, whereas both isoproterenol and theophylline increased these amounts of PGE2 2-fold (P less than 0.01). It is concluded that exogenous PGE2 has pronounced antilipolytic properties at very low concentrations (subnanomolar) in perifused adipocytes. The reduced sensitivity and maximal responsiveness of PGE2 in static incubations may be related to accumulation of FFA and endogenous PGs, which may partially obscure the interaction of exogenous PGE2 with the adenylate cyclase complex.