DUAL FUNCTIONAL-EFFECTS OF INTERLEUKIN-1-BETA ON PURINE NUCLEOTIDES AND INSULIN-SECRETION IN RAT ISLETS AND INS-1 CELLS

Interleukin-1 beta (IL-1 beta) has been shown to inhibit glucose-induc ed insulin secretion from rat islets and purified beta-cells, primaril y through the generation of nitric oxide (NO). However, the mechanisms by which NO exerts its effects remain unclear. To examine the role of purine nucleotides, we cultured intact rat islets or INS-1 (glucose-r esponsive transformed rat) beta-cells for 18 h in the presence or abse nce of IL-1 beta. In islets, the exposure to IL-1 beta (100 pmol/l) in hibited subsequent glucose-induced insulin secretion by 91% with no si gnificant effect on insulin content or basal insulin release. IL-1 bet a also diminished insulin secretion induced by pure mitochondrial fuel s, 40 mmol/l K+, or a phorbol ester. Concomitantly, IL-1 beta signific antly decreased islet ATP (-45%), GTP (-33%), ATP/ADP (-54%), and GTP/ GDP (-46%). These effects mere totally reversed by provision of N-omeg a-nitro-L-arginine methyl ester (NAME) in arginine-free media that inh ibited NO production. In contrast, in INS-1 cells, IL-1 beta (10 or 10 0 pmol/l) reduced both basal and glucose-induced insulin secretion by 50%, but insulin content was also reduced by 35%. Therefore, the INS-1 cells were still able to respond to glucose stimulation with a 1.8-2. 0-fold increase in insulin release in either the presence or absence o f IL-1 beta. Concomitantly, in INS-1 cells, IL-1 beta had no effect on ATP/ADP or GTP/GDP ratios, although it modestly decreased ATP (-25%) and GTP (-22%). As in islets, all effects of IL-1 beta in INS-1 cells were prevented by NAME. Thus, in rat islets, IL-1 beta (via the genera tion of NO) abolishes insulin exocytosis in association with large dec reases in the ATP/ADP (and GTP/GDP) ratio, implying the impairment of mitochondrial function. Furthermore, IL-1 beta inhibits cytosolic synt hesis of new purine nucleotides (via the salvage pathway), as assessed by a decrease in their specific activity after labeling with [H-3]hyp oxanthine. In contrast, in INS-1 cells, IL-1 beta appears to impair cy tosolic synthesis of purine nucleotides and insulin biosynthesis selec tively (both possibly reflecting decreased glycolysis) with little dir ect effect on insulin exocytosis itself.