, Volume 36, Issue 1, pp 9–16 | Cite as

Interleukin-1β effects on cyclic GMP and cyclic AMP in cultured rat islets of Langerhans — arginine — dependence and relationship to insulin secretion

  • I. C. Green
  • C. A. Delaney
  • J. M. Cunningham
  • V. Karmiris
  • C. Southern


When islets were cultured with interleukin-1β (1 or 100 pmol/l) for 12 h in arginine-containing medium, cyclic GMP levels were increased 1.6- and 4.5-fold respectively. The arginine analogue, N-ω-nitro-l-arginine methyl ester, which blocks nitric oxide formation and partially reverses inhibition of insulin secretion by 100 pmol/l interleukin-1β, largely, but not completely, blocked generation of cyclic GMP. Treatment of islets with 100 pmol/l interleukin-1β for 12 h significantly decreased islet cyclic AMP generation in the absence of isobutylmethylxanthine (from 13.1±0.7 to 9.3±0.8 fmol/μg islet protein), this fall was arginine-dependent and may have resulted from an effect on a cyclic AMP phosphodiesterase, since it was masked if isobutylmethylxanthine was present. Isobutylmethylxanthine (0.4 mmol/l) reduced the inhibitory potency of interleukin-1β in 15 h slightly but significantly from 80.5 to 59.0%. The morpholinosydnonimine SIN-1, which is a nitric oxide donor, inhibited insulin secretion, raised islet cyclic GMP and lowered cyclic AMP; its effects were similar to those of interleukin-1β. However, 6-anilinoquinoline-5,8-quinone, [LY83583 (1–10 μmol/l)], inhibited insulin secretion, and significantly decreased cyclic GMP while 8-bromocyclic GMP stimulated insulin secretion. Both low- and high-dose interleukin-1β treatment give a large arginine-dependent and a small, yet significant, arginine-independent increase in cyclic GMP. The inhibitory effect of SIN-1 or interleukin-1β on insulin secretion seems to depend to a small extent on decreased islet cyclic AMP, though sustained increases in nitric oxide or depleted islet GTP may directly affect the secretory process.

Key words

Cyclic GMP cyclic AMP interleukin-1β LY83 583 SIN-1 8-bromo cGMP 


  1. 1.
    Mandrup-Poulsen T, Helqvist S, Wogensen LD et al. (1990) Cytokines and free radicals as effector molecules in the destruction of pancreatic β-cells. In: Baekkeshov S, Hansen B (eds) Human diabetes — genetic, environmental and autoimmune etiology. Springer, Berlin Heidelberg New York, pp 169–193Google Scholar
  2. 2.
    Rabinovitch A, Sumoski W, Rajotte RV, Warnock G (1990) Cytotoxic effects of cytokines on human pancreatic islet cells in monolayer culture. J Clin Endocrinol Metab 71: 152–156PubMedGoogle Scholar
  3. 3.
    Spinas GA, Mandrup-Poulsen T, Molvig J et al. (1986) Low concentrations of interleukin-1 stimulate and high concentrations inhibit insulin release from isolated rat islets of Langerhans. Acta Endocrinol (Copenh) 113: 551–558Google Scholar
  4. 4.
    Comens P, Wolf BA, Unanue ER, Lacy PE, McDaniel ML (1987) Interleukin 1 is a potent modulator of insulin secretion from isolated rat islets of Langerhans. Diabetes 36: 963–970PubMedGoogle Scholar
  5. 5.
    Johannesen J, Helqvist S, Nerup J (1990) Not all insulin secretagogues sensitize pancreatic islets to recombinant interleukin 1β. Acta Endocrinol (Copenh) 123: 445–452Google Scholar
  6. 6.
    Zawalich WS, Zawalich KC, Rasmussen H (1989) Interleukin 1a exerts glucose-dependent stimulatory and inhibitory effects on islet cell phosphoinositide hydrolysis and insulin secretion. Endocrinology 124: 2350–2357PubMedGoogle Scholar
  7. 7.
    Eizirik DL, Bendtzen K, Sandler S (1991) Short exposure of rat pancreatic islets to IL-1β induces a sustained but reversible impairment in β-cell function: influence of protease activation, gene transcription and protein synthesis. Endocrinology 128: 1611–1616PubMedGoogle Scholar
  8. 8.
    Sandler S, Bendtzen K, Eizirik DL, Sjoholm Å, Welsh N (1989) Decreased cell replication and polyamine content in insulin-producing cells after exposure to human interleukin-1. Immunol Lett 22: 267–272CrossRefPubMedGoogle Scholar
  9. 9.
    Eizirik DL, Sandler S, Hallberg A, Bendtzen K, Sener A, Malaisse WJ (1989) Differential sensitivity to β-cell secretagogues in cultured rat pancreatic islets exposed to human interleukin-1β. Endocrinology 125: 752–759PubMedGoogle Scholar
  10. 10.
    Welsh N, Sandler S (1992) Interleukin-1β induces nitric oxide production and inhibits the activity of aconitase without decreasing glucose oxidation rates in isolated mouse pancreatic islets. Biochim Biophys Res Commun 182: 333–340Google Scholar
  11. 11.
    Rabinovitch A, Baquerizo H, Sumoski W (1990) Cytotoxic effects of cytokines on islet β-cells: evidence for involvement of eicosanoids. Endocrinology 126: 67–71PubMedGoogle Scholar
  12. 12.
    Bergmann L, Kroncke K-D, Suschek C, Kolb H, Kolb-Bachofen V (1992) Cytotoxic action of IL-1β against pancreatic islets is mediated via nitric oxide formation and is inhibited by N-G-monomethyl-l-rginine. FEBS Lett 299: 103–106CrossRefPubMedGoogle Scholar
  13. 13.
    Hughes JH, Easom RA, Wolf BA, Turk J, McDaniel M (1989) Interleukin 1-duced prostaglandin E2 accumulation by isolated pancreatic islets. Diabetes 38: 1251–1257PubMedGoogle Scholar
  14. 14.
    Welsh N, Nilsson T, Hallberg A, Arkhammar P, Berggren P-O, Sandler S (1989) Human interleukin-1β stimulates islet insulin release by a mechanism not dependent on changes in phospholipase C, protein kinase C activities or Ca2+ handling. Acta Endocrinol (Copenh) 121: 698–704Google Scholar
  15. 15.
    Beggs M, Beresford G, Clarke J, Metz R, Espinal J, Hammonds P (1990) Interleukin-1β inhibits glucokinase activity in clonal HIT-T15 β-cells. FEBS Lett 267: 217–220CrossRefPubMedGoogle Scholar
  16. 16.
    Sumoski W, Baquerizo H, Rabinovitch A (1989) Oxygen free radical scavengers protect rat islet cells from damage by cytokines. Diabetologia 32: 792–796CrossRefPubMedGoogle Scholar
  17. 17.
    Sjöholm Å (1991) Inhibition of fetal rat pancreatic β-cell replication by interleukin-1β in vitro is not mediated through pertussis toxin-sensitive G-proteins, a decrease in cyclic AMP or protease activation. FEBS Lett 289: 249–252CrossRefPubMedGoogle Scholar
  18. 18.
    Borg LAH, Sandler S, Eizirik DL (1991) Interleukin-1β increases the activity of superoxide dismutase in rat pancreatic islets. Diabetologia 34 [Suppl 2]: A8 (Abstract)Google Scholar
  19. 19.
    Welsh N, Bendtzen K, Sandler S (1991) Influence of protease on inhibitory and stimulatory effects of interleukin 1β on β-cell function. Diabetes 40: 290–294PubMedGoogle Scholar
  20. 20.
    Helqvist S, Hansen BS, Johannesen J, Andersen HU, Nielsen JH, Nerup J (1989) Interleukin 1 induces new protein formation in isolated rat islets of Langerhans. Acta Endocrinol (Copenh) 121: 136–140Google Scholar
  21. 21.
    Helqvist S, Polla BS, Johannesen J, Nerup J (1991) Heat shock protein induction in rat pancreatic islets by recombinant human interleukin 1β. Diabetologia 34: 150–156PubMedGoogle Scholar
  22. 22.
    Hughes JH, Colca J, Easom RA, Turk J, McDaniel ML (1990) In-terleukin 1 inhibits insulin secretion from isolated rat pancreatic islets by a process that requires gene transcription and mRNA translation. J Clin Invest 86: 856–863PubMedGoogle Scholar
  23. 23.
    Eizirik DL, Hallgren IB, Forsbeck E, Sandler S (1991) Interleukin-1β actions on β-cells are mediated by type 1 IL-1 receptors and gene transcription. Diabetologia 34 [Suppl 2]: A9 (Abstract)Google Scholar
  24. 24.
    Southern C, Schulster D, Green IC (1990) Inhibition of insulin secretion by interleukin-1β and tumour necrosis factor-α via an l-arginine-dependent nitric oxide generating mechanism. FEBS Lett 276: 42–44CrossRefPubMedGoogle Scholar
  25. 25.
    Corbett JA, Lancaster ] Jr, Sweetland MA, McDaniel ML (1991) Interleukin-1β-induced formation of EPR-detectable iron-nitrosyl complexes in islets of Langerhans. J Biol Chem 266: 21351–21354PubMedGoogle Scholar
  26. 26.
    Kroncke KD, Kolb-Bachofen V, Berschick B, Burkart V, Kolb H (1991) Activated macrophages kill pancreatic syngeneic islet cells via arginine-dependent nitric oxide generation. Biochim Biophys Res Commun 175: 752–758Google Scholar
  27. 27.
    Jansson L, Sandler S (1991) The nitric oxide synthase II inhibitor N-nitro-l-arginine stimulates pancreatic islet insulin release in vitro, but not in the perfused pancreas. Endocrinology 128: 3081–3085PubMedGoogle Scholar
  28. 28.
    Moncada S, Palmer RMJ, Higgs EA (1991) Nitric oxide: physiology, pathophysiology and pharmacology. Pharmacol Rev 43: 109–142PubMedGoogle Scholar
  29. 29.
    Waldman SA, Murad F (1987) Cyclic GMP synthesis and function. Pharmacol Rev 39: 163–196PubMedGoogle Scholar
  30. 30.
    Sandler S, Bendtzen K, Eizirik DL, Strandell E, Welsh M, Welsh N (1990) Metabolism and B-cell function of rat pancreatic islets exposed to human interleukin-1β in the presence of a high glucose concentration. Immunol Lett 26: 245–252CrossRefPubMedGoogle Scholar
  31. 31.
    Gey GO, Gey MH (1936) Maintenance of human normal cells and tumor cells in continuous culture. Am J Cancer 27: 45–76Google Scholar
  32. 32.
    Green IC, Perrin D, Penman E, Ray K, Howell SL (1983) Effect of dynorphin on insulin and somatostatin secretion, calcium uptake and c-AMP levels in isolated rat islets of Langerhans. Diabetes 32: 685–690PubMedGoogle Scholar
  33. 33.
    Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254CrossRefPubMedGoogle Scholar
  34. 34.
    Brooker G, Harper JF, Terasaki WL, Moylan RD (1979) Radioimmunoassay of cyclic AMP and cyclic GMP. Adv Cyclic Nucleotide Res 10: 1–33PubMedGoogle Scholar
  35. 35.
    Green IC, Ray K, Perrin D (1983) Opioid peptide effects on insulin release and c-AMP in islets of Langerhans. Horm Metab Res 15: 124–128PubMedGoogle Scholar
  36. 36.
    Bogle RG, Moncada S, Pearson JD, Mann GE (1992) Identification of inhibitors of nitric oxide synthase that do not interact with the endothelial cell l-arginine transporter. Br J Pharmacol 105: 768–770PubMedGoogle Scholar
  37. 37.
    Knowles RG, Palacios M, Palmer RMJ, Moncada S (1990) Nitric oxide synthase in the brain. In: Moncada S, Higgs EA (eds) Nitric oxide from l-arginine. A bioregulatory system. Excerpta Medica International, Amsterdam, pp 139–147Google Scholar
  38. 38.
    Stuehr DJ, Cho HJ, Kwon NS, Nathan CF (1992) The cytokineinduced macrophage nitric oxide synthase is an FAD-and FMN-containing flavoprotein. 2nd International Meeting “The Biology of nitric oxide” Part II Enzymology Biochemistry and Immunology, September 1991. Portland Press, LondonGoogle Scholar
  39. 39.
    Laychock SG (1981) Evidence for guanosine 3′5′-monophosphate as a putative mediator of insulin secretion from isolated rat islets. Endocrinology 108: 1197–1205PubMedGoogle Scholar
  40. 40.
    Laychock SG, Modica ME, Cavanaugh CT (1991) l-arginine stimulates cyclic guanosine 3′,5′-monophosphate formation in rat islets of Langerhans and RINm5F insulinoma cells: evidence for l-arginine: nitric oxide synthase. Endocrinology 129: 3043–3052PubMedGoogle Scholar
  41. 41.
    Schmidt HHW, Warner TD, Ishi K, Sheng H, Murad F (1992) Insulin secretion from pancreatic B cells caused by l-arginine derived nitrogen oxides. Science 255: 721–723PubMedGoogle Scholar
  42. 42.
    Jones PM, Persaud SJ, Howell SL (1992) Is cyclic GMP involved in insulin secretion from rat islets? Diabetic Med 9 [Suppl 1]: 12 A (Abstract)Google Scholar
  43. 43.
    Southam E, Garthwaite J (1991) Comparative effects of some nitric oxide donors on cyclic GMP levels in rat cerebellar slices. Neurosci Lett 130: 107–111CrossRefPubMedGoogle Scholar
  44. 44.
    Gross SS, Levi R (1992) Synthesis of tetrahydrobiopterin is a requirement for induction of nitric oxide synthesis by LPS/interferon in vascular smooth muscle. In: 2nd International Meeting “Biology of nitric oxide” September 1991. Portland Press, LondonGoogle Scholar
  45. 45.
    Metz SA, Rabaglia M, Pintar TJ (1992) Selective inhibitors of GTP synthesis impede exocytotic insulin release from intact rat islets. J Biol Chem 267: 12517–12527PubMedGoogle Scholar
  46. 46.
    Terry BJ, Purich DK (1980) Assembly and disassembly properties of microtubules formed in the presence of GTP, 5′ guanylyl imidodiphosphate, and 5′-guanylyl methylenediphosphate. J Biol Chem 255: 10532–10536PubMedGoogle Scholar
  47. 47.
    Iyengar R, Abramowitz J, Bordelon-Riser M et al. (1980) Hormone receptor-mediated stimulation of adenylyl cyclase systems. Nucleotide effects and analysis in terms of a simple two state-model for the basic receptor-affected enzyme. J Biol Chem 255: 3558–3564PubMedGoogle Scholar
  48. 48.
    Meglasson MD, Nelson J, Nelson D, Erecinska M (1989) Bioenergetic response of pancreatic islets to stimulation by fuel molecules. Metabolism 38: 1188–1195CrossRefPubMedGoogle Scholar
  49. 49.
    Hoenig M, Matschinsky FM (1987) HPLC analysis of nucleotide profiles in glucose-stimulated perifused rat islets. Metabolism 36: 295–301CrossRefPubMedGoogle Scholar
  50. 50.
    Howell SL, Green IC, Montague W (1973) A possible role of adenylate cyclase in the long term dietary regulation of insulin secretion from rat islets of Langerhans. Biochem J 136: 343–349PubMedGoogle Scholar
  51. 51.
    Mizel SB (1990) Cyclic AMP and interleukin 1 signal transduction. Immunl Today 11: 390–394CrossRefGoogle Scholar
  52. 52.
    MacFarland RT, Zelus BD, Beavo J (1991) High concentrations of a cyclic GMP-stimulated phosphodiesterase mediate ANP-induced decreases in cAMP and steroidogenesis in adrenal glomerulosa cells. J Biol Chem 266: 136–142PubMedGoogle Scholar
  53. 53.
    Stroop SD, Beavo JA (1991) Structure and function studies of the cGMP-stimulated phosphodiesterase. J Biol Chem 266: 23802–23809PubMedGoogle Scholar
  54. 54.
    Hammonds P, Beggs M, Beresford G, Espinal J, Clarke J, Metz RJ (1990) Insulin-secretory β-cells possess specific receptors for interleukin-1β. FEBS Lett 261: 97–100CrossRefPubMedGoogle Scholar
  55. 55.
    Furman BL, Pyne NJ (1990) Islet phosphodiesterase isoenzymes and insulin secretion. Diabetic Med 7 [Suppl 1]: 19 A (Abstract)Google Scholar
  56. 56.
    Beavo JA, Reifsnyder DH (1990) Primary sequence of cyclic nucleotide phosphodiesterase isozymes and the design of selective inhibitors. TiPS 11: 150–155PubMedGoogle Scholar
  57. 57.
    Southern C, Schulster D, Green IC (1991) Nitric-oxide stimulated cGMP, relationship to cytokine effects on insulin secretion. Diabetic Med 8 [Suppl 1]: 1A (Abstract)PubMedGoogle Scholar
  58. 58.
    Corbett JA, Tilton RA, Chang K et al. (1992) Aminoguanidine, a novel inhibitor of nitric oxide formation, prevents diabetic vascular dysfunction. Diabetes 41: 552–556PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • I. C. Green
    • 1
  • C. A. Delaney
    • 1
  • J. M. Cunningham
    • 1
  • V. Karmiris
    • 1
  • C. Southern
    • 2
  1. 1.Biochemistry LaboratorySchool of Biological Sciences University of SussexBrighton
  2. 2.Yamanouchi Research InstituteLittlemore HospitalOxfordUK

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