Molecular Medicine

, Volume 9, Issue 3–4, pp 96–104 | Cite as

Inosine Protects Against the Development of Diabetes in Multiple-Low-Dose Streptozotocin and Nonobese Diabetic Mouse Models of Type 1 Diabetes

  • Jon G Mabley
  • Alex Rabinovitch
  • Wilma Suarez-Pinzon
  • György Haskó
  • Pál Pacher
  • Robert Power
  • Gary Southan
  • Andrew Salzman
  • Csaba Szabó


Inosine, a naturally occurring purine, was long considered to be an inactive metabolite of adenosine. However, recently inosine has been shown to be an immunomodulator and anti-inflammatory agent. The aim of this study was to determine whether inosine influences anti-inflammatory effects and affects the development of type 1 diabetes in murine models. Type 1 diabetes was induced either chemically by streptozotocin or genetically using the nonobese diabetic mouse (NOD) model. Mice were treated with inosine (100 or 200 mg kg1d1) and diabetes incidence was monitored. The effect of inosine on pancreas immune cell infiltration, oxidative stress, and cytokine profile also was determined. For the transplantation model islets were placed under the renal capsule of NOD mice and inosine (200 mg kg1d1) treatment started the day of islet transplantation. Graft rejection was diagnosed by return of hyperglycemia accompanied by glucosuria and ketonuria. Inosine reduced the incidence of diabetes in both streptozotocin-induced diabetes and spontaneous diabetes in NOD mice. Inosine decreased pancreatic leukocyte infiltration and oxidative stress in addition to switching the cytokine profile from a Th1 to a Th2 profile. Inosine prolonged pancreatic islet graft survival, increased the number of surviving β cells, and reduced the number of infiltrating leukocytes. Inosine protects against both the development of diabetes and against the rejection of transplanted islets. The purine exerts anti-inflammatory effects in the pancreas, which is its likely mode of action. The use of inosine should be considered as a potential preventative therapy in humans susceptible to developing Type 1 diabetes and as a possible antirejection therapy for islet transplant recipients.



This study was supported by grants from the National Institutes of Health (1R43 DK59676 to GS) and the Canadian Institutes of Health Research (1-199-908 to AR).


  1. 1.
    Cronstein BN. (1994) Adenosine, an endogenous anti-inflammatory agent. J. Appl. Physiol. 76:5–13.CrossRefGoogle Scholar
  2. 2.
    Apasov S, Koshiba M, Redegeld F, Sitkovsky MV. (1995) Role of extracellular ATP and P1 and P2 classes of purinergic receptors in T-cell development and cytotoxic T lymphocyte effector functions. Immunol. Rev. 146:5–19.CrossRefGoogle Scholar
  3. 3.
    Hasko G, Szabo C. (1998) Regulation of cytokine and chemokine production by transmitters and co-transmitters of the autonomic nervous system. Biochem. Pharmacol. 56:1079–87.CrossRefGoogle Scholar
  4. 4.
    Hasko G, Nemeth ZH, Vizi ES, Salzman AL, Szabo C. (1998) An agonist of adenosine A3 receptors decreases interleukin-12 and interferon-γ production and prevents lethality in endotoxemic mice. Eur. J. Pharmacol. 358:261–8.CrossRefGoogle Scholar
  5. 5.
    Szabo C et al. (1998) Suppression of macrophage inflammatory protein (MIP)-1α production and collagen-induced arthritis by adenosine receptor agonists. Br. J. Pharmacol. 125:379–87.CrossRefGoogle Scholar
  6. 6.
    Schrier DJ, Lesch ME, Wright CD, Gilbertsen RB. (1990) The antiinflammatory effects of adenosine receptor agonists on the carrageenan-induced pleural inflammatory response in rats. J. Immunol. 145:1874–9.PubMedGoogle Scholar
  7. 7.
    Poelstra K, Heynen ER, Baller JF, Hardonk MJ, Bakker WW. (1992) Modulation of anti-Thy1 nephritis in the rat by adenine nucleotides. Evidence for an anti-inflammatory role for nucleotidases. Lab. Invest. 66:555–63.PubMedGoogle Scholar
  8. 8.
    Marak Jr GE et al. (1988) Pharmacologic modulation of acute ocular inflammation. I. Adenosine. Ophthalmic Res. 20:220–6.CrossRefGoogle Scholar
  9. 9.
    Mabley JG et al. (2003) Inosine reduces inflammation and improves survival in a murine model of colitis. Am. J. Physiol. Gastrointest. Liver Physiol. 284:G138–44.CrossRefGoogle Scholar
  10. 10.
    Hasko G et al. (2000) Adenosine inhibits IL-12 and TNF-α production via adenosine A2a receptor-dependent and independent mechanisms. FASEB J. 14:2065–74.CrossRefGoogle Scholar
  11. 11.
    Barankiewicz J, Cohen A. (1985) Purine nucleotide metabolism in resident and activated rat macrophages in vitro. Eur. J. Immunol. 15:627–31.CrossRefGoogle Scholar
  12. 12.
    Hasko G et al. (2000) Inosine inhibits inflammatory cytokine production by a posttranscriptional mechanism and protects against endotoxin-induced shock. J. Immunol. 164:1013–9.CrossRefGoogle Scholar
  13. 13.
    Garcia Soriano F et al. (2001) Inosine improves gut permeability and vascular reactivity in endotoxic shock. Crit. Care Med. 29:703–8.CrossRefGoogle Scholar
  14. 14.
    Liaudet L et al. (2001) Inosine reduces systemic inflammation and improves survival in septic shock induced by cecal ligation and puncture. Am. J. Respir. Crit. Care Med. 164:1213–20.CrossRefGoogle Scholar
  15. 15.
    Liaudet L et al. (2002) Inosine exerts a broad range of anti-inflammatory effects in a murine model of acute lung injury. Ann. Surgery 235:568–78.CrossRefGoogle Scholar
  16. 16.
    Marton A et al. (2001) Anti-inflammatory effects of inosine in human monocytes, neutrophils and epithelial cells in vitro. Int. J. Mol. Med. 8:617–21.PubMedGoogle Scholar
  17. 17.
    Bach J-F. (1994) Insulin-dependent diabetes mellitus as an autoimmune disease. Endocr. Rev. 15:516–42.CrossRefGoogle Scholar
  18. 18.
    Rabinovitch A, Suarez-Pinzon WL. (1998) Cytokines and their roles in pancreatic islet β-cell destruction and insulin-dependent diabetes mellitus. Biochem. Pharmacol. 55:1139–49.CrossRefGoogle Scholar
  19. 19.
    Rabinovitch A. (1998) An update on cytokines in the pathogenesis of insulin-dependent diabetes mellitus. Diabetes Metab. Rev. 14:129–51.CrossRefGoogle Scholar
  20. 20.
    Like AA, Rossini AA. (1976) Streptozotocin-induced pancreatic insulitis: new model of diabetes mellitus. Science 193:415–7.CrossRefGoogle Scholar
  21. 21.
    Rossini AA, Williams RM, Appel MC, Like AA. (1978) Complete protection from low-dose streptozotocin-induced diabetes in mice. Nature 276:182–4.CrossRefGoogle Scholar
  22. 22.
    Atkinson MA, Leiter EH. (1999) The NOD mouse model of type 1 diabetes: as good as it gets? Nat. Med. 5:601–4.CrossRefGoogle Scholar
  23. 23.
    Mabley JG et al. (2001) Inhibition of poly (ADP-ribose) synthetase by gene disruption or inhibition with 5-iodo-6-amino-1,2-benzopyrone protects mice from multiple-low-dose-streptozotocin-induced diabetes. Br. J. Pharmacol. 133:909–19.CrossRefGoogle Scholar
  24. 24.
    Chatenoud L, Primo J, Bach JF. (1997) CD3 antibody-induced dominant self tolerance in overtly diabetic NOD mice. J. Immunol. 158:2947–54.PubMedGoogle Scholar
  25. 25.
    Ryu S, Kodama S, Ryu K, Schoenfeld DA, Faustman DL. (2001) Reversal of established autoimmune diabetes by restoration of endogenous beta cell function. J. Clin. Invest. 108:63–72.CrossRefGoogle Scholar
  26. 26.
    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. 72:248–54.CrossRefGoogle Scholar
  27. 27.
    Mabley JG, Virag L, Szabo C. (2002) Role of poly (ADP-ribose) polymerase activation in the pathogenesis of diabetes mellitus and diabetic vascular dysfunction. In: PARP as a therapeutic target. Zhang J (ed.) CRC Press, London, pp. 277–319.Google Scholar
  28. 28.
    Gotoh M, Maki T, Kiyoizumi T, Satomi S, Monaco AP. (1985) An improved method for isolation of mouse pancreatic islets. Transplantation 40:437–8.CrossRefGoogle Scholar
  29. 29.
    Wang T, Singh B, Warnock GL, Rajotte RV. (1992) Prevention of recurrence of IDDM in islet-transplanted diabetic NOD mice by adjuvant immunotherapy. Diabetes 41:114–7.CrossRefGoogle Scholar
  30. 30.
    Suarez-Pinzon WL, Mabley JG, Strynadka K, Power RF, Szabo C, Rabinovitch A. (2001) An inhibitor of inducible nitric oxide synthase and scavenger of peroxynitrite prevents diabetes development in nod mice. J. Autoimmun. 16:449–55.CrossRefGoogle Scholar
  31. 31.
    Eizirik DL, Darville MI. (2001) β-cell apoptosis and defense mechanisms: lessons from type 1 diabetes. Diabetes 50 Suppl 1:S64–9.CrossRefGoogle Scholar
  32. 32.
    Rabinovitch A, Suarez-Pinzon WL, Sorensen O. (1996) Interleukin 12 mRNA expression in islets correlates with β-cell destruction in NOD mice. J. Autoimmun. 9:645–51.CrossRefGoogle Scholar
  33. 33.
    Rothe H, O’Hara Jr RM, Martin S, Kolb H. (1997) Suppression of cyclophosphamide induced diabetes development and pancreatic Th1 reactivity in NOD mice treated with the interleukin (IL)-12 antagonist IL-12(p40)2. Diabetologia 40:641–6.CrossRefGoogle Scholar
  34. 34.
    Kolb H et al. (1987) Analysis of 22 immunomodulatory substances for efficacy in low-dose streptozotocin-induced diabetes. Diabetes Res. 6:21–7.PubMedGoogle Scholar
  35. 35.
    Wybran J, Famaey JP, Appelboom T. (1981) Inosiplex: a novel treatment in rheumatoid arthritis? J. Rheumatol. 8:643–6.PubMedGoogle Scholar
  36. 36.
    Campoli-Richards DM, Sorkin EM, Heel RC. (1986) Inosine pranobex. A preliminary review of its pharmacodynamic and pharmacokinetic properties, and therapeutic efficacy. Drugs 32:383–424.CrossRefGoogle Scholar
  37. 37.
    Ward SG, Bacon K, Westwick J. (1998) Chemokines and T lymphocytes: more than an attraction. Immunity 9:1–11.CrossRefGoogle Scholar
  38. 38.
    Cameron MJ et al. (2000) Differential expression of CC chemokines and the CCR5 receptor in the pancreas is associated with progression to type I diabetes. J. Immunol. 165:1102–10.CrossRefGoogle Scholar
  39. 39.
    Ohta A, Sitkovsky M. (2001) Role of G-protein-coupled adenosine receptors in downregulation of inflammation and protection from tissue damage. Nature 414:916–20.CrossRefGoogle Scholar
  40. 40.
    McWhinney CD et al. (1996) Activation of adenosine A3 receptors on macrophages inhibits tumor necrosis factor-α. Eur. J. Pharmacol. 310:209–16.CrossRefGoogle Scholar
  41. 41.
    Shapira L, Houri Y, Barak V, Soskolne WA, Halabi A, Stabholz A. (1997) Tetracycline inhibits Porphyromonas gingivalis lipopolysaccharide-induced lesions in vivo and TNF-α processing in vitro. J. Periodontal Res. 32:183–8.CrossRefGoogle Scholar
  42. 42.
    Mohler KM et al. (1994) Protection against a lethal dose of endotoxin by an inhibitor of tumour necrosis factor processing. Nature 370:218–20.CrossRefGoogle Scholar
  43. 43.
    Zhang M et al. (1997) Spermine inhibits proinflammatory cytokine synthesis in human mononuclear cells: a counterregulatory mechanism that restrains the immune response. J. Exp. Med. 185:1759–68.CrossRefGoogle Scholar
  44. 44.
    Virag L, Szabo C. (2001) Purines inhibit poly(ADP-ribose) polymerase activation and modulate oxidant-induced cell death. FASEB J 15:99–107.CrossRefGoogle Scholar
  45. 45.
    Mabley JG et al. (2001) Anti-inflammatory effects of a novel, potent inhibitor of poly (ADP-ribose) polymerase. Inflamm. Res. 50:561–9.CrossRefGoogle Scholar
  46. 46.
    Ryan EA et al. (2001) Clinical outcomes and insulin secretion after islet transplantation with the Edmonton protocol. Diabetes 50:710–9.CrossRefGoogle Scholar
  47. 47.
    Jurkowitz MS, Litsky ML, Browning MJ, Hohl CM. (1998) Adenosine, inosine, and guanosine protect glial cells during glucose deprivation and mitochondrial inhibition: correlation between protection and ATP preservation. J. Neurochem. 71: 535–48.CrossRefGoogle Scholar
  48. 48.
    Cole AW, Palmer TN. (1979) Action of purine nucleosides on the release of intra-cellular enzymes from rat lymphocytes. Clin. Chim. Acta. 92:93–100.CrossRefGoogle Scholar
  49. 49.
    de Rougemont D, Brunner FP, Torhorst J, Wunderlich PF, Thiel G. (1982) Superficial nephron obstruction and medullary congestion after ischemic injury: effect of protective treatments. Nephron 31:310–20.CrossRefGoogle Scholar
  50. 50.
    Tilser I, Martinkova J, Chladek J. (1993) The effect of metipranolol and inosine on total hepatic ischemia of rats in vivo. Sb Ved Pr Lek Fak Karlovy Univerzity Hradci Kralove 36:25–9.PubMedGoogle Scholar
  51. 51.
    Scott GS, Spitsin SV, Kean RB, Mikheeva T, Koprowski H, Hooper DC. (2002) Therapeutic intervention in experimental allergic encephalomyelitis by administration of uric acid precursors. Proc. Natl. Acad. Sci. U.S.A. 99:16303–8.CrossRefGoogle Scholar
  52. 52.
    Scott GS, Hooper DC. (2001). The role of uric acid in protection against peroxynitrite-mediated pathology. Med. Hypotheses. 56:95–100.CrossRefGoogle Scholar
  53. 53.
    Williams MH et al. (1990) Effect of inosine supplementation on 3-mile treadmill run performance and VO2 peak. Med. Sci. Sports Exerc. 22:517–22.CrossRefGoogle Scholar
  54. 54.
    Spitsin S et al. (2001) Inactivation of peroxynitrite in multiple sclerosis patients after oral administration of inosine may suggest possible approaches to therapy of the disease. Mult. Scler. 7:313–9.CrossRefGoogle Scholar

Copyright information

© Feinstein Institute for Medical Research 2003

Authors and Affiliations

  • Jon G Mabley
    • 1
  • Alex Rabinovitch
    • 2
  • Wilma Suarez-Pinzon
    • 2
  • György Haskó
    • 3
  • Pál Pacher
    • 1
  • Robert Power
    • 4
  • Gary Southan
    • 1
  • Andrew Salzman
    • 1
  • Csaba Szabó
    • 1
    • 3
  1. 1.Inotek Pharmaceuticals CorpBeverlyUSA
  2. 2.Department of MedicineUniversity of AlbertaEdmontonCanada
  3. 3.Department of SurgeryNew Jersey Medical SchoolNewarkUSA
  4. 4.Department of Laboratory MedicineUniversity of AlbertaEdmontonCanada

Personalised recommendations