Summary
Lysis of rat islet cells by syngeneic activated macrophages in vitro can be completely inhibited by the nitric oxide-synthase-inhibitor NG-methyl-l-arginine. This inhibition can be reversed by an excess of l-arginine. Time-dependent lysis of islet cells by activated macrophages is accompanied by increasing concentrations of nitrite and citrulline in the culture medium both of which are measures of nitric oxide formation derived from l-arginine. Lysis of isolated islet cells and disintegration of isolated whole islets is also obtained within 15 h by culture in the presence of the nitric oxide generating vasodilator sodium nitroprusside. We thus conclude that nitric oxide is extremely toxic for islet cells and that nitric oxide alone and in the absence of other macrophage-generated potentially toxic products can rapidly and completely kill islet cells.
Article PDF
Similar content being viewed by others
References
Kolb-Bachofen V, Epstein S, Kiesel U, Kolb H (1988) Low dose streptozotocin-induced diabetes in mice. Electron microscopy reveals single-cell insulitis before diabetes onset. Diabetes 37: 21–27
Lee KU, Kim MK, Amano K et al. (1988) Preferential infiltration of macrophages during early stages of insulitis in diabetesprone BB rats. Diabetes 37: 1053–1058
Hanenberg H, Kolb-Bachofen V, Kantwerk-Funke G, Kolb H (1989) Macrophage infiltration precedes and is pre-requisite for lymphocytic insulitis of prediabetic BB rats. Diabetologia 32: 126–134
Oschilewksi M, Kiesel U, Kolb H (1986) Administration of silica prevents diabetes in BB rats. Diabetes 34: 197–199
Lee KU, Amano K, Yoon J (1988) Evidence for initial involvement of macrophages in development of insulitis in NOD mice. Diabetes 37: 989–991
Marletta MA, Yoon PS, Iyengar R, Leaf CD, Wishnok JS (1988) Macrophage oxidation of l-arginine to nitrite and nitrate: nitric oxide is an intermediate. Biochemistry 27: 8706–8711
Hibbs JB, Taintor RR, Vavrin Z, Rachlin EM (1988) Nitric oxide: a cytotoxic activated macrophage effector molecule. Biochem Biophys Res Commun 157: 87–94
Stuehr DJ, Gross SS, Sakuma I, Levi R, Nathan CF (1989) Activated murine macrophages secrete a metabolite of arginine with the bioactivity of endothelium-derived relaxing factor and the chemical reactivity of nitric oxide. J Exp Med 169: 1011–1020
James SL, Glaven J (1989) Macrophage cytotoxicity against schistosomula of Schistosoma mansoni involves argininedependent production of reactive nitrogen intermediates. J Immunol 143: 4208–4212
Green SJ, Meltzer MS, Hibbs JB, Nacy CA (1990) Activated macrophages destroy intracellular Leishmania major amastigotes by an l-arginine-dependent killing mechanism. J Immunol 144: 278–283
Adams LB, Hibbs JB, Taintor RR, Krahenbuhl JL (1990) Microbiostatic effect of murine-activated macrophages for Toxoplasma gondii. Role for synthesis of inorganic nitrogen oxides from l-arginine. J Immunol 144: 2725–2729
Hibbs JB, Vavrin Z, Taintor RR (1987) l-arginine is required for expression of the activated macrophage effector mechanism causing selective metabolic inhibition in target cells. J Immunol 138: 550–565
Stuehr DJ, Nathan CF (1989) Nitric oxide. A macrophage product responsible for cytostasis and respiratory inhibition of tumor target cells. J Exp Med 169: 1543–1555
Keller R, Geiges M, Keist R (1990) l-arginine-dependent reactive nitrogen intermediates as mediators of tumor cell killing by activated macrophages. Cancer Res 50: 1421–1425
Kröncke KD, Funda J, Berschick B, Kolb H, Kolb-Bachofen V (1991) Macrophage cytotoxicity towards isolated islet cells: neither lysis nor its protection by nicotinamide are beta-cell specific. Diabetologia 34: 232–238
Appels B, Burkart V, Kantwerk-Funke G, Funda J, KolbBachofen V, Kolb H (1989) Sponteanous cytotoxicity of macrophages against pancreatic islet cells. J Immunol 142: 3803–3808
Kröncke KD, Kolb-Bachofen V, Berschick B, Burkart V, Kolb H (1991) Activated macrophages kill pancreatic syngeneic islet cells via arginine-dependent nitric oxide generation. Biochem Biophys Res Commun 175: 752–758
Wood KS, Buga GM, Byrns RE, Ignarro LJ (1990) Vascular smooth muscle-derived relaxing factor (MDRF) and its close similarity to nitric oxide. Biochem Biophys Res Commun 170: 80–88
Heinrikson RL, Meredith SC (1984) Amino acid analysis by reverse-phase high-performance liquid chromatography: precolumn derivatization with phenylisothiocyante. Anal Biochem 136: 65–74
Ding AH, Nathan CF, Stuehr DJ (1988) Release of reactive nitrogen intermediates from mouse peritoneal macrophages. J Immunol 141: 2407–2412
McCall TB, Feelisch M, Palmer RJM, Moncada S (1991) Identification of N-iminoethyl-l-ornithine as an irreversible inhibitor of nitric oxide synthase in phagocytic cells. Br J Pharmacol 102: 234–238
Lambert LE, Whitten JP, Baron BM, Cheng HC, Doherty NS, McDonald IA (1990) Nitric oxide synthesis in the CNS, endothelium and macrophages differs in its sensitivity to inhibition by arginine analogues. Life Sci 48: 69–75
Feelisch M, Noack EA (1987) Correlation between nitric oxide formation during degradation of organic nitrates and activation of guanylate cyclase. Eur J Pharmacol 139: 19–30
Leeuwenkamp OR, van Bennekom WP, van der Mark EJ, Bult A (1984) Nitroprusside, antihypertensive drug and analytical reagent; review of (photo)stability, pharmacology and analytical properties. Pharm Weekbl 6: 129–140
Lang K (1933) Die Rhodanbildung im Tierkörper. Biochem Z 259: 243–256
Aird BA, Horowitz PM (1988) The differential functional stability of various forms of bovine liver rhodanese. Biochim Biophys Acta 956: 30–38
Ignarro LJ (1992) Haem-dependent activation of cytosolic guanylate cyclase by nitric oxide: a widespread signal transduction mechanism. Biochem Soc Trans 20: 465–469
Laychock SG (1981) Evidence for guanosine 3′,5′-monophosphate as a putative mediator of insulin secretion from isolated rat islets. Endocrinology 108: 1197–1205
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: 3043–3052
Francis SH, Noblett BD, Todd BW, Weels JN, Corbin JD (1988) Relaxation of vascular and tracheal smooth muscle by cyclic nucleotide analogs that preferentially activate purified cGMP-dependent protein kinase. Mol Pharmacol 34: 506–517
Iyengar R, Stuehr DJ, Marletta MA (1987) Macrophage synthesis of nitrite, nitrate and N-nitrosamines: precursors and role of the respiratory burst. Proc Natl Acad Sci USA 84: 6369–6373
Malaisse WJ, Malaisse-Lagae F, Sener A, Pipeleers DG (1982) Determinations of the selective toxicity of alloxan to the pancreatic B cell. Proc Natl Acad Sci USA 79: 927–930
Radi R, Beckman JS, Bush KM, Freeman BA (1991) Peroxynitrite oxidation of sulfhydryls. The cytotoxic potential of superoxide and nitric oxide. J Biol Chem 266: 4244–4250
Garg UC, Hassid A (1989) Inhibition of rat mesangial cell mitogenesis by nitric oxide-enerating vasodilators. Am J Physiol 257: F60-F66
MacIntyre I, Zaidi M, Towhidul ASM et al. (1991) Osteoclastic inhibition: an action of nitric oxide not mediated by cyclic GMP. Proc Natl Acad Sci USA 88: 2936–2940
O'Connor KJ, Knowles RG, Patel KD (1991) Nitrovasodilators have proliferative as well as antiproliferative effects. J Cardiovasc Pharmacol 17 [Suppl 3]: S100-S103
Bonner-Weir S, Orci L (1982) New perspectives on the microvasculature of the islets of Langerhans in the rat. Diabetes 31: 883–889
Samols E, Stagner JI, Ewart RB, Marks V (1988) The order of islet microvascular cellular perfusion is B→A→D in the perfused rat pancreas. J Clin Invest 82: 350–353
Southern C, Schulster D, Green IC (1990) Inhibition of insulin secretion by interleukin-1β and tumor necrosis factor-α via an l-arginine-dependent nitric oxide generating mechanism. FEBS Lett 276: 42–44
Welsh N, Eizirik DL, Bendtzen K, Sandler S (1991) Interleukin1β-induced nitric oxide production in isolated rat pancreatic islets requires gene transcription and may lead to inhibition of the Krebs cycle enzyme aconitase. Endocrinology 129: 3167–3173
Corbett JA, Lancaster JR Jr, Sweetland MA, McDaniel ML (1991) Interleukin-1β-induced formation of EPR-detectable iron-nitrosyl complexes in islets of Langerhans. J Biol Chem 266: 21351–21354
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. Biochem Biophys Res Commun 182: 333–340
Hibbs JB, Taintor RR, Vavrin Z (1984) Iron depletion: possible cause of tumor cell cytotoxicity induced by activated macrophages. Biochem Biophys Res Commun 123: 716–723
Drapier IC, Hibbs JB (1986) Murine cytotoxic activated macrophages inhibit aconitase in tumor cells. J Clin Invest 78: 790–797
Granger DL, Lehninger AL (1982) Sites of inhibition of mitochondrial electron transport in macrophage-injured neoplastic cells. J Cell Biol 95: 527–535
Lepoivre M, Fieschi F, Coves J, Thelander L, Fontcave M (1991) Inactivation of ribonucleotide reductase by nitric oxide. Biochem Biophys Res Commun 179: 442–448
Kwon NS, Stuehr DJ, Nathan CF (1991) Inhibition of tumor cell ribonucleotide reductase by macrophage-derived nitric oxide. J ExpMed 174: 761–767
Wink DA, Kasprzak KS, Maragos CM et al. (1991) DNA deaminating ability and genotoxicity of nitric oxide and its progenitors. Science 254: 1001–1003
Nguyen T, Brunson D, Crespi CL, Penman BW, Wishnok JS, Tannenbaum SR (1992) DNA damage and mutation in human cells exposed to nitric oxide in vitro. Proc Natl Acad Sci USA 89: 3030–3034
Lukic ML, Stosic-Grujicic S, Ostojic N, Chan WL, Liew FY (1991) Inhibition of nitric oxide generation affects the induction of diabetes by streptozotocin mice. Biochem Biophys Res Commun 178: 913–920
Kolb H, Kiesel U, Kröncke KD, Kolb-Bachofen V (1991) Suppression of low dose streptozotocin induced diabetes in mice by administration of nitric oxide synthase inhibitor. Life Sci 49: PL213-PL217
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kröncke, K.D., Rodriguez, M.L., Kolb, H. et al. Cytotoxicity of activated rat macrophages against syngeneic islet cells is arginine-dependent, correlates with citrulline and nitrite concentrations and is identical to lysis by the nitric oxide donor nitroprusside. Diabetologia 36, 17–24 (1993). https://doi.org/10.1007/BF00399088
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00399088