Summary
An experimental model of secondary hemochromatosis is described. Saccharated iron was administered i.v. to rats for 7 months in total doses in the range 1.0–1.7 g per kg body weight. After the completion of iron loading, the biochemical measurements revealed elevation of alanine aminotransferase (ALT), slight reduction of plasma glucose concentration, and significant reduction of both plasma and liver ascorbic-acid levels. The mean liver iron concentration was 50 times higher in iron-loaded animals than in controls. High concentrations of inorganic iron were also observed in spleen, pancreas, and heart. Histologic analysis revealed marked hepatic fibrosis in most animals in the experimental group. These results demonstrate this animal model presents some pathologic findings observed in human transfusional hemochromatosis. Additionally, hydroxyl free radicals were detected by electron paramagnetic resonance (EPR) spectrscopy in the iron-overloaded liver tissue processed at pH 5.0. No free radicals were detected at pH 7.4. These results suggest the possible participation of hydroxyl free radicals in the cellular toxicity of iron overload.
Similar content being viewed by others
References
Andersson NSE (1950) Experimental and clinical investigations into the effect of parenterally administered iron. Acta Med Scand 138 (Suppl 241):1–71.
Bacon BR, Britton RS (1990) The pathology of hepatic iron overload: a free radicalmediated process? Hepatology 11:127–137
Bacon BR, Tavill AS, Brittenham GM, Park CH, Recknagel RO (1983) Hepatic lipid peroxidation in vivo in rats with chronic iron overload. J Clin Invest 71:429–439
Barry M, Sherlock S (1971) Measurement of liver-iron concentration in needle-biopsy specimens. Lancet I:100–103
Bassett ML, Halliday JW, Powell LW (1986) Value of hepatic iron measurements in early hemochromatosis and deterimination of the critical iron level associated with fibrosis. Hepatology 6:24–29
Bothwell TH, Bradlow BA (1960) Siderosis in the Bantu. Arch Pathol 70:279–292
Brissot P, Campion JP, Guillouzo A, Allain H, Messner M, Simon M, Ferrand B, Bourel M (1983) Experimental hepatic iron overload. Dig Dis Sci 28:616–624
Brown EB, Dubach R, Smith DE, Reynafarje C, Moore CV (1957) Studies in iron transportation and metabolism: X. Long-term iron overload in dogs. J Lab Clin Med 50:862–893
Brown EB, Smith DE, Dubach R, Moore CV (1959) Lethal iron overload in dogs. J Lab Clin Med 53:591–606
Cappell DF (1930) The late results of intravenous injection of colloidal iron. J Pathol Bacteriol 33:175–196
Fairbanks VF, Baldus WP (1990) Disorders of iron metabolism. In: Williams WJ, Beutler E, Erslev AJ, Lichtman MA (eds) Hematology, 4th edn. McGraw-Hill, New York, pp 752–758
Finch CA, Hegsted M, Kinney TD, Thomas ED, Rath CE, Haskins D, Finch S, Fluharty RG (1950) Iron metabolism. The pathophysiology of iron storage. Blood 5:983–1008.
Fletcher LM, Roberts FD, Irving MG, Powell LW, Halliday JW (1989) Effects of iron loading on free radical scavenging enzymes and lipid peroxidation in rat liver. Gastroenterology 97:1011–1018
Green MJ, Hill HAO (1984) Chemistry of dioxygen. Methods Enzymol 105:3–22
Golberg L, Smith JP, Martin LE (1957) The effects of intensive and prolonged administration of iron parenterally in animals. Br J Exp Pathol 38:297–311
Golberg L, Martin LE, Batchelor A (1962) Biochemical changes in the tissues of animals injected with iron: 3. Lipid peroxidation. Biochem J 83:291–298
Gutteridge JMC, Rowley DA, Griffiths E, Halliwell B (1985) Low-molecular-weight iron complexes and oxygen radical reactions in idiopathic haemochromatosis. Clin Sci 68:463–467.
Heys AD, Dormandy TL (1981) Lipid peroxidation in iron-overladed spleens. Clin Sci 60:295–301
Holland HK, Spivak JL (1989) Hemochromatosis. Med Clin North Am 73:831–845
Hunter FE, Gebicki JM, Hoffstein PE, Weinstein J, Scott A (1963) Swelling and lysis of rat liver mitochondria induced by ferrous ions. J Biol Chem 238:828–835
Iancu TC, Shiloh H (1988) Experimental iron overload. Ultrastructural studies. Ann NY Acad Sci 526:164–178
Jacobs A (1982) Disorders of iron metabolism. In: Hoffbrand AV (ed) Recent advances in haematology, 3rd edn. Churchill Livingstone Edinburgh, pp 1–24
Krumbhaar EB, Chanutin A (1922) Studies on experimental plethora in dogs and rabbits. J Exp Med 35:847–871
Lisboa PE (1971) Experimental hepatic cirrhosis in dogs caused by chronic massive iron overload Gut 12:363–368
Lowry OH, Lopez JA, Bessey OA (1945) The determination of ascorbic acid in small amounts of blood serum. J Biol Chem 160:609–615
McKnight RC, Hunter FE, Oehlert WH (1965) Mitochondrial membrane ghosts produced by lipid peroxidation induced by ferrous ion. I. Production and general morphology. J Biol Chem 240:3439–3446
Miller DM, Aust SD (1989) Studies of ascorbate-dependent, iron-catalyzed lipid peroxidation. Arch Biochem Biophys 271:113–119
Minotti G (1988) Metals and membrane lipid damage by oxy-radicals. Ann NY Acad Sci 551:34–44
Modell B, Berdoukas V (1984) The clinical approach to thalassaemia. Grune & Stratton, London
Nissim JA (1953) Experimental siderosis: a study of the distribution, delayed effects and metabolism of massive amounts of various iron preparations. J Pathol Bacteriol 66:185–204
O'Connell MJ, Ward RJ, Baum H, Peters TJ (1985) The role of iron in ferritin- and haemosiderin-mediated lipid peroxidation in liposomes. Biochem J 229:135–139
Ozaki M, Kawabata T, Awai M (1988) Iron release from haemosiderin and production of iron-catalyzed hydroxyl radicals in vitro. Biochem J 250:589–595
Park CH, Bacon BR, Brittenham GM, Tavill AS (1987) Pathology of dietary carbonyl iron overload in rats. Lab Invest 57:555–563
Pechet GS (1969) Parenteral iron overload. Organ and cell distribution in rats. Lab Invest 20:119–126
Poli G, Chiarpotto E, Albano E, Biasi F, Cecchini G, Dianzani MU (1986) Iron overload: experimental approach using rat hepatocytes in single cell suspension. Front Gastrointest Res 9:38–49
Polson C (1933) The failure of prolonged administration of iron to cause haemochromatosis. Br J Exp Pathol 14:73–76
Roe JH, Mills MB, Oesterling MJ, Damron CM (1948) The determination of diketo-1-gulonic acid, dehydro-1-ascorbic acid, and 1-ascorbic acid in the same tissue extract by the 2,4-dinitrophenylhydrazine method. J Biol Chem 174:201–208
Rous P, Oliver J (1918) Experimental hemochromatosis. J Exp Med 28:629–652
Seymor CA, Peters TJ (1978) Organelle pathology in primary and secondary haemochromatosis with special reference to lysosomal changes. Br J Haematol 40:239–253
Slott PA, Loney LA, Potter BJ (1990) The Belgrade rat as a model of hepatic transfusional iron overload in Cooley's anemia. Ann NY Acad Sci 612:565–567
Suda K (1985) Hemosiderin deposition in the pancreas. Arch Pathol Lab Med 109:996–999
Tavill AS, Bacon BR (1986) Hemochromatosis: how much iron is too much Hepatology 6:142–145
Wills ED (1969) Lipid peroxide formation in microsomes. The role of non-heme iron. Biochem J 113:325–332
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Figueiredo, M.S., Baffa, O., Neto, J.B. et al. Liver injury and generation of hydroxyl free radicals in experimental secondary hemochromatosis. Res. Exp. Med. 193, 27–37 (1993). https://doi.org/10.1007/BF02576208
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02576208