Establishment of Secondary Iron Overloaded Mouse Model: Evaluation of Cardiac Function and Analysis According to Iron Concentration

Abstract

Periodic blood transfusion can lead to secondary iron overload in patients with hematologic and oncologic diseases. Iron overload can result in iron deposition in heart tissue, which decreases cardiac function and can ultimately lead to death due to dilated cardiomyopathy and cardiac failure. In this study, we established murine model of secondary iron overload, studied the changes in cardiac function with echocardiography, and examined the histopathologic changes. Three experimental groups of the six week-old C57/BL mice (H-2(b)) were injected intraperitoneally with 10 mg of iron dextran daily 5 days a week for 2, 4, and 6 weeks. Cumulative doses of iron for the three experimental groups were 100, 200, and 300 mg, while the control groups were injected with the same amounts of phosphate-buffered saline. We studied the cardiac function under anesthesia with echocardiography using a GE Vivid7 Dimension system. Plasma iron levels and liver iron contents were measured. The hearts and livers were harvested and stained with H&E and Perls Prussian blue for iron, and the levels of iron deposit were examined. We assessed the cardiac measurements after adjustment for weight. On echocardiography, thicknesses of the interventricular septum and posterior ventricular wall (PS) during diastole showed correlation with the amount of iron deposit (P < 0.01). End-diastolic volume showed dilatation of the left ventricle in the 300 mg group (P < 0.01). Changes in the fractional shortening were not statistically significant (P = 0.07). Plasma iron levels and liver iron contents were increased proportionally according to the amount of iron loaded. The histopathologic findings of PS and liver showed higher grade of iron deposit proportional to the cumulated iron dose. In this study, we present an animal model which helps understand the cardiac function changes in patients with secondary iron overload due to repeated blood transfusions. Our results may help characterize the pathophysiologic features of cardiomyopathy in patients with secondary iron overload, and our model may be applied to in vivo iron-chelating therapy studies.

Effect of Systemic Iron Overload and a Chelation Therapy in a Mouse Model of the Neurodegenerative Disease Hereditary Ferritinopathy

“…No major differences were observed during the pathologic examination of the organs; however, iron-containing ferritin aggregates in the spleen and liver were remarkable different in iron loaded FTL-Tg mice (Fig 4E–4N). Ferritin aggregates in the liver were observed only in the iron-loaded group and appeared morphologically different from the intracellular IBs characteristic of HF [10, 19] (Fig 4I–4N) in agreement with previous work on iron overload in mice [20, 21]. Aggregates were stained by Abs against the Lm and H chains.…”

Effects of Iron Chelators on Pulmonary Iron Overload and Oxidative Stress in β-Thalassemic Mice

“…The iron-loading procedure in BKO mice has been described previously [11,12] . Briefly, mice were given iron dextran (20 mg iron/mouse, Sigma, St. Louis, Mo., USA) via intraperitoneal in 9 doses over a 2-week period (180 mg iron/mouse total).…”

“…To mimic pulmonary iron burdens in thalassemia patients who receive repeated blood transfusions, peritoneal injections of iron dextran were used in BKO mice. Modification of the duration and dosing schedule of iron dextran allowed for fine control of the degree of iron overload [11] . We previously reported on the use of this model to examine changes in hepatic and cardiac pathology when titrating ironoverload levels to be similar to those seen in thalassemia major.…”

Hepcidin knockout mice as a model of iron-overload associated liver disease

“…On the other hand, comparable or even higher hepatic iron levels were reported in animals fed with ferrocene or receiving intraperitoneal supplementation with iron dextran or ferric ammonium citrate. However, these models are of limited value since they are achieved by non-physiological iron uptake mechanisms and do not mimic the chronic, slow increase of iron levels seen in hemochromatosis subjects [124,125,126,127,128,129].…”