Iron Deposition and Ferroptosis in the Spleen in a Murine Model of Acute Radiation Syndrome

• Published in: International journal of molecular sciences 2022-10, Vol.23 (19), p.11029
• Main Authors: Rittase, W. Bradley, Slaven, John E., Suzuki, Yuichiro J., Muir, Jeannie M., Lee, Sang-Ho, Rusnak, Milan, Brehm, Grace V., Bradfield, Dmitry T., Symes, Aviva J., Day, Regina M.
• Format: Article
• Language: English
• Subjects: Animal models; Apoptosis; Binding; Blood; Bone marrow; Captopril; Caspase-3; Cyclin-dependent kinase inhibitor p21; Denaturation; Deposition; Erythrocytes; Ferritin; Ferroptosis; Free radicals; Gene expression; Hematocrit; Hemoglobin; Immunohistochemistry; Inflammation; Investigations; Iron; Iron-binding protein; Laboratories; Leukocytes; Lysis; Macrophages; Markers; Neutrophils; Pigments; Plasma; Polarization; Post-irradiation; Protein transport; Proteins; Radiation; Radiation effects; Senescence; Spleen; Western blotting
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms231911029

Total body irradiation (TBI) can result in death associated with hematopoietic insufficiency. Although radiation causes apoptosis of white blood cells, red blood cells (RBC) undergo hemolysis due to hemoglobin denaturation. RBC lysis post-irradiation results in the release of iron into the plasma, producing a secondary toxic event. We investigated radiation-induced iron in the spleens of mice following TBI and the effects of the radiation mitigator captopril. RBC and hematocrit were reduced ~7 days (nadir ~14 days) post-TBI. Prussian blue staining revealed increased splenic Fe3+ and altered expression of iron binding and transport proteins, determined by qPCR, western blotting, and immunohistochemistry. Captopril did not affect iron deposition in the spleen or modulate iron-binding proteins. Caspase-3 was activated after ~7–14 days, indicating apoptosis had occurred. We also identified markers of iron-dependent apoptosis known as ferroptosis. The p21/Waf1 accelerated senescence marker was not upregulated. Macrophage inflammation is an effect of TBI. We investigated the effects of radiation and Fe3+ on the J774A.1 murine macrophage cell line. Radiation induced p21/Waf1 and ferritin, but not caspase-3, after ~24 h. Radiation ± iron upregulated several markers of pro-inflammatory M1 polarization; radiation with iron also upregulated a marker of anti-inflammatory M2 polarization. Our data indicate that following TBI, iron accumulates in the spleen where it regulates iron-binding proteins and triggers apoptosis and possible ferroptosis.