Abstract
Iron is needed for life-essential processes, but free iron overload causes dangerous clinical consequences. The study of the role of red blood cells (RBCs) in the influence of excess free iron in the blood on the pathological consequences in an organism is relevant. Here, in a direct biophysical experiment in vitro, we studied the action of free iron overload on the packed red blood cell (pRBC) characteristics. In experiments, we incubated pRBCs with the ferrous sulfate solution (Fe2+). Wе used free iron in a wide range of concentrations. High Fe2+ concentrations made us possible to establish the pattern of the toxic effect of excess iron on pRBCs during a reduced incubation time in a biophysical experiment in vitro. It was found that excess free iron causes changes in pRBC morphology, the appearance of bridges between cells, and the formation of clots, increasing the membrane stiffness and methemoglobin concentration. We created a kinetic model of changes in the hemoglobin derivatives. The complex of simultaneous distortions of pRBCs established in our experiments can be taken into account when studying the mechanism of the toxic influence of excess free iron in the blood on pathological changes in an organism.
Highlights
Overload of free iron in the blood has different origins: chronic iron overload is a common complication of repeated packed red blood cell transfusions [1, 4,5,6]; disorders of iron metabolism and chronic iron overload are observed in a number of acute and chronic inflammatory diseases and in acute respiratory distress syndrome [7, 8]; cell-free hemoglobin can become a source of excess free iron, arising under the action of hemolytic poisons [9], sickle cell anemia [10], hemodialysis [11], cardiac bypass [12], pulmonary arterial hypertension [13, 14], and sepsis [15,16,17,18], with phagocytosis of damaged red blood cells (RBCs) [19, 20], hereditary hemochromatosis [21], and viral diseases [22,23,24,25], including SARS-CoV-2 [8, 26, 27]
KG, Germany). is procedure was carried out twice. en, the lysate (L) was prepared by mixing 15 μl of packed red blood cell (pRBC) and 100 μl of distilled water, and the pRBC suspension (S) was prepared by mixing 15 μl of pRBCs and 100 μl of phosphate buffer saline (PBS). en, 100 μl of lysate/ suspension were diluted in 4.1 ml of solution B. e time of iron interaction with pRBCs was named as the incubation time and designated as tinc
100 μl of lysate was diluted in 4 ml of distilled water, or 100 μl of suspension was diluted in 4 ml of PBS with 100 μl solution A as shown in Figures 1(b) and 1(c). e working solution with ferrous sulfate and lysate was designated as LFe2+ 1700, and the similar suspension solution was designated as SFe2+1700; the solution without ferrous sulfate for lysate was designated as LFe2+ 0, and the suspension was designated as SFe2+ 0. e incubation times of the lysate of pRBCs were 0, 15, 30, 45, 60, 90, 120 min, and 24 hours, and for the suspension of pRBCs, the incubation times were 1, 5, and 24 hours
Summary
Iron is a significant metal for various processes in biological systems, but there is no active mechanism for the excretion of excess iron from the body under conditions of iron overload (hemochromatosis) [1,2,3]. Pathological consequences are associated with the action of various factors on elements and processes in the blood in the cardiovascular system: the coagulation system is changed, blood clotting, hemostasis, and thrombosis that is clinically significant are arisen [31, 40, 41]. In these cases, RBCs are the key elements. High Fe2+ concentrations made us possible to establish the pattern of the toxic effect of excess iron on pRBCs during a reduced incubation time in a biophysical experiment in vitro. We created a mathematical kinetic model of changes in the concentrations of Fe2+ and hemoglobin derivative concentrations, which adequately describes the experimental data and allows to estimate the rate constants of redox processes
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