Role of membrane sialic acid and glycophorin protein in thorium induced aggregation and hemolysis of human erythrocytes

Thorium-232 (232Th), a natural radionuclide from the actinide family, is abundantly present in monazite and other ores. It is used as one of the prime fuel materials in nuclear industry and may pose an exposure risk to nuclear workers and members of the public. Human erythrocytes, as a classical cellular membrane model, were coincubated with 232Th in order to elucidate whether this naturally occurring important radionuclide produced perturbations to cell membrane. Present study revealed that erythrocytes underwent aggregation or lysis depending on the ratio of 232Th to cell. Scanning electron micrographs showed that erythrocytes transformed into equinocytes and/or spherocytes after 232Th treatment. Further examination of erythrocyte by atomic force microscopy suggested significant increase in surface roughness after 232Th treatment. Experiments on neuraminidase treated and/or anti-GpA antibody blocked erythrocytes suggested significant role of membrane sialic acid and glycophorin A (GpA) protein in aggregation or hemolytic effects of 232Th. Further results showed that 232Th caused hemolysis by colloid osmotic mechanism, as evidenced by potassium efflux, osmotic protection and osmotic fragility studies. Osmoprotection experiments indicated that hemolysis get elicited through the formation of membrane pores of ∼2.0 nm in size. Hemolysis studies in presence of inhibitors (TEA, bumetanide, DIDS and amiloride) revealed the role of K+ channel, Na+/K+/2Cl− channel, Cl−/HCO3− anion exchanger and Na+/H+ antiporter in 232Th induced erythrolysis. Presence of non-diffusible cation (N-methyl d-glucasamine) or anion (gluconate) in erythrocyte suspending medium further confirm the role of Na+ and Cl− influx in hemolytic effect of 232Th. These findings provide significant insight in structural, biochemical and osmotic toxic effects of 232Th on human erythrocytes.