Mechanism of hemolysis of human erythrocytes exposed to monosodium urate monohydrate crystals. Preliminary characterization of membrane pores

Abstract

Microcrystals of monosodium urate monohydrate (MSUM) have the ability to cause rapid hemolysis of erythrocytes. The nature of the initial MSUM crystal-erythrocyte membrane binding interaction was investigated over a range of different ionic strength media. There was negligible binding of MSUM to erythrocyte ghost membranes in low ionic strength media such as isotonic mannitol but binding was dramatically increased in isotonic NaCl/mannitol solutions or isotonic mannitol containing 1 mM Ca2+. Hemolysis induced by MSUM crystals was preceded by the leakage of K+ from the cells suggesting a colloid-osmotic mechanism of hemolysis. The inclusion of large (oligosaccharide) molecules in the extracellular media or the modulation of the extracellular solution tonicity inhibited both the rate and extent of hemolysis supporting the concept of MSUM-induced pores followed by colloid osmotic hemolysis.

Genistein abrogates pre-hemolytic and oxidative stress damage induced by 2,2′-Azobis (Amidinopropane)

“…Hemolysis can occur either by direct membrane disruption (Silva et al, 2000) or by pore formation (Jackson et al, 1996). In the latter case, a rapid efflux of K + precedes a slower release of hemoglobin, and both processes show a biphasic character (Jackson et al, 1996).…”

Hydroxytamoxifen interaction with human erythrocyte membrane and induction of permeabilization and subsequent hemolysis

“…induced hemolysis occurs via an osmotic swelling. In fact, the osmotic hemolysis described for several compounds (Lieber et al, 1984;Deuticke et al, 1989;Jackson et al, 1996) involves the formation of permeability paths for small ions, which allow water osmosis as consequence of the increased osmotic pressure inside the erythrocyte. The osmotic imbalance causes cells to swell, inducing physical rupture of the membrane followed by hemoglobin leakage.…”

“…The osmotic fragility experiments were carried out according to standardized procedures (Takeuchi et al, 1989;Jackson et al, 1996) in phosphate buffered solutions (10 mm sodium phosphate, pH 7.4) with increasing concentrations of NaCl and a hematocrit of 0.33%. After centrifugation at 3000 rpm for 10 min, the absorbance of supernatants was read at 540 nm.…”

“…To further explore this permeabilizing effect, the efflux of intracellular K + in parallel with hemoglobin release was searched, since K + efflux has revealed a reliable methodology for the assay of permeability changes induced by several hemolytic compounds (Cybulska et al, 1995;Jackson et al, 1996). Fig.…”

Pycnogenol® prevents haemolytic injury in G6PD deficient human erythrocytes

“…The efflux of K + is considered as a reliable indicator of an altered erythrocyte membrane permeability (Cybulska et al, 1995;Jackson et al, 1996) which typically precedes haemoglobin release from the haemolysing cell (Cruz-Silva et al, 2002). Since quinine is a blocker (not activator) of K + channels (Grinstein and Foskett, 1990;Kuriyama et al, 1995;Shimizu et al, 2001) its most likely mechanism for haemolytic action in G6PD deficient erythrocytes appears to be a cell damage caused by the formation of free radicals.…”