Abstract

β-Carotene (β-Crt) can be dispersed in hydrophobic regions of the membrane of red blood cells (RBC). Its location, orientation and distribution strongly depend on carotenoid concentration. In the present pilot trial (six human subjects involved), it is demonstrated that incubation of RBCs with β-Crt (1.8 × 107 β-Crt molecules per RBC, 50 μmol/L) results in expansion of the membrane of RBCs and slight elongation of the cell. The changes are of statistical significance, as verified by the Wilcoxon test at p < 0.05. They indicate (i) a highly random orientation and location of β-Crt inside the membrane and (ii) a tendency for its interaction with membrane skeleton proteins. The accompanying effect of decreased RBC resistance to lysis is possibly a result of the incorrect functioning of ion channels due to their modification/disruption. At higher β-Crt concentrations, its clustering inside membranes may occur, leading to further alterations in the shape and size of RBCs, with the most pronounced changes observed at 1.8 × 108 β-Crt molecules per RBC (500 μmol/L). Due to the reduced permeability of ions, such membranes exhibit increased resistance to haemolysis. Finally, we show that interactions of β-Crt with the membrane of RBCs lead to an alteration in haemoglobin-oxygen affinity, shifting the oxyhaemoglobin dissociation curve toward higher oxygen partial pressures. If the impact of β-Crt on a curve course is confirmed in vivo, one may consider its role in the fine tuning of O2 transportation to tissues. Hence, at low concentrations, providing unchanged elastic and functional properties of RBCs, it could serve as a beneficial agent in optimising heart performance and cardiovascular load.

Highlights

  • Carotenoids are a ubiquitous group of natural pigments

  • The suspension of red blood cells (RBC) was divided into 5 equal fractions containing: (i) control sample and (ii–v) erythrocytes treated with different β-Crt concentrations (25 μmol/L, 50 μmol/L, 100 μmol/L and 500 μmol/L, respectively)

  • To follow the effect of β-Crt on the reversibility of O2 binding to Hb in RBCs, Mössbauer spectroscopy was applied according to the described procedure [13]

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Summary

Introduction

Carotenoids are a ubiquitous group of natural pigments In biological systems, they fulfil several important functions, among which their antioxidant activity is regarded as the most significant. Its main functions (transport of O2 and CO2 , buffering of H+ ions, NO metabolism) are already well-established It is an allosteric protein: binding of one O2 to one heme-iron (HFe) increases the O2 affinity within the remaining groups. The Mössbauer data, along with the osmotic fragility assay, pointed to an unexpected, distinctive effect of β-Crt as a modulator of (i) O2 -binding properties of Hb and (ii) the membrane features of erythrocytes, such as their stability and permeability to ions, extending our knowledge in this field of science. The direction of changes is discussed in view of the potential role of β-Crt under hypoxia conditions [21]

Sample Preparation
Optical Imaging of RBCs
Osmotic Fragility Test
Mössbauer Spectroscopy
Morphometric
O2 partial presgests that Hb in 500
Comparison inin
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