Amino Acids Extract from Black Bean Seeds (Phaseolus Vulgaris L.)Inhibit Hemoglobin Polymerizationand Modulate Erythrocyte Membrane Atpases In Sickle Cell Disease (SCD)

Abstract

Back ground: The basic extracts involved in the formulation of functional foods/nutraceutics must first be the subject of preclinical and clinical trials. The several studies were shown that amino acids extract from black bean seeds (Phaseolus vulgaris L.) have antisickling, antioxidant and anti-haemolitic properties. Objective: The objective of this study is to investigate the effect of amino acids extract from black bean seeds (Phaseolus vulgaris L.) on hemoglobin polymerization and erythrocytes membrane ATPases activities in Sickle Cell Disease (SCD). Method: The ability for the amino acids extract to inhibit sickle cell hemoglobin polymerization and improve the Fe2+/Fe3+ ratio of HbSS blood were assed using spectroscopic method. The activities of three ATPases (Na+/K+; Ca2+; and Mg2+-ATPases) were investigated by the kinetic method following the release of inorganic phosphate (Pi) by ATP taking into account the determination of the total membrane proteins of erythrocytes by Bradford’s method. Result: The lowest potential of inhibition was obtained (39.98±2%) with the amino acids extract at 12 mg/mL corresponding significantly (p<0.05) to the highest potential of inhibition of polymerization (71.16±2%). The greatest Fe2+Fe3+ ratio was obtained (3.94±2%) with the amino acids extract at 12 mg/mL corresponding significantly (p<0.05) to the best rates increase of oxy-hemoglobin (48.73±2%). The red blood cells membranes ATPase activity were estimated and expressed as μmol of Pi/mg of protein/hour x 10-3 there was an increase in the activities of Na+/ K+ and Ca2+-ATPases (255.98±62 and 282.98±36 respectively) at the concentration of 12 mg/mL of extract compared to the negative control (167.36±60 and 236.91±62 respectively). However, for Mg2+-ATPase, there is rather a significant reduction (p<0.05) in this activity (286.71±51 against 146.53±35) obtained in the presence of the amino acid extract at 12 mg/mL). Conclusion: The amino acids extract of black bean seeds (Phaseolus vulgaris L.) at a concentration of 12 mg/mL offered the best inhibition potentials of hemoglobin S polymerization, increase rates of oxyhemoglobin and the best ATPasic activities. Keywords: Amino acid extracts; black bean seeds; sickle cell disease; polymerization; Fe2+/Fe3 + ratio; ATPase activities