A NOVEL IN-VITRO REVERSAL OF SICKLED ERYTHROCYTES BY COCOS NUCIFERA WATER AND HIGH K+ -ISOTONIC SOLUTIONS.
(Abstract release date: 05/19/16)
EHA Library. Ajayi O. 06/09/16; 135017; PB2117
Dr. Olutayo Ifedayo Ajayi
Contributions
Contributions
Abstract
Abstract: PB2117
Type: Publication Only
Background
Red cell sickling and adhesion are favoured by cellular dehydration, which increases the rate of hemoglobin polymerization and cell sickling. Potassium chloride co-transport and calcium-activated potassium channel (Gardos channel) mediate erythrocyte dehydration in sickle cell disease and β-thalassemia, but their role in vaso-modulation is less defined. Blocking the potasium leakage has been suggested as a means of inhibiting dehydration and enhancing normal shape of the sickle cells. We investigated the in-vitro effect of various concentration of K+ ion in physiological solutions (PSS) as well as in cocos nucifera water which is a natural drink known for its natural high potassium content and health benefits.
Aims
This study has therefore been designed to highlight the possibility of reversing the Na+/K+ membrane counter-movement in the extra cellular medium and observe the impact on sickled erythrocyte especially in the dehydrated state using a natural medium with very high potassium but isotonic solution - Cocos nucifera water
Methods
Twenty blood samples from sickle cell anaemia subjects were collected and studied with different high potassium isotonic media. Firstly with Cocos nucifera water (with K+ of 240mM); erythrocytes sickling test was done, then repeated after incubation with the fluid for 30 mins. Another part was treated with Sodium Metabisulphite (Na2S5O7) solution to induce maximum sickling as controls, then later incubated with the fluid. Subsequently, we subjected the samples to different high concentrations of K+ in Physiological Salt Solution (PSS) (40mM, 80mM, respectively) and their percentage sickling activities were enumerated using a thin blood smear stained with Leishman dye before and after the treatment. Later experiments to study the effects of the Cocos nucifera water on Osmotic fragility of red cells from different haemoglobin protein electrophoresis before and after incubation with coconut water.
Results
The result showwed statistically significant counts of discoid shaped cells in the blood samples incubated with 80mM and Cocos nucifera water (P<0.05, respectively) over the native and the sickled- induced samples while the 40mM PSS did not show significant discoid cells over the sickled cells (P>0.05). In 80mM K+PSS, values of (18% against 78%; before and after treatment); Cocos nucifera (22% against 85%; before and after treatment) were obtained. In addition, all the samples treated with cocos nucifera water have significantly reduced osmotic fragilities (P<0.05, respectively) irrespective of their haemoglobin protein status.
Conclusion
High extracellular potassium in an isotonic medium could reverse the membrane permeability to K+ exflux from the sickled erythrocyte by possibly overwhelming the Na+/K+ counter movement across the membrane thereby promoting rehydration through K-Cl cotransporter and ultimately reverses the sickling. The inhibition of Gardos channel is suggested as a possible mechanism in the interim, also cocos nucifera water has shown in this study, the tendency to reduce osmotic pressure lysis of sickle cells and possibly prolong its life span.
Session topic: E-poster
Keyword(s): Sickle cell
Type: Publication Only
Background
Red cell sickling and adhesion are favoured by cellular dehydration, which increases the rate of hemoglobin polymerization and cell sickling. Potassium chloride co-transport and calcium-activated potassium channel (Gardos channel) mediate erythrocyte dehydration in sickle cell disease and β-thalassemia, but their role in vaso-modulation is less defined. Blocking the potasium leakage has been suggested as a means of inhibiting dehydration and enhancing normal shape of the sickle cells. We investigated the in-vitro effect of various concentration of K+ ion in physiological solutions (PSS) as well as in cocos nucifera water which is a natural drink known for its natural high potassium content and health benefits.
Aims
This study has therefore been designed to highlight the possibility of reversing the Na+/K+ membrane counter-movement in the extra cellular medium and observe the impact on sickled erythrocyte especially in the dehydrated state using a natural medium with very high potassium but isotonic solution - Cocos nucifera water
Methods
Twenty blood samples from sickle cell anaemia subjects were collected and studied with different high potassium isotonic media. Firstly with Cocos nucifera water (with K+ of 240mM); erythrocytes sickling test was done, then repeated after incubation with the fluid for 30 mins. Another part was treated with Sodium Metabisulphite (Na2S5O7) solution to induce maximum sickling as controls, then later incubated with the fluid. Subsequently, we subjected the samples to different high concentrations of K+ in Physiological Salt Solution (PSS) (40mM, 80mM, respectively) and their percentage sickling activities were enumerated using a thin blood smear stained with Leishman dye before and after the treatment. Later experiments to study the effects of the Cocos nucifera water on Osmotic fragility of red cells from different haemoglobin protein electrophoresis before and after incubation with coconut water.
Results
The result showwed statistically significant counts of discoid shaped cells in the blood samples incubated with 80mM and Cocos nucifera water (P<0.05, respectively) over the native and the sickled- induced samples while the 40mM PSS did not show significant discoid cells over the sickled cells (P>0.05). In 80mM K+PSS, values of (18% against 78%; before and after treatment); Cocos nucifera (22% against 85%; before and after treatment) were obtained. In addition, all the samples treated with cocos nucifera water have significantly reduced osmotic fragilities (P<0.05, respectively) irrespective of their haemoglobin protein status.
Conclusion
High extracellular potassium in an isotonic medium could reverse the membrane permeability to K+ exflux from the sickled erythrocyte by possibly overwhelming the Na+/K+ counter movement across the membrane thereby promoting rehydration through K-Cl cotransporter and ultimately reverses the sickling. The inhibition of Gardos channel is suggested as a possible mechanism in the interim, also cocos nucifera water has shown in this study, the tendency to reduce osmotic pressure lysis of sickle cells and possibly prolong its life span.
Session topic: E-poster
Keyword(s): Sickle cell
Abstract: PB2117
Type: Publication Only
Background
Red cell sickling and adhesion are favoured by cellular dehydration, which increases the rate of hemoglobin polymerization and cell sickling. Potassium chloride co-transport and calcium-activated potassium channel (Gardos channel) mediate erythrocyte dehydration in sickle cell disease and β-thalassemia, but their role in vaso-modulation is less defined. Blocking the potasium leakage has been suggested as a means of inhibiting dehydration and enhancing normal shape of the sickle cells. We investigated the in-vitro effect of various concentration of K+ ion in physiological solutions (PSS) as well as in cocos nucifera water which is a natural drink known for its natural high potassium content and health benefits.
Aims
This study has therefore been designed to highlight the possibility of reversing the Na+/K+ membrane counter-movement in the extra cellular medium and observe the impact on sickled erythrocyte especially in the dehydrated state using a natural medium with very high potassium but isotonic solution - Cocos nucifera water
Methods
Twenty blood samples from sickle cell anaemia subjects were collected and studied with different high potassium isotonic media. Firstly with Cocos nucifera water (with K+ of 240mM); erythrocytes sickling test was done, then repeated after incubation with the fluid for 30 mins. Another part was treated with Sodium Metabisulphite (Na2S5O7) solution to induce maximum sickling as controls, then later incubated with the fluid. Subsequently, we subjected the samples to different high concentrations of K+ in Physiological Salt Solution (PSS) (40mM, 80mM, respectively) and their percentage sickling activities were enumerated using a thin blood smear stained with Leishman dye before and after the treatment. Later experiments to study the effects of the Cocos nucifera water on Osmotic fragility of red cells from different haemoglobin protein electrophoresis before and after incubation with coconut water.
Results
The result showwed statistically significant counts of discoid shaped cells in the blood samples incubated with 80mM and Cocos nucifera water (P<0.05, respectively) over the native and the sickled- induced samples while the 40mM PSS did not show significant discoid cells over the sickled cells (P>0.05). In 80mM K+PSS, values of (18% against 78%; before and after treatment); Cocos nucifera (22% against 85%; before and after treatment) were obtained. In addition, all the samples treated with cocos nucifera water have significantly reduced osmotic fragilities (P<0.05, respectively) irrespective of their haemoglobin protein status.
Conclusion
High extracellular potassium in an isotonic medium could reverse the membrane permeability to K+ exflux from the sickled erythrocyte by possibly overwhelming the Na+/K+ counter movement across the membrane thereby promoting rehydration through K-Cl cotransporter and ultimately reverses the sickling. The inhibition of Gardos channel is suggested as a possible mechanism in the interim, also cocos nucifera water has shown in this study, the tendency to reduce osmotic pressure lysis of sickle cells and possibly prolong its life span.
Session topic: E-poster
Keyword(s): Sickle cell
Type: Publication Only
Background
Red cell sickling and adhesion are favoured by cellular dehydration, which increases the rate of hemoglobin polymerization and cell sickling. Potassium chloride co-transport and calcium-activated potassium channel (Gardos channel) mediate erythrocyte dehydration in sickle cell disease and β-thalassemia, but their role in vaso-modulation is less defined. Blocking the potasium leakage has been suggested as a means of inhibiting dehydration and enhancing normal shape of the sickle cells. We investigated the in-vitro effect of various concentration of K+ ion in physiological solutions (PSS) as well as in cocos nucifera water which is a natural drink known for its natural high potassium content and health benefits.
Aims
This study has therefore been designed to highlight the possibility of reversing the Na+/K+ membrane counter-movement in the extra cellular medium and observe the impact on sickled erythrocyte especially in the dehydrated state using a natural medium with very high potassium but isotonic solution - Cocos nucifera water
Methods
Twenty blood samples from sickle cell anaemia subjects were collected and studied with different high potassium isotonic media. Firstly with Cocos nucifera water (with K+ of 240mM); erythrocytes sickling test was done, then repeated after incubation with the fluid for 30 mins. Another part was treated with Sodium Metabisulphite (Na2S5O7) solution to induce maximum sickling as controls, then later incubated with the fluid. Subsequently, we subjected the samples to different high concentrations of K+ in Physiological Salt Solution (PSS) (40mM, 80mM, respectively) and their percentage sickling activities were enumerated using a thin blood smear stained with Leishman dye before and after the treatment. Later experiments to study the effects of the Cocos nucifera water on Osmotic fragility of red cells from different haemoglobin protein electrophoresis before and after incubation with coconut water.
Results
The result showwed statistically significant counts of discoid shaped cells in the blood samples incubated with 80mM and Cocos nucifera water (P<0.05, respectively) over the native and the sickled- induced samples while the 40mM PSS did not show significant discoid cells over the sickled cells (P>0.05). In 80mM K+PSS, values of (18% against 78%; before and after treatment); Cocos nucifera (22% against 85%; before and after treatment) were obtained. In addition, all the samples treated with cocos nucifera water have significantly reduced osmotic fragilities (P<0.05, respectively) irrespective of their haemoglobin protein status.
Conclusion
High extracellular potassium in an isotonic medium could reverse the membrane permeability to K+ exflux from the sickled erythrocyte by possibly overwhelming the Na+/K+ counter movement across the membrane thereby promoting rehydration through K-Cl cotransporter and ultimately reverses the sickling. The inhibition of Gardos channel is suggested as a possible mechanism in the interim, also cocos nucifera water has shown in this study, the tendency to reduce osmotic pressure lysis of sickle cells and possibly prolong its life span.
Session topic: E-poster
Keyword(s): Sickle cell
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