IN VITRO ANTI-SICKLING EFFECT AND ACTIVITY GUIDED FRACTIONATION OF AGERATUM CONYZOIDES AND MANGIFERA INDICA METHANOLIC EXTRACTS ON HUMAN SICKLE CELLS
IN VITRO ANTI-SICKLING EFFECT AND ACTIVITY GUIDED FRACTIONATION OF AGERATUM CONYZOIDES AND MANGIFERA INDICA METHANOLIC EXTRACTS ON HUMAN SICKLE CELLS
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Date
2017-03
Authors
IBRAHIM, Danja Murja
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Abstract
All over Africa, traditional healers use medicinal plants to prepare medicines to treat a wide range of illnesses. One of these illnesses is sickle cell anemia. Sickle-cell anemia is caused as a result of substitution of glutamic acid by valine at the sixth position of beta-globin chain of hemoglobin.The effect of Ageratum conyzoides and Mangifera indica on human sickle red blood cells in vitro were investigated. The effect of the crude methanolic extracts on sickle cell was determined by sodium metabisulphite induced sickling which is based on deoxygenation of the red blood cells. The ability of the plant extract to protect the red blood cells against induced hemolysis and methemoglobin concentration were also investigated. Fractionation of the most active extract (stem bark of A. conyzoides) using column chromatography revealed eight fractions while M. indica stem bark gave seven fractions. Preliminary phytochemical screening on crude extracts of the leaf, stem and root of A. conyzoides and M. indica and their fractions showed the presence of some phytochemicals such as alkaloids, flavonoids, tannins, cardiac glycosides, carbohydrate, saponins and steroids. The results of Anti-sickling test showed that the percentage of unsickled cells were 56.93%, 64.60% and 67.77% for A. conyzoides stem bark extract, 37.60%, 44.20% and 55.50% for leaf extract and 39.50%, 43.30% and 51.90% for root extract at concentration of 0.1, 0.2 and 0.3mg/ml respectively. While 55.20%, 63.20% and 70.60% was observed for M. indica stem bark, 48.40%, 50.50% and 56.60% for leaf and 32.43%, 48.50% and 52.60% for root at 0.1, 0.2 and 0.3mg/ml respectively. Analysis of the fractions obtained from column chromatography of crude methanolic extracts shows that 70% reversal of sickling was obtained in fraction F7 of A.conyzoides, while fraction M4 of M.indica gave 74% reversal of sickling. Evaluation of the extracts for membrane protection also showed that at 0.5% sodium chloride concentration A. conyzoides reduced % lysis from 81.13% of SS control to 28.83%, 24.37% and 40.10% for stem, leaf and root extracts respectively, while M. indica gave 37.73%, 37.37% and 34.90% reduction by the stem, leaf and root respectively. The stem bark of A. conyzoides and M.indica significantly (p<0.005) decreased methemoglobinemia to values of 3.25% and 3.56% when compared with the control. Atomic absorption spectroscopy revealed the presence of magnesium, calcium, iron and zinc in both plants. The findings of this study suggest that different parts of A.conyzoides and M. indica possess promising anti-sickling activity and membrane protection capability to validate the use of the plants in the management of sickle-cell anemia by traditional practioners.
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A THESIS SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY IN PARTIAL FULFILLMENT FOR THE AWARD OF M.SC BIOCHEMISTRY
Keywords
VITRO ANTI-SICKLING EFFECT,, ACTIVITY GUIDED FRACTIONATION,, AGERATUM CONYZOIDES,, MANGIFERA INDICA METHANOLIC EXTRACTS,, HUMAN SICKLE CELLS