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Research Article - Asian Journal of Biomedical and Pharmaceutical Sciences (2017) Volume 7, Issue 61
In vitro Antiplasmodial Activity and Cytotoxicity of Extracts and Fractions of Bidens pilosa
Malaria is one of the most important health problems in Cameroon. The aim of this study was to evaluate the in vitro activity of the extracts and fractions of Bidens pilosa on Plasmodium falciparum strains. Plasmodium falciparum were cultivated by the method of Trager and Jensen (1976) and maintained in fresh O+ human erythrocytes at 4 % haematocrit in complete medium (RPMI 1640 with Albumax II). The ring stage synchronized P. falciparum strains Pf3D7 (Chloroquine sensitive) and Pf INDO (Chloroquine resistant), 1% parasitemia, 2% hematocrit} were incubated in a 96-well microplate for 48 h with different concentrations of plant extracts and fractions of the most active extract ranging from 1.56 to 100 µg/ml. RPMI and 0.4% DMSO were used as negative controls; while Chloroquine (1 µM) was used as positive control. The results were obtained by the microtiter plate using SYBR Green I fluorescence assay. Cytotoxicity was determined against HEPG2 and L929 cells using MTT assay. The most potent extract was chromatographed on reverse phase HPLC towards antiplasmodial activity guided purification of metabolites. Of the 3 plant extracts tested, the highest antiplasmodial (IC50 of 8.18 ±2.15 µg/ml) activity was observed with Ethyl Acetate extract of the leaves of Bidens pilosa. This extract was then selected for antiplasmodial activity guided fractionation and isolation of active fractions. Neighbouring fractions were combined judiciously to ensure maximum purity to obtain 15 pools. Among the 15 pools evaluated, the most potent was number 12 with an IC50 of 0.73 ± 0.07 µg/ml (Pf 3D7) and 3.53 ± 1.85 µg/ml (Pf INDO). Pool 12 exhibited a CC50 of 88.34 ± 1.56 µg/ml (HEPG2 cells) and 81.31 ± 2.42 µg/ml (L929 cells). Therefore, pool 12 can be considered as highly active and non-toxic. Our findings, therefore, corroborate the use of B. pilosa as antimalarial in Cameroonian folk medicine.
Author(s): Noumedem Anangmo Christelle Nadia, Wabo Pone J, Kumar Kaushi N, Mohana krishnan D, Mittal Garima, Nguemfo Tchakugni Arlette, Mishral Shubbi, Mpoame Mbida, Dinkar Sahal