Herein, we report extraction of biologically important isolates from medicinally valuable plant Wedelia trilobata. The biological activities viz., anti-proliferative activity and cytotoxic effect of the methanolic extract of Wedelia trilobata was investigated by Thymidine uptake assay, wound healing assay and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, respectively. In addition, apoptotic activity was also examined through nuclear staining and DNA fragmentation assays. The IC 50 value of the methanolic extract for the growth inhibition of MEG-01 cells was found to be 80 μg/ml under which the normal HEK-293 cells were resistant to the toxic effect. Concentration and time dependent studies in Thymidine uptake and wound healing assays revealed the prominent antiproliferative activity against MEG-01 at the concentration of 80 μg/ml for 48 h. The nuclear staining and DNA fragmentation assays also confirmed that the methanolic extract induces apoptosis in MEG-01 cells.
Abstract:Background: Wedelia trilobata (L.) Hitch (WT), commonly known as yellow dots or creeping daisy, is a shrub possessing potent biological activities, and is traditionally used a medicinal plant in Ayurveda, Siddha and Unani systems of medicines, and it has also been tried against leukemia cell line MEG-01. In the present study, purification and screening of the plant was done for bioactive compounds in methanolic extract of WT for apoptotic and antileukemia activity.
Materials and methods:The methanolic extract of WT was initially purified through thin layer chromatography (TLC) and screened for the apoptotic and anti-leukemia activities. The positive band of TLC was subjected to silica gel column chromatography for further purification and the fractions obtained from it were screened again for antileukemia activity through thymidine uptake assay and apoptotic activity by DNA fragmentation, nuclear staining and flow cytometry assays. The fraction with positive result was subjected to HPLC for analysis of bioactive components. Results: Out of many combinations of solvents, the methanol and dichloromethane combination in the ratio 6:4 has revealed two bands in TLC, among which the second band showed positive results for apoptotic and anti-leukemic activities. Further purification of second band through silica gel chromatography gave five fractions in which the 3
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