Evaluation of antioxidant and antiproliferative activity of Flueggea leucopyrus Willd (katupila)

BACKGROUND Flueggea leucopyrus Willd is a shrub grown in many parts of the dry zones in Sri Lanka. The leaves of F. leucopyrus has been used for treating cancer in the traditional system of medicine in Sri Lanka. Hence, this study was performed to analyze the antioxidant and antiproliferative properties of the aqueous extract of the leaves of F. leucopyrus on HEp-2 cells. METHOD The aqueous extract of F. leucopyrus leaves (AEFLL) was freeze dried. Total phenolic content was assayed using Folin Ciocalteu reagent. Antioxidant activities of the extracts were evaluated using in vitro assays: inhibition of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging and 2-deoxy-D-ribose degradation assay. Nitric oxide radical scavenging activity was determined by using Griess reagent. The MTT, LDH assays and protein synthesis were used to study antiproliferative and cytotoxic activities against the Hep-2 cell after 24 hour exposure. DNA fragmentation and microscopic examination of cells stained with a mixture of ethidium bromide/acridine orange were used to visualize apoptosis in HEp-2 cells treated with the AEFLL. RESULTS The total phenolic content of the extract was 22.15 ± 1.65 (w/w) % of gallic acid equivalent. The values for EC50 were 11.16 ± 0.37, 4.82 ± 1.82 and 23.77 ± 3.16 μg/mL for DPPH radical scavenging, nitric oxide radical scavenging activity and 2-deoxy-D-ribose degradation assay respectively. The EC50 with MTT and LDH assays were 506.8 ± 63.16 and 254.52 ± 42.92 μg/mL respectively. A dose dependent decrease in protein synthesis in HEp-2 cells was shown with an EC50 value of 305.84 ± 12.40 μg/mL. DNA fragmentation and ethidium bromide/acridine assays showed that the AEFLL induces apoptosis in HEp-2 cells. These results were in conformity with the morphological changes observed in the cells treated with the AEFLL. The brine shrimp bioassay showed that the AEFLL had no lethality over the concentration range of 50-500 μg/mL. CONCLUSIONS Aqueous extract of the leaves of F. leucopyrus extract demonstrated antioxidant activity in vitro. Further it showed antiproliferative properties and induced apoptosis in HEp-2 cells.