Antiangiogenic activity of extracts and fractions from an endemic plant Ardisia pyramidalis (Cav.) Pers. From Bataan, Philippines using Duck in ovo chorioallantoic membrane assay

Using bioassay directed fractionation, the angiosuppressive activity of leaf extracts of Ardisia pyramidalis (Myrsinaceae) was determined using the duck in ovo chorioallantoic membrane (CAM) assay. The methanol extract was antiangiogenic as indicated by the statistically significant lower blood vessel count compared with the negative control and the untreated eggs. The hexane partition was observed to be more angiosuppressive compared to the ethyl acetate counterpart. Fractionation by normal phase vacuum column chromatography revealed that the angiosuppressive agents were relatively non-polar. Hexane fraction 4 was the most potent. Further fractionation into subfractions 4.4.2 - 4.4.9 showed decreasing potency which was not comparable to the positive control. Subfractionation showed loss of antiangiogenic activity which may indicate synergistic effects in the parent fraction. Immunohistochemical assays and transmission electron microscopy results support the findings of the CAM assay.