Antiangiogenic effect of Ficus deltoidea jack standardised leaf extracts

Abstract

Antiangiogenic Effect of Ficus deltoidea Jack Standardised Leaf Extracts A Shafaei, NS Muslim, ZD Nassar, AFA Aisha, AMSA Majid, Z Ismail

Abstract

Purpose: To standardise the methanol and aqueous extracts of Ficus deltoidea leaf by developing a reverse phase high performance liquid chromatography (RP-HPLC) for determination of the ursolic acid content and to investigate their antiangiogenic activity. Methods: To prepare the water extract (FD-A), the powder of the plant was extracted with water under reflux for 24 h at 50 ˚C. The methanol extract (FD-M) was prepared using Soxhlet extractor for 24 h at 50 ˚C. The extracts were standardised for ursolic acid content by reverse phase high performance liquid chromatography (RP-HPLC), as well as by total phenolic and flavonoid contents. Antiangiogenic activity was studied using ex vivo rat aortic rings and in vivo chick chorioallantoic membrane angiogenesis models. The anti-proliferative effect of the extracts against normal human endothelial cells and two cancer cell lines was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Results: In rat aortic rings, methanol and water extracts inhibited the outgrowth of microvessels with IC50 values of 48.2 ± 1.1 and 62.7 ± 1.4 μg ml-1, respectively. Methanol and water extracts at doses of 100 μg disc-1 also inhibited vascularisation of chick embryo chorioallantoic membrane with inhibition values of of 62.0 ± 6.3 and 43.3 ± 4.8 %, respectively. In addition, both extracts showed potent cytotoxicity against breast and colon cancer cells while demonstrating non-cytotoxic activity against normal endothelial cells. Furthermore, the two extracts showed relatively high concentration of ursolic acid, total phenolics and flavonoids associated with potent antiangiogenic activity. Conclusion: The inhibition of angiogenesis by F. deltoidea extracts may be attributed to the relatively high ursolic acid content as well as the presence of antioxidant compounds of phenolics and flavonoids in the extracts.