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.Keywords: Ficus deltoidea, Antiangiogenesis, Ursolic acid, Cytotoxicity, Antioxidant
Highlights
Medicinal plants have been used for centuries in folk medicine in the treatment of various diseases and today, they still play an important role in health
Phytochemical analysis of Ficus deltoidea (FD) extracts reveals the presence of relatively high content of ursolic acid, total phenolics and flavonoids with the highest concentration obtained in FD-M extract
In order to test if the antiangiogenic activity observed is due to direct cytotoxic effect on endothelial cells or due to inhibition of other angiogenesis cascade mechanisms, the cytotoxicity of FD extracts was tested against Human umbilical vein endothelial cells (HUVECs) and two cancer cell lines (HCT 116 and MDA-MB-231)
Summary
Medicinal plants have been used for centuries in folk medicine in the treatment of various diseases and today, they still play an important role in health. Ficus deltoidea (FD) Jack (Moraceae) is a traditional medicinal plant widely distributed in Southeast Asia. The plant has been utilised in treating gout, high blood pressure, pneumonia, diarrhoea, skin infections, diabetes and to improve blood circulation [3]. Previous studies have shown that FD leaves possess antinociceptive, wound healing and antioxidant properties [1,3,4]. Previous research on FD showed the presence of relatively high content of polyphenols, flavonoids, which are known as antioxidant compounds, amino acids, vitamins, carbohydrates, and purine alkaloids. Presence of a number of antioxidant compounds is known to possess various pharmacological activities including antiangiogenic and cytotoxic effects on a number of human cancer cell lines [1,2,3]
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