Ellagic Acid, the Active Compound ofPhyllanthus urinaria, ExertsIn VivoAnti-Angiogenic Effect and Inhibits MMP-2 Activity

Abstract

This study aimed to assess the potential anti-angiogenic mechanism ofPhyllanthus urinaria(P. urinaria) and characterize the major compound inP. urinariathat exerts anti-angiogenic effect. The water extract ofP. urinariaand Ellagic Acid were used to evaluate the anti-angiogenic effect in chorioallantoic membrane (CAM) in chicken embryo and human vascular endothelial cells (HUVECs). The matrix metalloproteinase-2 (MMP-2) activity was determined by gelatin zymography. The mRNA expressions of MMP-2, MMP-14 and tissue inhibitor of metalloproteinase-2 (TIMP-2) were analyzed by reverse transcription polymerase chain reaction (RT-PCR). Level of MMP-2 proteins in conditioned medium or cytosol was determined by western blot analysis. We confirmed thatP. urinaria's in vivoanti-angiogenic effect was associated with a reduction in MMP-2 activity. Ellagic acid, one of the major polyphenolic components as identified inP. urinariaby high performance liquid chromatography mass spectrometry (HPLC/MS), exhibited the same anti-angiogenic effectin vivo. BothP. urinariaand Ellagic Acid inhibited MMP-2 activity in HUVECs with unchanged mRNA level. The mRNA expression levels of MMP-14 and TIMP-2 were not altered either. Results from comparing the change of MMP-2 protein levels in conditioned medium and cytosol of HUVECs after theP. urinariaor Ellagic Acid treatment revealed an inhibitory effect on the secretion of MMP-2 protein. This study concluded that Ellagic Acid is the active compound inP. urinariato exhibit anti-angiogenic activity and to inhibit the secretion of MMP-2 protein from HUVECs.