Anti angiogenic effects of hydroalcoholic fenugreek seed extract on huvecs

Angiogenesis involves the differentiation, proliferation and migration of endothelial cells, leading to tubulogenesis and the formation of vessels. This process is essential in physiological and phathologicalconditions such as tumor growth. The significance of angiogenesis in tumourigenesis has been well demonstrated, leading to anti-angiogenesis as an important strategy in cancer therapy and developmentof the effective drugs with the less side effects. Synthetic medicines cause serious side effects and are associated with some forms of drug resistances. Therefore utilization of natural compounds includingmedicinal plants could be an alternative concept to inhibit angiogenesis and prevent the development of tumors. The fenugreek is a natural compound that has been used in the different Researches forinhibition of angiogenesis. In this study,in order to assess the anti-angiogenic effects of fenugreek seed extract on the human umbilical vein endothelial cells (HUVECs), we evaluated the Expression levels ofgrowth factors such as VEGF, bFGF and matrix metaloproteinases such as MMP-2 and MMP-9 plus activity of the enzymes(Zymography). Fenugreek extract IC50 value was determined using MTT assay.HUVECs were treated with different doses of Fenugreek extract. Expression levels of MMP-2, MMP- 9, VEGF and bFGF was explored by Real-time PCR. Also activity of MMPs was measured by gel zymography. The amount of IC50 for HUVECs was 100 μg/ml in 24h time. Real time PCR results indicate that MMP2 gene expression in treated cells with concentration 50 μg/ml, 100μg/ml and 200μg/ml of extract, in comparison of control group, decreased to 0.6, 0.3 and 0.3, respectively (p<0.001). Also MMP9 gene expression in treated cells with concentration 50 μg/ml, 100μg/ml and 200 μg/ml ofextract, in comparison of control group decreased to 0.7 (p< 0.01), 0.3 and 0.1 (p<0.001), respectively.For growth factors, Gene expression for bFGF at concentration 100μg/ml and 200 μg/ml of extract, observed significant decreased to 0.3 and 0.2 in comparison of control group (p< 0.05). Also VEGF gene expression had a reverse relation with dose increasing and at concentrations 50 μg/ml,100μg/ml and 200 μg/ml of extract, in comparison of control group decreased to 0.7, 0.3 (p< 0.05) and 0.2 (p<0.01), respectively. Analysis of zymogram for MMP2 showed that enzyme activity atconcentrations 100 μg/ml and 200 μg/ml of extract, decreased to 70.42% and 56.5% in comparison of control group(p< 0.001). Also MMP9 activity at concentrations 50 μg/ml, 100μg/ml and 200 μg/ml ofextract, in comparison of control group decreased to 77.3%(p< 0.01), 68.6% and 66.1% (p<0.001), respectively. The effect of Fenugreek seed extract on Expression and activity of Matrix Metaloproteinases and expression of growth factors was studied for the first time which showed that Fenugreek extract can inhibit angiogenesis through reduction of VEGF and bFGF expression and following it the reduction of MMP-2 and MMP-9 expression and Activity of them.

کلیدواژه ها:

Cell and Cancer ، Solid Tumors ، Cancer Treatment and Management ، Targeted Cancer Therapy ، Drugs and Cancer