Than42抑制血管內皮生長因子引發之血管新生作用機轉之研究

Abstract

Angiogenesis is a multistep process of new blood vessel formation from preexisting vasculature, which is essential in many pathological disorders. Endothelial cells play a critical role in the processes of angiogenesis including cell proliferation, migration, differentiation and tube formation. Anti-angiogenesis is a therapeutic strategy for certain cancers. In the present study, we found Than42, 5-(4-Hydroxy-3-methoxyphenyl)-3-(5-methyl-2-furyl)-1-phenylpyrazole inhibited human umbilical vein endothelial cells (HUVECs) growth under vascular endothelial growth factor (VEGF) stimulation by MTT asssay. Than42 preferentially inhibited the adhesion of HUVECs to fibrinogen by affecting B3 integrin affinity/expression. In addition, Than42 inhibited VEGF-induced tube formation and cell migration in vitro in a concentration-dependent manner. By using Annexin V/PI double staining, we found Than42 inhibited cell growth by inducing apoptosis. Than42 also concentration-dependently blocked VEGF-induced reactive oxygen species (ROS) production. In regard to intracellular signal transduction, Than42 blocked the activation of PI3K/AKT, ERK1/2, Rac1/Cdc42, endothelial NO synthase (eNOS) and the nuclear translocation of NF-kB stimulated by VEGF. VEGF-induced matrix metalloproteinase (MMP)-2 protein and mRNA expression were also decreased by the treatment of Than42. In addition, VEGF-induced FAK phosphorylation and actin cytoskeleton reorganization in HUVECs were affected by Than42. Under hypoxia condition, Than42 also concentration-dependently interfered with the activation of HIF-1d and Akt. Besides, Than42 inhibited VEGF-induced angiogenesis in Matrigel plug implantation assay in vivo. These results indicate that Than42 exhibits anti-angiogenic activity both in vivo and in vitro through the blockade of NF-kB and VEGF-VEGFR-2 signaling pathways, suggesting that Than42 could be a potential compound as an anti-angiogenic agent.