Identification of resveratrol derivative 3,3',4,4',5,5'-hexamethoxy- trans-stilbene as a novel pro-angiogenic small-molecule compound.

The potential to promote neovascularization in ischemic tissues using exogenous agents is an attractive avenue for therapeutics. To identify novel pro-angiogenic small-molecule compound, we screened a series of resveratrol methylated derivatives and identified 3,3',4,4', 5,5'-hexamethoxy-trans-stilbene (3,3',4,4',5,5'-HMS) potently promotes proliferation, migration, invasion and tube formation of human umbilical vein VECs (HUVECs) in vitro. Furthermore, 3,3',4,4',5,5'-HMS accelerates neo-vessels sprouting of rat aortic rings ex vivo, and neovascularization of chick chorioallantoic membrane (CAM) and mouse matrigel plugs in vivo. Microarray analyses show that the level of early growth response 1 (EGR-1), an inducible pro-angiogenic gene regulatory factor, was upregulated. The upregulation of EGR-1 was confirmed by semiquantitative RT-PCR, quantitative real-time PCR and western blotting analyses. In addition, the levels of several pro-angiogenic factors including transforming growth factor β1 (TGF-β1), vascular endothelial growth factor (VEGF), nitric oxide (NO), and the activity of endothelial NO synthase (eNOS) were elevated in 3,3',4,4',5,5'-HMS-treated HUVECs. Inhibition of NO synthase by l-NAME blocked the pro-angiogenic effects of 3,3',4,4',5,5'-HMS. Our research shows that 3,3',4,4',5,5'-HMS dramatically promoted angiogenesis in vitro, ex vivo and in vivo, which might represent a novel potential agent for the development of therapeutic drugs to treat ischemic diseases.