Abstract
Purpose:
AMD is the most important cause of blindness in developed countries. The wet form of the disease is characterized by choroidal neovascularization, an abnormal growth of blood vessels from the choroid underneath the macula. The Resveratrol (3,5,4'-trihydroxystilbene) a naturally occurring phytoalexin, found in grapes and wine, has been reported to exert anti-oxidative and anti-inflammatory properties, processes known to be involved in the physiopathology of AMD. Here, we aimed to investigate the impact of Resveratrol treatment on processes associated with angiogenesis using the human umbilical vein endothelial cells (HUVEC).
Methods:
For all experiments HUVEC were treated 24h with Resveratrol. Optimal concentrations of Resveratrol were determined by apoptosis and cytotoxicity assays through measuring caspase-3 activity and LDH release respectively. Cell viability was assessed by MTT assay and hemocytometer-based trypan blue dye exclusion. A Propidium iodide and BrdU staining was used for cell cycle phase’s quantitation. Migration assay was performed on a confluent dish lesioned with a 200µl pipette tip.
Results:
Resveratrol induces in a dose dependent manner a reduction in cell numbers (% to Control with 15µM: 60,9±4.4% and 25µM: 48.8±4.4%) and BrdU incorporation (% to Control with 15µM: 75,8±8% and 25µM: 48.0±3.9%). Cell cycle analysis showed that this inhibition of proliferation rely on a reduced progression to the G2/M phase (Control: 6,9%, 25µM: 3,2%). Moreover, we showed that VEGF-induced cell migration was significantly inhibited by Resveratrol.
Conclusions:
This study shows inhibitory effects of Resveratrol on angiogenesis, using an in vitro model. It suggests that by its properties Resveratrol could be a good candidate for a clinical application in AMD. Other studies are still needed to confirm this hypothesis.