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Dariusz Rokicki, Robert Zdanowski, Slawomir Lewicki, Monika Lesniak, Milena Suska, Ewa Skopinska-Rozewska, Piotr Skopinski; In vitro effects of Resveratrol and Resvega to Endothelial Murine Cells Culture . Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4298.
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© ARVO (1962-2015); The Authors (2016-present)
Pathological angiogenesis plays an essential role in the pathomechanism of numerous disorders, including e.g.exudative age-related macular degeneration (AMD) or proliferative diabetic retinopathy. Since the crucial role is played by the angiogenesis in these conditions, there has been increased interest in evaluating potential anti-angiogenic agents in their management. The aim of our study was to assess in vitro the influence of resveratrol and resveratrol based formula (Resvega) on proliferation, migration and invasion of immortalized murine endothelial cell line from peripheral lymph node HEC clone a10.
Proliferation of endothelial cells was estimated using MTT, and neutral red assay (NR). MTT assay evaluates viability based on the conversion of MTT into formazan crystals by living cells, which determines mitochondrial activity, whereas NR uptake is directly proportional to the number of viable cells actively accumulating dye neutral red in lysosomes.<br /> In addition, cell migration and cell invasion assays were examined using BD FluoroBlok™ fluorescence blocking membranes in 24-well plates. Resveratrol (1, 5, 10, 50 or 100 µM, Sigma Aldrich) or Resvega (1, 5, 10, 50 or 100 µM, Laboratoires Thea) were added to the lower chamber. As a control was used culture medium without resveratrol or Resvega, and as a negative control - FBS-free culture medium. Statistical analysis of the results was performed with two-way (proliferation) and one-way (migration and invasiveness) ANOVA with Tukey’s Multiple Comparison Test (GraphPadPrism).
Resveratrol and Resvega were shown to inhibit the proliferation of the endothelial cells in MTT (concentration-dependently) and NR (at highest concentrations only) assays, and both agents at the concentration of 50 μM significantly inhibited migration of endothelial cells. A concentration-dependent decrease in invasion potential of endothelial cells incubated with resveratrol or Resvega 10 μM and 50 μM was detected.
These promising in vitro results might encourage investigators to testify efficacy and safety of resveratrol based products as e.g. Resvega, more extensively in the clinical practice.
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