Effects Of Resveratrol On The Proliferation Of Human Retinal Pigment Epithelial Cells In Vitro

Objective To investigate the effects of resveratrol(Res) on the proliferation of ARPE-19 cells and to explore the possible mechanisms preliminary.Methods The ARPE-19 cells were recovered and cultured. The biological characteristics was observed under inverted microscope. After ARPE-19 cells were treated with Res at concentrations of 0, 50, 100, 150, 200, 300 μmol/L respectively for 24 h, 48 h and 72 h, the effects of Res on the proliferation of the cells were tested by cell count kit-8(CCK-8) assay. The ARPE-19 cells were treated with Res at concentrations of 0, 100, 150, 200 μmol/L for 48 h. The effects of Res on the cell cycle and apoptosis were detected by flow cytometry with PI staining and Annexin V-FITC/PI, respectively. The m RNA expression levels of proliferation cell nuclear antigen(PCNA), P21 and P27 were determined by quantitative reverse transcription PCR(q RT-PCR). The protein expression of PCNA, P21 and P27 were detected by immunofluorescent assay and western blot.Results Most ARPE-19 cells started to grow adhering to the wall within 24 h after subculturing. The cells shaped as round at first, and then spread gradually and became polygons. A large amount of pigment granules were found around the transparent cell nucleus. The results of CCK-8 assay showed that Res inhibited the proliferation of ARPE-19 cells in a time-and dose-dependent manner. The treatment with Res for 48 h resulted in an arrest of cell cycle at S phase without increasing cell apoptosis. Res decreased the m RNA and protein expression of PCNA, and increased the m RNA and protein expression of P21 and P27 with the concentration gradually increasing.Conclusion Res inhibits the proliferation of ARPE-19 cells and induces the cell cycle arrest at S phase. The mechanism may be related to up-regulation of P21 and P27, and down-regulation of PCNA.