白藜芦醇对视网膜色素上皮细胞增殖的影响

摘要

AIM:To investigate the effects of resveratrol ( Res) on the proliferation of ARPE-19 cells and to ex-plore the possible mechanisms.METHODS:After ARPE-19 cells were treated with Res at concentrations of 0, 50, 100, 150, 200 and 300 μmol/L for 24 h, 48 h and 72 h, the effects of Res on the proliferation of the cells were tested by CCK-8 assay.The ARPE-19 cells were treated with Res at concentrations of 0, 100, 150 and 200 μmol/L for 48 h.The effects of Res on the cell cycle and apoptosis were detected by flow cytometry with Annexin V-FITC/PI staining.The protein expression of proliferating cell nuclear antigen (PCNA) was detected by immunofluorescent assay.The mRNA expression of PCNA, P21 and P27 was determined by real-time PCR.RESULTS:The results of CCK-8 assay showed that Res inhibited the prolifera-tion 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 inhibited the protein expression of PCNA in ARPE-19 cells.The re-sults of real-time PCR showed that Res increased the mRNA expression of P21 and P27, and decreased the mRNA expres-sion of PCNA.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.%目的：探讨白藜芦醇（ resveratrol，Res）对视网膜色素上皮细胞增殖的影响，并初步探讨其作用机制。方法：用0、50、100、150、200和300μmol/L Res作用于ARPE-19细胞24、48和72 h后，CCK-8法检测Res对ARPE-19细胞增殖的影响，0、100、150和200μmol/L Res作用ARPE-19细胞48 h，PI单染流式细胞术检测细胞周期，Annexin V-FITC/PI双染流式细胞术检测细胞凋亡，免疫荧光化学法检测PCNA蛋白表达，实时荧光定量PCR （real-time PCR）法检测PCNA、P21和P27的mRNA表达水平。结果： CCK-8结果显示，Res抑制ARPE-19细胞增殖，呈时间和剂量依赖性；流式细胞术显示，Res使S期细胞百分比显著上升且各组凋亡率没有区别；免疫荧光显示， Res抑制PCNA蛋白表达，荧光减弱；real-time PCR显示， Res浓度依赖性地抑制PCNA的mRNA表达，而诱导表达P21和P27的mRNA。结论：Res能抑制ARPE-19细胞的增殖，使细胞阻滞在S期，其机制可能与白藜芦醇诱导P21与P27的mRNA表达、抑制PCNA的mRNA表达有关。