姜黄素对人视网膜色素上皮细胞增殖和VEGF表达的影响

Effect of curcumin on proliferation and VEGF expression of human retinal pigment epithelium cells

目的探讨姜黄素对人视网膜色素上皮(hRPE)细胞增殖和血管内皮生长因子(VEGF)表达的影响。方法(1)采用不同浓度(5μg/mL、10μg/mL、20μg/mL、40μg/mL)的姜黄素干预hRPE细胞1 d、2 d、3 d、4 d、5 d、6 d,同时设置空白对照组(不给予姜黄素干预)。采用CCK-8法检测各时间点不同浓度姜黄素对hRPE细胞增殖的影响,采用流式细胞仪检测各时间点各组细胞周期时相分布情况。(2)将hRPE细胞分为姜黄素组和对照组,姜黄素组加入10μg/mL的姜黄素,对照组加入等体积培养液,采用实时荧光半定量PCR检测各组hRPE细胞VEGF mRNA的表达水平。结果(1)各浓度姜黄素对hRPE细胞均有抑制作用(均P<0.05),且随着药物浓度的增大、作用时间的延长,其抑制率呈升高趋势,整体呈现剂量和时间依赖性。(2)不同浓度姜黄素干预后,G_(0)/G_(1)期细胞比例均大于空白对照组,S期细胞比例和G_(2)/M期细胞比例均小于空白对照组(均P<0.05)。(3)姜黄素组hRPE细胞VEGF mRNA表达水平低于对照组(P<0.05)。结论姜黄素可以将hRPE细胞周期阻滞于S期从而抑制hRPE细胞的增殖,还可以降低hRPE细胞的VEGF表达水平。

Objective To explore the effect of curcumin on proliferation and vascular endothelial growth factor(VEGF)expression of human retinal pigment epithelium(hRPE)cells.Methods(1)hRPE cells were intervened by curcumin in different concentrations(5μg/mL,10μg/mL,20μg/mL,and 40μg/mL)for 1,2,3,4,5,and 6 days;simultaneously,blank control group was set(without intervention of curcumin).The effects of curcumin in different concentrations on hRPE cell proliferation at various time points were detected by employing the CCK-8 method.The cell cycle phase distribution of various groups at various time points was measured by using flow cytometry.(2)hRPE cells were assigned to curcumin group or control group,the curcumin group was added with 10μg/mL curcumin,and the control group was added with culture solution with equivalent volume.The VEGF mRNA expression of hRPE cells in various groups was detected by using real-time fluorescent semi-quantitative PCR.Results(1)There were inhibitory effects of curcumin in different concentrations on hRPE cells(all P<0.05),and with the increase of drug concentration and the extension of action time,the inhibitory rate gradually increased;overall,it is dose-and time-dependent.(2)After intervention with different concentrations of curcumin,the proportion of cells in G_(0)/G_(1) phase was larger than that in the blank control group,and the proportions of cells in S phase and G_(2)/M phase were smaller than those in the blank control group(all P<0.05).(3)The VEGF mRNA expression of hRPE cells in the curcumin group was lower than that in the control group(P<0.05).Conclusion Curcumin can inhibit the proliferation of hRPE cells by arresting hRPE cell cycle in S phase,and it can also decrease the expression of VEGF in hRPE cells.