叶黄素对人RPE细胞氧化应激损伤的保护作用

Protective Effect of Lutein on Human RPE Cells from Oxidative Stress

目的:外源性给予过氧化氢(H_2O_2)诱导构建人视网膜色素上皮细胞(Retinal pigment epithelial,RPE)细胞氧化损伤模型,探究H_2O_2的最佳建模浓度,并探讨叶黄素对H_2O_2诱导人RPE细胞氧化损伤的保护作用。方法:本研究以人RPE细胞为实验对象。不同浓度H_2O_2(0、50、100、200、400、600μmol/L)处理RPE细胞1 h后,观察细胞形态的改变,并测定细胞生存率和细胞内ROS浓度进而确定H_2O_2的最佳建模浓度。不同剂量叶黄素(1、2.5、5、7.5、10μg/m L)预处理RPE细胞24h,随后给予100μmol/L H_2O_2作用1h,测定各组细胞生存率和细胞内活性氧(ROS)浓度,从而评价叶黄素对RPE细胞氧化损伤的作用。结果:H_2O_2作用后,随H_2O_2浓度的增加,RPE细胞生存率逐步下降;细胞内ROS浓度随H_2O_2的浓度增加而显著升高。与损伤对照组相比,各叶黄素处理组RPE细胞生存率显著升高,同时细胞内ROS浓度显著下降。结论:H_2O_2可导致RPE细胞出现氧化应激损伤,细胞ROS含量显著增加。叶黄素干预后可显著减缓H_2O_2诱导的氧化应激反应,提示其可通过提高RPE细胞的生存率、抑制细胞内ROS浓度,保护RPE细胞免受氧化损伤,从而对年龄相关性黄斑变性等眼部退行性疾病起到预防和减缓作用。

Objective To discuss the suitable concentration and explore the possible function of lutein against oxidative damage induced by H2O2 in cultured human RPE cells,we given Exogenous H2O2 to build the oxidative stress model. Method The object of experiment was human RPE cells. We using different concentrations of H2O2（ 0、50、100、200、400、600μmol/L） to deal with human RPE cells for 1 h. To ensure the suitable concentration of H2O2,we observed the changes of cell morphology by light microscopy and the density of intracellular ROS was also detected along with cell viability. Gradient concentrations of lutein solution（ 1、2. 5、5、7. 5、10μg/m L）were added into cultured human RPE cells to interfere with the oxidative stress for 24 h. After that the cellular viability and intracellular ROS were detected as well. Result After treating with H2O2 for 1 h,the cellular viability decreased along with the concentration of H2O2. On the contrary,the intracellular concentration of ROS and cell debris gradually increased. After pretreatment of lutein for 24 h,the cellular viability increased and meanwhile the intracellular concentration of ROS decreased compared with no-lutein control group. Conclusion The results suggested that exogenous H2O2 could induce RPE cells oxidative stress. The oxidative damage could be reduced after treating with lutein,showing that lutein has potential ability to prevent RPE cells from oxidative stress by increasing cellular viability and decreasing the ROS concentration. Thus,lutein played an important role in the age-related macular degeneration and other degenerative diseases.