丁苯酞通过Nrf2/ARE信号通路保护高糖诱导的视网膜色素上皮细胞免受氧化损伤
Butylphthalide Protects High Glucose-Induced Retinal Pigment Epithelium Cells from Oxidative Damage through the Nrf2/ARE Signaling Pathway

目的：研究丁苯酞对高糖诱导的视网膜色素上皮(RPE)细胞氧化应激损伤的保护作用，并探究其具体作用机制，为糖尿病视网膜病变的治疗提供新的思路。方法：应用CCK-8实验筛选丁苯酞安全浓度，将细胞分为以下3组：正常组(低糖DMEM，糖浓度5.5 mM)，高糖组(高糖DMEM，糖浓度25 mM)，丁苯酞组(丁苯酞 + 高糖DMEM)，并检测细胞活力。采用细胞划痕实验检测细胞迁移能力，活性氧试剂盒测定细胞内活性氧(ROS)表达量，Western Blot检测Nrf2/ARE信号通路相关蛋白表达。结果：丁苯酞浓度 ≤ 100 μM时，对细胞存活率无明显影响(P > 0.05)，后续丁苯酞实验浓度为100 μM。高糖组细胞存活率明显低于正常组(P < 0.001)，丁苯酞组细胞存活率明显高于高糖组(P < 0.001)，但较正常组降低(P < 0.001)。高糖组细胞迁移率明显高于正常组(P < 0.001)，丁苯酞组较高糖组细胞迁移率明显降低(P < 0.001)，但高于正常组(P < 0.001)。高糖组细胞内ROS水平较正常组明显升高(P < 0.001)，丁苯酞组较高糖组ROS水平降低(P < 0.001)，但较正常组ROS水平升高(P < 0.001)。高糖组细胞内Nrf2、HO-1、NQO1蛋白表达均较正常组升高(P均 < 0.001)，丁苯酞处理后Nrf2、HO-1、NQO1蛋白均进一步显著升高(P < 0.001, P < 0.001, P < 0.01)。结论：丁苯酞对高糖诱导的RPE细胞具有保护作用，其机制可能与激活Nrf2/ARE通路、抑制氧化应激密切相关。
Purpose: The aim of this study was to assess the protective effect of butylphthalide on oxidative stress damage of retinal pigment epithelium (RPE) cells induced by high glucose, and explore its specific mechanism, providing a new idea for the treatment of diabetic retinopathy. Methods: The safe concentration of butylphthalide was screened by Cell Counting Kit-8 (CCK-8) assay, and the cells were divided into three groups. The groups included a normal group, in which cells were cultured in low glucose DMEM medium with a concentration of 5.5 mM; a high glucose group, in which cells were cultured in high glucose DMEM medium with a concentration of 25 mM; a butylphthalide group, in which cells were cultured in butylphthalide and high glucose DMEM medium. Cell viability was also measured. The cell migration ability was detected by cell scratching assay. The expression of intracellular reactive oxygen species (ROS) was determined by reactive oxygen species kit. The expression of Nrf2/ARE signaling pathway related proteins was detected by Western Blot. Results: When the concentration of butylphthalide was less than 100 μM, the cell survival rate was not sig-nificantly affected (P > 0.05), and the concentration of butylphthalide was 100 μM in our study. The cell survival rate in the high glucose group was significantly lower than the normal group (P < 0.001). The cell survival rate in the butylphthalide group was significantly higher than the high glucose group (P < 0.001), but was lower than the normal group (P < 0.001). Cell mobility in the high glucose group was significantly higher than the normal group (P < 0.001). Cell mobility in the butylphthalide group was significantly lower than the high glucose group (P < 0.001), but higher than the normal group (P < 0.001). The intracellular ROS level in the high