摘要
目的:探究虾青素对蓝光发光二极管(light-emitting diodes,LED)诱导ARPE-19细胞氧化应激损伤的保护作用及其机制。方法:采用不同浓度的虾青素预处理细胞1 h后,蓝光LED下光氧化损伤诱导24 h。噻唑蓝法测定细胞活力,乳酸脱氢酶检测法分析细胞毒性,2’,7’-二氯二氢荧光素二乙酸酯荧光探针法测定细胞内活性氧(reactive oxygen species,ROS)的水平,JC-1荧光探针法测定细胞内线粒体膜电位的变化,利用酶学相关试剂盒测定细胞内源性抗氧化酶活力,实时荧光定量聚合酶链式反应法测定细胞内Ⅱ相解毒基因的转录表达水平,Western blot法测定细胞内核因子E2相关因子2(nuclear factor erythroid-2-related factor 2,Nrf2)的核蛋白表达水平。结果:虾青素(5、10μmol/L和20μmol/L)预处理以浓度依赖的方式抑制蓝光LED诱导的ARPE-19细胞活力降低,缓解细胞毒性;虾青素预处理能够减少细胞内ROS的产生,稳定线粒体膜电位,提高内源性抗氧化酶活力。此外,虾青素诱导了Nrf2的核转位,增加了抗氧化酶和Ⅱ相解毒基因的表达,从而保护ARPE-19细胞免受蓝光LED诱导的氧化应激损伤。结论:虾青素通过诱导Nrf2的核转位,促进抗氧化酶和Ⅱ相解毒基因的表达水平升高从而保护ARPE-19细胞免受蓝光LED诱导的氧化应激损伤。
Objective:To investigate the protective effect of astaxanthin on oxidative stress injury induced by blue lightemitting diode(LED)in ARPE-19 cells and its underlying mechanism.Methods:The cells were pretreated with different concentrations of astaxanthin for 1 h followed by exposure to blue light LED for 24 h to induce photooxidative damage.The cell viability was determined by the MTT method,the cytotoxicity was determined by the lactate dehydrogenase(LDH)assay,the level of intracellular reactive oxygen species(ROS)was measured using 6-carboxy-2’,7’-dichorodihydrofluorescein diacetate(DCFH-DA)as a fluorescence probe,the change of intracellular mitochondrial membrane potential was measured using the fluorescence probe JC-1,and the activity of endogenous antioxidant enzymes was determined by commercial kits.The transcriptional expression level of phaseⅡdetoxification genes was measured by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR),and the nuclear protein expression level of nuclear factor erythroid-related factor 2(Nrf2)was measured by Western blot.Results:Astaxanthin(at 5,10 and 20μmol/L)pretreatment inhibited the decrease in the cell viability induced by blue light LED in a concentration-dependent manner,alleviated the cytotoxicity,reduced the production of intracellular ROS,stabilized the mitochondrial membrane potential and increased the activity of endogenous antioxidant enzymes.In addition,astaxanthin induced nuclear translocation of Nrf2 and increased the expression of antioxidant enzymes and phaseⅡdetoxification genes,thus protecting ARPE-19 cells from oxidative stress induced by blue light LED.Conclusion:Astaxanthin protects ARPE-19 cells from oxidative stress induced by blue light LED by inducing nuclear translocation of Nrf2 and promoting the expression of antioxidant enzymes and phaseⅡdetoxification genes.
作者
刘莹
马宁
刘垚杰
郭雅图
李赫宇
单艳琴
陈云霞
王浩
LIU Ying;MA Ning;LIU Yaojie;GUO Yatu;LI Heyu;SHAN Yanqin;CHEN Yunxia;WANG Hao(College of Food Science and Engineering,Tianjin University of Science and Technology,Tianjin 300457,China;Graduate School of Chengde Medical University,Chengde 067000,China;Cangzhou People’s Hospital,Cangzhou 061000,China;Tianjin Eye Hospital,Tianjin 300020,China;Tianjin Ubasio Biotechnology Group Co.,Ltd.,Tianjin 300457,China;Jiangsu Natural Food Co.,Ltd.,Taizhou 225700,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2021年第21期128-136,共9页
Food Science
基金
天津市科技计划项目(17KPHDSF00120)
河北省研究生创新资助项目(CXZZSS2019138)。
