摘要
目的探讨番茄红素对体外培养的人视网膜色素上皮细胞(retinal pigment epithelial cells,RPE)氧化损伤的保护作用及可能机制。方法 RPE细胞传代培养,过氧化氢(H_2O_2)处理制备氧化损伤模型。细胞随机分为阴性对照组(以正常培养液培养),氧化损伤组(100μmol·L^(-1)的H_2O_2作用12h),番茄红素低剂量组(1×10-6mol·L^(-1)番茄红素孵育24 h后,加入H_2O_2作用12 h)、番茄红素中剂量组((1×10-5mol·L^(-1)番茄红素孵育24 h后,加入H_2O_2作用12 h)和番茄红素高剂量组((1×10-4mol·L^(-1)番茄红素孵育24 h后,加入H_2O_2作用12 h),MTT比色法检测细胞存活率,流式细胞仪检测细胞活性氧(ROS)水平,DCFH-DA荧光探针检测胞内ROS变化,取培养上清液测定超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH)活性及丙二醛(MDA)的含量。结果番茄红素明显抑制了H_2O_2诱导的细胞活性的下降,减少了H_2O_2所致RPE细胞内ROS的生成,引起RPE细胞内SOD、GSH-Px水平升高及MDA水平下降。结论番茄红素通过提高细胞内SOD、GSH-Px含量,降低细胞内MDA含量发挥其较强的抗氧化作用。
Objective To discuss the protective effects of lycopene in vitro cultured human retinal pigment epithelial cells（ RPE） induced by oxidative damage and its possible mechanisms.Methods RPE cells were passaged and subcultured,treated with hydrogen peroxide（ H_2O_2）for preparation of oxidative damage model. Cells were randomly divided into negative control group（ normal cultured medium）,oxidative injury group（ 100 μmol·L^（-1）H_2O_2 treated for 12h）,low-dose lycopene group（ 1 × 10^-6mol · L^（-1）lycopene incubated for 24 h,then H_2O_2 treated for 12 h）,middle-dose lycopene group（ 1 × 10^-5mol·L^（-1）lycopene incubated for 24 h,then H_2O_2 treated for 12 h） and high-dose lycopene group（ 1 × 10^-4mol·L^（-1）lycopene incubated for 24 h,then H_2O_2 treated for 12 h）. Cell viability was detected by MTT assay,intracellular reactive oxygen species（ ROS） level was determined by flow cytometry,changes in intracellular ROS,superoxide dismutase（ SOD）,glutathione peroxidase（ GSH） activity were detected by DCFH-DA fluorescent probe and malondialdehyde（ MDA） content were measured in culture supernatant. Results Lycopene inhibited H_2O_2-induced cell activity decrease,reduced H_2O_2-induced ROS generation within the RPE cells,increased SOD,GSH-Px level and decreased MDA level. Conclusion Lycopene play its strong antioxidant effect by increasing the intracellular SOD,GSH-Px content and decreasing MDA content.
出处
《哈尔滨医科大学学报》
CAS
2016年第3期201-204,共4页
Journal of Harbin Medical University
基金
黑龙江省博士后资助经费(LBH-Z14161)
关键词
番茄红素
RPE细胞
氧化损伤
活性氧
年龄相关性黄斑变性
lycopene
retinal pigment epithelial cells
oxidative stress
reactive oxygen species
age-related macular degeneration
