洋葱总黄酮减轻过氧化氢诱导的视网膜色素上皮细胞氧化损伤

Total flavonoids of onion attenuate hydrogen peroxide-induced oxidative damage in retinal pigment epithelial cells

目的:探讨洋葱总黄酮(FO)对过氧化氢(H2O2)诱导的视网膜色素上皮细胞氧化损伤的影响。方法:将视网膜色素上皮ARPE-19细胞分成对照(control)组、H2O2组、低浓度FO(FO-L)+H2O2组、中浓度FO(FO-M)+H2O2组和高浓度FO(FO-H)+H2O2组。MTT法检测细胞活力,流式细胞术检测细胞凋亡。DCFH-DA荧光探针法检测细胞中活性氧簇(ROS)的水平,WST法检测超氧化物歧化酶(SOD)的活性,TBA法检测丙二醛(MDA)的含量,JC-1法检测细胞线粒体膜电位,Westernblot检测胞质中细胞色素C(Cyt-C)及细胞中cleaved caspase-3和cleaved caspase-9的蛋白水平。结果:H2O2降低ARPE-19细胞活力,升高细胞凋亡率和ROS水平,降低SOD活性,升高MDA含量,降低线粒体膜电位,升高胞质中Cyt C蛋白水平及细胞中cleaved caspase-3和cleaved caspase-9蛋白水平(P<0.05)。与H2O2组比较,FO-L+H2O2、FO-M+H2O2和FO-H+H2O2组的细胞活力逐渐升高,凋亡率逐渐降低,细胞中ROS水平逐渐降低,SOD活性逐渐升高,MDA含量逐渐降低,线粒体膜电位逐渐升高,胞质中Cyt C蛋白水平及细胞中cleaved caspase-3和cleaved caspase-9蛋白水平逐渐降低(P<0.05)。结论:洋葱总黄酮减轻H2O2诱导的视网膜色素上皮细胞氧化损伤。

AIM: To investigate the effects of total flavonoids of onion(FO) on hydrogen peroxide(H2O2)-induced oxidative damage in retinal pigment epithelial cells. METHODS: The retinal pigment epithelium ARPE-19 cells were divided into 5 groups: control group, H2O2 group(treated with H2O2), FO-L+H2O2 group(treated with H2O2 and low concentration of FO), FO-M+H2O2 group(treated with H2O2 and medium concentration of FO) and FO-H+H2O2 group(treated with H2O2 and high concentration of FO). The cell viability was measured by MTT assay. Apoptosis was analyzed by flow cytometry. DCFH-DA staining was used to detect reactive oxygen species(ROS) level in the cells. WST assay was used to detect superoxide dismutase(SOD) activity. The content of malonaldehyde(MDA) was measured by TBA method. Mitochondrial membrane potential was analyzed by JC-1 staining. The protein levels of cytochrome C(Cyt C) in the cytoplasm, and cleaved caspase-3 and cleaved caspase-9 in the cells were determined by Western blot. RESULTS: Treatment with H2O2 decreased ARPE-19 cell viability, increased the apoptotic rate and the level of ROS in the cells, decreased SOD activity, increased the content of MDA, decreased mitochondrial membrane potential, and increased the protein levels of Cyt C in the cytoplasm and cleaved caspase-3 and cleaved caspase-9 in the cells(P<0.05). Compared with H2O2 group, the cell viability in FO-L+H2O2 group, FO-M+H2O2 group and FO-H+H2O2 group was increased, the apoptotic rates were decreased, the levels of ROS were decreased, SOD activity was increased, the content of MDA was decreased, mitochondrial membrane potential was increased, the protein level of Cyt C was decreased in the cytoplasm, and the protein levels of cleaved caspase-3 and cleaved caspase-9 protein in the cells were decreased gradually(P<0.05). CONCLUSION: Total flavonoids of onion reduce H2O2-induced oxidative damage in retinal pigment epithelial cells.