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目的 研究60 Coγ射线照射人乳头状病毒永生化的人支气管上皮细胞 (BEP2D)产生的活性氧 (ROS)及其对DNA的氧化损伤 ,以及纳米硒的保护作用。方法 细胞上清中过氧化氢(H2 O2 )、超氧阴离子 (O 2 )以及羟自由基 (·OH)含量分别用辣根过氧化物酶介导的酚红氧化法、细胞色素C还原法和番红花红退色法进行测定 ;细胞内H2 O2 和O 2 分别用 2’ ,7’ 二氯荧光黄双乙酸盐 (DCFH DA)和氢化乙锭 (HE)标记 ,用流式细胞法测定荧光产物 2’ ,7’ 二氯荧光黄 (DCF)和溴乙锭 (EB)荧光强度 ;8 羟基脱氧鸟嘌呤 (OH8dG)含量用高压液相色谱结合电化学方法 (HPLC ECD)测定。结果 BEP2D细胞经不同剂量的60 Coγ射线照射后 ,细胞内及细胞上清中产生的ROS以及OH8dG含量均显著增加 ,并有较好的剂量效应关系。 1μmol/L的纳米硒能明显减少60 Coγ射线照射后BEP2D细胞产生的ROS及OH8dG水平。结论 60 Coγ射线照射能造成细胞的氧化损伤 ,纳米硒对60 Coγ射线照射所致细胞的氧化损伤有保护作用
Objective To investigate the effects of reactive oxygen species (ROS) produced by 60 Co γ -ray irradiated human immortalized human bronchial epithelial cells (BEP2D) on DNA oxidative damage and the protective effects of nano-selenium. Methods The contents of hydrogen peroxide (H2O2), superoxide anion (O2) and hydroxyl radical (· OH) in the supernatant of the cells were respectively measured by horseradish peroxidase-mediated phenol red oxidation and cytochrome C reduction And crocodile red fading method. The intracellular H2O2 and O2 were labeled with 2 ’, 7’ dichlorofluorescein diacetate (DCFH DA) and ethidium bromide (HE) respectively, and the fluorescence was measured by flow cytometry The fluorescence intensity of DCF and EB of the products 2 ’, 7’ was measured. The content of OH 8dG was determined by high pressure liquid chromatography coupled with electrochemical method (HPLC ECD). Results After irradiated with different doses of 60 Co γ-ray, BEP2D cells significantly increased the content of ROS and OH8dG in the cells and in the supernatant of cells, and had a good dose-response relationship. Nano-selenium at 1μmol / L could significantly reduce the levels of ROS and OH8dG produced by BEP2D cells after 60 Co γ -ray irradiation. Conclusion 60 Co γ-ray irradiation can cause oxidative damage of cells, and nano-selenium has a protective effect on the oxidative damage of cells induced by 60 Co γ -ray irradiation