^(60)Coγ射线对BEP2D细胞氧化损伤的研究  

Oxidative damage of BEP2D cells irradiated ^(60)Coγ rays

作　　者：朱茂祥[1] 杨陟华[1] 龚诒芬[1] 徐勇 

机构地区：[1]北京放射医学研究所,100850 [2]美国印第安那大学医学院药理毒理研究室

出　　处：《中华放射医学与防护杂志》2000年第4期248-251,共4页Chinese Journal of Radiological Medicine and Protection

摘　　要：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60Objective\ To study reactive oxygen species (ROS) productions and oxidative DNA damage in HPV\|16 immortalized human bronchial epithelial cells(BEP2D) irradiated with 60 Co γ\|rays. Methods\ The measurement of extracellular superoxide anions (O  2) was based on the reduction of ferricytochrome C as assayed by the increase in its absorbance at 550 nm.Quantitation of extracellular hydrogen peroxides (H 2O 2) was based on the horseradish peroxidase\|dependent oxidation of phenol red which is assayed by its increased absorbance at 620 nm.The determination of extracellular hydroxyl radicals (·OH) was based on decoloration of safranine T as assayed by the decrease in its absorbance at 520 nm.Ethidium bromide and 2',7'\|dichlorofluorescein,fluorescent products of the membrane\|permeable dyes\|hydroethine and 2',7'\|dichloroflurescin diacetate,were used to monitor the intracellular production of H 2O 2 and O  2 respectively by flow cytometry.8\|hydroxydeoxygunosine (OH8dG) was examined with HPLC\|ECD from extracted DNA. Results\ The ROS production including H 2O 2,O  2 and ·OH,and DNA adduct OH8dG in 60 Co γ\|rays\|irradiated BEP2D cells increased remarkably,and these increments could be inhibited effectively by 1 μmol/L of selenium. Conclusion\ 60 Co γ\|irradiation causes oxidative damage to BEP2D cells,which can be protected by selenium.\;

关 键 词：钴60γ射线 BEP2D细胞 氧化损伤 研究 肿瘤 

分 类 号：R14[医药卫生—公共卫生与预防医学]