不同方法检测微囊藻毒素-LR致DNA氧化损伤  

MCLR induce oxidative DNA damage as determined by different methods

作　　者：农清清[1] 蓝勇波 竹内亨[3] 张志勇[1] 何敏[1] 

机构地区：[1]广西医科大学公共卫生学院,广西南宁530021 [2]广西中医骨伤研究所,广西南宁530012 [3]日本鹿儿岛大学医齿学综合研究科环境医学研究室

出　　处：《中国热带医学》2008年第9期1505-1506,共2页China Tropical Medicine

基　　金：国家留学基金委资助项目(No20845014)

摘　　要：目的观察微囊藻毒素-LR(MCLR)对人肝癌细胞HepG2的DNA氧化损伤效应。方法应用高效液相色谱-电化学法(HPLC-ECD)和免疫酶染色法检测8-羟基脱氧鸟苷(8-OHdG)水平。结果用HPLC-ECD法分析,低、中剂量组8-OHdG水平显略有增高趋势,高剂量组(100μmol/L)8-OHdG水平显著高于空白对照组;用免疫酶染色法分析,各剂量组8-OHdG染色强度明显高于空白对照组,处理剂量与8-OHdG水平之间呈现剂量-反应关系。结论MCLR引起人肝癌细胞DNA氧化损伤,HPLC-ECD法与免疫酶染色法所测损伤的趋势一致。免疫酶染色法能够更灵敏地检测DNA氧化损伤,可应用于人类疾病研究与环境监测。Objective To study oxidative DNA damages in human hepatoma cell lines HepG2 treated with MCLR, as determined by different methods. Methods Oxidative DNA damages were determined by high - performance liquid chromatography with electrochemical detection （HPLC -ECD） and immunostaining of 8 - hydroxydeoxiguanosine （8 - OHdG）. Results MCLR at high （100umol/L） doses significantly increased 8 -OHdG levels, and the trend of increase in 8 -OHdG levels was detected in the low and medium doses by HPLC - ECD. The staining intensity of 8 - OHdG in cells treated with different doses of MCLR were stronger than that of untreated cells, and the dose - response relationship was founded by immunosraining of 8 - OHdG. Conclusion MCLR induced oxidative DNA damages in human hepatoma cell lines. The analysis resuits of the two methods are coincidence. Immunostaining of 8 - OHdG is reliable and sensitive method for detecting oxidative DNA damages and could be applied to the study of human diseases and environmental monitoring.

关 键 词：微囊藻毒素-LR DNA氧化损伤 8-羟基脱氧鸟苷 HPLC—ECD 免疫酶染色法 

分 类 号：R361[医药卫生—病理学] R994.6[医药卫生—基础医学]