紫外辐射诱导牡蛎细胞DNA损伤 单细胞凝胶电泳检测.docVIP

紫外辐射诱导牡蛎细胞DNA损伤 单细胞凝胶电泳检测 　　摘要: 以牡蛎为研究对象，建立了水产动物DNA损伤的单细胞凝胶电泳检测方法.分别以20W紫外灯距离30cm照射牡蛎血液细胞30、60和?120s，然后用单细胞凝胶电泳技术检测细胞DNA的损伤.结果表明：未照射的细胞未受损伤，在电场中核DNA几乎不泳动，染色后呈圆形的荧光团，无拖尾现象.紫外照射后细胞核DNA均不同程度受损，且拖尾率、尾长、尾矩、Olive尾矩等指标值均随着照射时间加长而增加，DNA损伤程度与紫外照射时间之间存在明显的时间效应关系.研究结果为我国海洋环境中辐射防护研究和辐射毒理学检测提供新的研究方法和思路. 　　关键词: 单细胞凝胶电泳;彗星实验;紫外辐射;DNA损伤;牡蛎 　　中图分类号:Q952.6 　　 文献标识码:A文章编号:1672-8513(2010)06-0400-04 　　 　　UV Radiation-Induced DNA Damage in Oyster Cells Measured by Single Cell Gel Electrophoresis 　　XIONG He?jian?1,MA Ying?2,WANG Lin?2,TANG Sen?ming?3 　　(1.College of Bioengineering, Jimei University，Xiamen 361021, China；2.College of Fisheries, Jimei University，Xiamen 361021, China；3.The Third Institute of Oceanography SOA， Xiamen 361021, China) 　　Abstract: In order to detect DNA damage induced by radiation in marine environments, the single cell gel electrophoresis (SCGE) technique for detecting DNA damage of aquatic animals was established. Oyster cells were exposed to a 200W ultraviolet (UV) lamp at a distance of 30cm for different times (0s, 30s, 60s and 120s), and the damage was measured by SCGE. The results showed that DNAs in the control group are free of damage, static in the electric field and appeared as a round fluorophore without any cometic tails. While those radiated DNAs are damaged to various degrees, and the four indices (Tail length, %DNA in tail, Tail moment, Olive tail moment and Tailing percent) used for assessing the damage increased with the radiation times. There existed a time?effect relationship between SCGE indices and UV radiation time. The results will provide new methods and clues in toxicology testing for radiation pollution in marine environments. 　　Key words: single cell gel electrophoresis; comet assay; UV radiation; DNA damage; oyster 　　 　　随着工农业的迅猛发展和人类活动的不断增加，环境污染日趋严重。环境污染对生物的影响已经成为人们??心的环境问题之一.研究发现，环境中某些物理（电离辐射等）和化学（重金属、农药、化学试剂等）因素会直接或间接作用于生物体DNA，干扰或破坏DNA的复制过程，引起DNA片断移位、易位和丢失等，进而引起动物的遗传损伤并影响其遗传行为［1］. 　　单细胞凝胶电泳（Single Cell Gel Eletrophoresis