铜暴露下赤子爱胜蚓(Eisenia foetida)活体基因的损伤研究Detection of DNA Damage in Earthworm(Eisenia foetida)in Vivo Exposure to Copper Ion
卜元卿;骆永明;滕应;李振高;
摘要(Abstract)：
通过碱性单细胞凝胶电泳法研究了Cu2+暴露剂量对赤子爱胜蚓(Eiseniafoetida)活体基因损伤的动态影响.结果显示:不添加Cu2+的对照组和添加Cu2+的处理组蚯蚓体腔细胞尾部DNA含量和尾长均呈非正态分布(p<0.05);在暴露72h时,125mg·L-1Cu2+处理组尾部DNA含量值最大,为41.44%,100mg·L-1Cu2+处理组尾长值最大,为33.79μm;随着Cu2+暴露剂量的增加,尾部DNA含量和尾长损伤频率增加;对照组和处理组的尾部DNA含量和尾长之间均存在显著性差异(p<0.05),Spearman非参数相关分析表明,尾部DNA百分含量和尾长之间呈显著相关(p<0.01,n=21),Cu2+暴露浓度与尾部DNA百分含量、尾长具有良好的剂量-构效关系(p<0.01).在125mg·L-1Cu2+浓度下暴露72h时蚯蚓的基因损伤程度达到最大,损伤程度为3级.可见,蚯蚓DNA生物标志物是重金属污染基因毒理诊断的重要指标,碱性SCGE试验是检测Cu2+暴露对赤子爱胜蚓活体基因损伤的有效手段.
关键词(KeyWords)： 碱性单细胞凝胶电泳;Cu2+;蚯蚓;基因损伤;生物标志物
基金项目(Foundation): 国家自然科学基金重点项目(No.40432005);; 国家重点基础研究发展规划项目(No.2002CB410810/9)
作者(Author): 卜元卿;骆永明;滕应;李振高;
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DOI:
参考文献(References)：
- Biri A,Civelek E,Karahalil B,Sardas S.2002.Assessment ofDNA damage in women using assessment of DNA damage[J].Mutation Research,521(1-2):113-119
- Bengtsson G,Ek H,Rundgren S.1992.Evolutionary response ofearthworms to long term metal exposure[J].Oikos,63(2):289-297
- Cerda H,Delincée H,Haine H,Rupp H.1997.The DNA‘comet assay’as a rapid screening technique to control irradiatedfood[J].Mutation Research,375(2):167-181
- Chen Y,Wang Z J.2005.Detection of genotoxicity of soilpollution to earthworm in vivo exposure by comet assay[J].Acta Pedologica Sinica,42(4):577-583(in Chinese)
- Collins A R.2004.The comet assay for DNA damage andrepair:Principles,applications,and limitations[J].Mol.Biotechnol,26(3):249-261
- Conder J M,Seals L D,Lanno R P.2002.Method fordetermining toxicologically relevant cadmium residues in theearthworm Eisenia fetida[J].Chemosphere,49(1):1-7
- Escher B I,Hermens J L M.2002.Modes of action inecotoxicology:Their role in body burdens,species sensitivity,QSARs,and mixture effects[J].Environ Sci Technol,36(20):4201-4217
- Friis K,Damgaard C,Holmstrup M.2004.Sublethal soil copperconcentrations increase mortality in the earthworm Aporrectodeacaliginosa during drought[J].Ecotoxicology and EnvironmentalSafety,57(1):65-73
- Hankard P K,Svendsen C,Wright J,Wienberg C,Fishwick SK,Spurgeon D J,Weeks J M.2004.Biological assessment ofcontaminated land using earthworm biomarkers in support ofchemical analysis[J].Science of the Total Environment,330(1-3):9-20
- Hayashi M,Miyane K,Hirooka T,Endoh D,Higuchi H,Nagahata H,Nakayama K,Kon Y,Okui T.2004.Inhibitoryeffects of trientine,a copper-chelating agent,on induction ofDNA strand breaks in hepatic cells of Long-Evans Cinnamon rats[J].Biochimica et Biophysica Acta(BBA)-General Subjects,1674(3):312-318
- Lee R F,Steinert S.2003.Use of the single cell gelelectrophoresis/comet assay for detecting DNA damage in aquatic(marine and freshwater)animals[J].Mutation Research,544(1):43-64
- Lukkari T,Taavitsainen M,Soimasuo M,Oikari A,Haimi J.2004.Biomarker responses of the earthworm Aporrectodeatuberculata to copper and zinc exposure:Differences betweenpopulations with and without earlier metal exposure[J].Environmental Pollution,129(3):377-386
- M!ller P.2005.Assessment of reference values for DNA damage detected by the comet assay in human blood cell DNA[J].Mutation Research,612(2):84-104
- Ma W.2005.Critical body residues(CBRs)for ecotoxicologicalsoil quality assessment:Copper in earthworms[J].Soil Biologyand Biochemistry,37(3):561-568
- Moretti M,Marcarelli M,Villarini M,Fatigoni C,Scassellati-Sforzolini G,Pasquini R.2002.In vitro testing for genotoxicity ofthe herbicide terbutryn:Cytogenetic and primary DNA damage[J].Toxicol In Vitro,16(1):81-88
- Naga Raju G J,Sarita P,Ravi Kumar M,Ramana Murty G AV,Seetharami Reddy B,Lakshminarayana S,Vijayan V,RamaLakshmi P V B,Gavarasana S,Bhuloka Reddy S.2006.Traceelemental correlation study in malignant and normal breast tissueby PIXE technique[J].Nuclear Instruments and Methods inPhysics Research B,247(2):361-367
- OECD.1984.Guideline for testing chemicals,No.207.Earthworm,acute toxicity tests[S].Paris:Organisation forEconomic Cooperation and Development
- "stling O,Johanson K J.1987.Bleomycin,in contrast to gammairradiation,induces extreme variation of DNA strand breakagefrom cell to cell[J].International Journal of Radiation Biology,52(5):683-691
- Reinecke S A,Reinecke A J.2004.The comet assay asbiomarker of heavy metal genotoxicity in earthworms[J].Archives of Environment Contamination and Toxicology,46(2):208-215
- Singh N P,McCoy M T,Tice R R,Schneider E L.1988.Asimple technique for quantitation of low levels of DNA damage inindividual cells[J].Experimental Cell Research,175(1):184-191
- Srinivasan A,Lehmler H J,Roberson L W,Ludewig G.2001.Production of DNA strand breaks in vitro and reactive oxygenspecies in vitro and in HL-60cells by PCB metabolites[J].Toxicological Sciences,60(1):92-102
- Van Gestel C A M,Van Brummelen T C.1996.Incorporation ofthe biomarker concept in ecotoxicology calls for a redefinition ofterms[J].Ecotoxicology,5(4):217-225
- 陈颖,王子健.2005.用彗星试验检测土壤污染对蚯蚓活体基因损伤[J].土壤学报,42(4):577-583◆