三种抗氧化剂对氯化镉所致河蟹精子DNA损伤的保护作用  

Protective effects of 3 kinds of antioxidants on CdCl_2-induced Eriocheir sinensis sperm DNA damage

作　　者：范丽君[1] 曲迪[1] 贾林芝[1] 李艳东[1] 赵云龙[1] 王群[1] 

机构地区：[1]华东师范大学生命科学学院,上海200062

出　　处：《水产学报》2007年第5期561-567,共7页Journal of Fisheries of China

基　　金：国家自然科学基金项目(30300265)

摘　　要：采用单细胞凝胶电泳技术与CASP软件检测和分析了3种抗氧化剂(VC,VE和Zn2SO4)对氯化镉所致河蟹离体精子DNA损伤保护作用的量效关系。以5mmol.L-1氯化镉处理河蟹精子DNA,表征DNA损伤程度的4个参数(L tail,TailDNA,TM,OTM)值与空白组相比显著增加,表明该氯化镉浓度下已对河蟹精子的DNA造成严重损伤。在此基础上,将河蟹精子加入到分别含有不同浓度抗氧化剂的氯化镉(5mmol.L-1)溶液中,研究不同浓度抗氧化剂对精子DNA的保护作用。其中VC的浓度分别为:400μmol.L-1,1mmol.L-1,3mmol.L-1,5mmol.L-1;VE的浓度分别为:50μmol.L-1,100μmol.L-1,200μmol.L-1,400μmol.L-1;Zn2+的浓度分别为:400μg.mL-1,1mg.mL-1,2mg.mL-1,4mg.mL-1。结果发现,上述3种抗氧化保护剂均对精子DNA具有一定的保护效果,其中VE和Zn2+添加组均随其浓度的增加,保护作用不断增强,当VE达到400μmol.L-1,Zn2+达到4mg.mL-1时,均表现出很好的保护效果;然而,VC在低浓度时保护作用随其浓度增加而加强,至3mmol.L-1时细胞拖尾率最低,即保护效果最佳,但在5mmol.L-1时细胞拖尾率又明显增加,即精子DNA又受到严重损伤,表明VC对精子DNA的保护作用具有一定的浓度范围。此外,与VC和Zn2+的保护作用相比,VE在400μmol.L-1浓度时表现出更好的保护效果。To investigate the protective role of three antioxidants-vitamin C, vitamin E and Zn （Zn2SO4 · 7H2O） in CdCl2-induced Eriocheir sinensis sperm DNA damage, single cell gel electrophoresis was used to detect the relationship between the contents of antioxidants and DNA damage. Four parameters, L tail （ Tail length in arbitrary units）, Tail DNA （relative % of DNA in the comet tail）, TM （tail moment means integrated value of DNA density multiplied by the migration distance, L tail × Tail DNA） and OTM （ Olive tail moment means the product of the distance between the center of gravity of the head and the center of gravity of the tail and percent tail DNA）, were used to evaluate the DNA damage of the Eriocheir sinensis sperms which were treated by 5 mmol · L^-1 CdCl2. The result showed that the four parameters all increased remarkably and the sperm DNA was seriously damaged by CdCl2. After that the protective effects of 3 kinds of antioxidants on CdCl2-induced DNA damage were investigated. The contents of Vc were 400 μmol · L^-1,1 mmol · L^-1, 3 mmol · L^-1 and 5 mmol · L^-1 ;that of VE were 50 μmol · L^-1,100 μmol · L-1,200 μmol · L^-1 and 400 μmol · L^-1 ; and that of Zn were 400 μg · mL^-1,1 mg · mL^-1,2 mg · mL^-1 and 4 mg · mL^-1, respectively. The result showed that all these three antioxidants can protect Eriocheir sinensis sperm DNA from oxidizing. The bigger the content of VE and Zn, the stronger the protective effect was. When the content of Vc was 3 mmol · L^-1, the protective effect was the best and the ratio of cell drag was comparatively low. But when the content of that was 5 mmol · L^-1 , the ratio of cell drag increased and the DNA damage became more serious. So it was concluded that the content of Vc which could protect the sperm DNA from damaging was limited. Whereas the protective effect of VE and Zn had no limit of the content. Furthermore, by contrast to Vc and Zn, VE appeared better protective effects with concentration of 400 μmol · L^-1. Vitamin E and

关 键 词：维生素C 维生素E 硫酸锌 氯化镉 彗星实验 DNA损伤 单细胞凝胶电泳技术 

分 类 号：S917[农业科学—水产科学]