【摘要】 目的采用碱性单细胞凝胶电泳技术检测BDE-209染毒小鼠精子DNA的损伤,探讨BDE-209影响受精卵发育的可能机制。方法附睾尾取精法体外获得昆明小鼠精子,分别置于HTF培养基中。按照培养基中BDE-209终浓度的不同将实验分为5组:A组:0μg/ml;B组:2.5μg/ml;C组:5μg/ml;D组:10μg/ml;E组:20μg/ml。精子培养后,做单细胞凝胶电泳,再用彗星分析软件对细胞进行分析(SCGE),比较各组精子损伤的差异。结果在SCGE试验图片中,A组平均尾长为(1.15±1.27)μm。随着BDE-209浓度的增加,D组和E组,可见到部分细胞拖尾,平均尾长分别为(2.13±1.29)μm和(2.83.±2.46)μm,明显长于A组(P<0.01)。细胞尾部DNA的含量比也明显增加(P<0.01)。D组及E组的Olive尾矩、尾长/头长比A组长(P<0.01),C组的尾长/头长比A组大(P<0.05)。小鼠精子DNA损伤中各指标变化与BDE-209浓度的相关系数均大于0.9。结论外源性的BDE-209可造成小鼠精子DNA的损伤,导致小鼠精子DNA链的断裂。外源性BDE-209对小鼠精子DNA的损伤作用呈明显的剂量-效应关系。更多还原

【Abstract】 Objective To assess the DNA damage in mouse sperms induced by exogenous BDE-209 and explore the possible mechanism of BDE-209 in affecting normal zygote development.Methods Mouse sperms were harvested from the epididymal tail and suspended in HTF medium for a 90-min exposure to BDE-209 at varied concentrations of 0,2.5,5.0,10,and 20 μg/ml(groups A-E,respectively).After the exposure,the sperms were subjected to single-cell gel electrophoresis(SCGE) to assess the DNA damage.Results The tail length of the sperms averaged 1.15±1.27 μm in group A.Exposure to 10 and 20 μg/ml BDE-209 resulted in a significant lengthening of the sperm tails(2.13±1.29 μm and 2.83±2.46 μm,respectively,P<0.01) as well as increased DNA content in the tail of the cells(P<0.01).The Olive tail moment in group A was 0.270±0.322,and increased after BDE-209 exposure to 0.453±0.375 and 808±0.822 in groups D and E,respectively.The tail/head length ratio in groups C,D,and E(0.077±0.093,0.112±0.068,and 0.191±0.207) were significantly greater than that in group A(0.045±0.049).The DNA damage of the mouse sperms was directly correlated to the concentrations of BDE-209,with correlation coefficients all above 0.9.Conclusions Exogenous BDE-209 can cause mouse sperm DNA damage and lead to sperm DNA chain breakage,and this effect shows an obvious dose dependence.更多还原