Bian Q, Xu L C, Wang S L, Xia Y K, Tan L F, Chen J F, Song L, Chang H C, Wang X R
Institute of Toxicology, Nanjing Medical University, Nanjing 210029, China.
Occup Environ Med. 2004 Dec;61(12):999-1005. doi: 10.1136/oem.2004.014597.
To determine sperm nuclear DNA integrity and to investigate the relation between fenvalerate (FE) exposure and spermatozoa DNA damage.
Sperm DNA fragmentation was detected by a modified alkaline single cell gel electrophoresis (Comet) assay and a terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) assay. The olive tail moment (OTM) and percentage tail DNA were measured by the Comet assay, and cell positive percentage was measured by the TUNEL assay for DNA damage evaluation.
The DNA integrity of spermatozoa of external and internal control groups were both significantly greater than that of the FE exposed group. The median value of tail DNA percentage in the exposure group was 11.30, which was significantly higher than 5.60 in the internal control group and 5.10 in the external control group. The median value of OTM was 3.80 in the exposure group, significantly higher than 1.50 in the internal control group and 2.00 in the external control group. Mean cell positive was 31.2% in the exposure group, significantly higher than 17.4% in the internal control and 19.6% in the external control groups. Cell positive (%) was significantly correlated with tail DNA percentage and with OTM of whole subjects (n = 63).
Results showed that occupational FE exposure is associated with an increase in sperm DNA damage. A combination of the Comet and TUNEL assays would offer more comprehensive information for a better understanding of sperm DNA damage, and the biological significance of sperm DNA damage in sperm function and male infertility.
确定精子核DNA完整性,并研究氰戊菊酯(FE)暴露与精子DNA损伤之间的关系。
采用改良的碱性单细胞凝胶电泳(彗星试验)和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)试验检测精子DNA片段化。通过彗星试验测量橄榄尾矩(OTM)和尾DNA百分比,通过TUNEL试验测量细胞阳性百分比以评估DNA损伤。
外部和内部对照组精子的DNA完整性均显著高于FE暴露组。暴露组尾DNA百分比的中位数为11.30,显著高于内部对照组的5.60和外部对照组的5.10。暴露组OTM的中位数为3.80,显著高于内部对照组的1.50和外部对照组的2.00。暴露组平均细胞阳性率为31.2%,显著高于内部对照组的17.4%和外部对照组的19.6%。在所有受试者(n = 63)中,细胞阳性率(%)与尾DNA百分比和OTM显著相关。
结果表明职业性FE暴露与精子DNA损伤增加有关。彗星试验和TUNEL试验相结合可为更好地理解精子DNA损伤以及精子DNA损伤在精子功能和男性不育中的生物学意义提供更全面的信息。