Henkel Ralf, Hajimohammad Marjam, Stalf Thomas, Hoogendijk Christiaan, Mehnert Claas, Menkveld Roelof, Gips Holger, Schill Wolf-Bernhard, Kruger Thinus F
Justus Liebig University and Institute for Reproductive Medicine, Giessen, Germany.
Fertil Steril. 2004 Apr;81(4):965-72. doi: 10.1016/j.fertnstert.2003.09.044.
To investigate sperm DNA damage in relation to fertilization and pregnancy.
Prospective study.
The Institute of Reproductive Medicine, Giessen, Germany.
PATIENT(S): Semen collected from 249 patients attending the IVF program.
MAIN OUTCOME MEASURE(S): The percentage of terminal deoxynucleotidyl transferase-mediated dUDP nick-end labeling- (TUNEL-), Fas-, and annexin-V-positive sperm and the proportion of green-fluorescing sperm in the acridine orange stain was determined and correlated with sperm concentration, motility, fertilization, and pregnancy.
RESULT(S): Significant correlations with the concentration of motile sperm were only found for the acridine orange stain (before and after sperm separation) and for the TUNEL assay (after sperm separation). Moreover, patients whose sperm had a high percentage of DNA fragmentations showed significantly lower pregnancy rates (TUNEL assay: 19.05% vs. 34.65%; acridine orange stain: 24.58% vs. 37.93%). The apoptosis parameters (annexin V binding and Fas expression) showed no statistically significant differences.
Our data clearly demonstrate that DNA fragmentation, as determined by the TUNEL assay, is predictive for pregnancy in IVF. This implies that spermatozoa with DNA fragmentation can still fertilize an oocyte but that when paternal genes are "switched on," further embryonic development stops, resulting in failed pregnancy. It seems that, at least in the patients we analyzed, apoptosis in the sperm does not play a role for fertilization. This would imply that DNA fragmentation in human spermatozoa is caused by external factors, such as reactive oxygen species, rather than by apoptosis.
研究精子DNA损伤与受精及妊娠的关系。
前瞻性研究。
德国吉森生殖医学研究所。
从249名接受体外受精项目的患者收集精液。
测定末端脱氧核苷酸转移酶介导的dUDP缺口末端标记法（TUNEL）、Fas和膜联蛋白V阳性精子的百分比以及吖啶橙染色中发绿色荧光精子的比例，并将其与精子浓度、活力、受精及妊娠情况进行关联分析。
仅在吖啶橙染色（精子分离前后）和TUNEL检测（精子分离后）中发现与活动精子浓度存在显著相关性。此外，精子DNA碎片化百分比高的患者妊娠率显著较低（TUNEL检测：19.05%对34.65%；吖啶橙染色：24.58%对37.93%）。凋亡参数（膜联蛋白V结合和Fas表达）无统计学显著差异。
我们的数据清楚地表明，通过TUNEL检测确定的DNA碎片化可预测体外受精中的妊娠情况。这意味着有DNA碎片化的精子仍能使卵母细胞受精，但当父本基因“开启”时，进一步的胚胎发育停止，导致妊娠失败。似乎至少在我们分析的患者中，精子凋亡在受精过程中不起作用。这意味着人类精子中的DNA碎片化是由外部因素如活性氧引起的，而非凋亡所致。
