Cryopreservation-induced human sperm DNA damage is predominantly mediated by oxidative stress rather than apoptosis

被引:344
|
作者
Thomson, L. K. [1 ]
Fleming, S. D. [2 ]
Aitken, R. J. [3 ,4 ]
De Iuliis, G. N. [3 ,4 ]
Zieschang, J. -A. [1 ]
Clark, A. M. [1 ]
机构
[1] Fertil First, Hurstville, NSW 2220, Australia
[2] Univ Sydney, Westmead Hosp, Dept Obstet & Gynaecol, Westmead, NSW 2000, Australia
[3] Univ Newcastle, Discipline Biol Sci, Callaghan, NSW 2308, Australia
[4] Univ Newcastle, ARC Ctr Excellence Biotechnol & Dev, Callaghan, NSW 2308, Australia
关键词
Sperm DNA fragmentation; cryopreservation; 8OHdG; genistein; caspase inhibitor; DEOXYRIBONUCLEIC-ACID FRAGMENTATION; SPONTANEOUS LIPID-PEROXIDATION; HUMAN-SPERMATOZOA; IN-VITRO; EJACULATED SPERMATOZOA; HYDROGEN-PEROXIDE; INTEGRITY; GENISTEIN; CASPASES; FERTILIZATION;
D O I
10.1093/humrep/dep214
中图分类号
R71 [妇产科学];
学科分类号
100211 ;
摘要
Whereas studies have revealed that the cryopreservation of human semen increases sperm DNA fragmentation, the mechanisms involved in this type of cryo-injury are largely unknown. Elucidation of these mechanisms may provide insight into preventing such injury. We obtained 60 semen samples from 60 men and conducted experiments to determine the cause of cryopreservation-induced DNA fragmentation using 8-oxo-7,8-dihydro-2'deoxyguanosine (8OHdG) as a biomarker of oxidative stress, percentage caspase positive cells as an indicator of apoptosis, the potential antioxidant genistein and the caspase inhibitor Z-VAD(OMe)-FMK. Cryopreservation led to a significant increase in percentage DNA fragmentation, percentage 8OHdG and percentage caspase positive cells (P < 0.001). Percentage DNA fragmentation was positively correlated with percentage 8OHdG before (r = 0.756, P < 0.001) and after cryopreservation (r = 0.528, P = 0.017). The addition of 50 and 100 mu M genistein to the cryoprotectant had a significant protective effect on sperm DNA (P < 0.001) although the caspase inhibitor demonstrated no difference to the control. Human sperm DNA fragmentation is associated with an increase in oxidative stress during cryopreservation, rather than the activation of caspases and apoptosis. The estrogenic compound genistein may be useful in reducing this effect but larger trials are needed to confirm this.
引用
收藏
页码:2061 / 2070
页数:10
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