苯甲基磺酰氟(PMSF)在犬精子4℃保存中的作用  

Effect of Phenylmethylsulfonyl Fluoride(PMSF) on Canine Sperm Stored at 4℃

作　　者：马啸[1] 叶华虎[2] 曾林[2] 路静[1] 王迎[1] 陈振文[1] 

机构地区：[1]首都医科大学基础医学院,北京100069 [2]军事医学科学院实验动物中心,北京100071

出　　处：《中国比较医学杂志》2009年第2期46-49,F0003,共5页Chinese Journal of Comparative Medicine

基　　金：科技条件平台服务首都建设专项项目(No:Z0006303041231)

摘　　要：目的探讨4℃保存的条件下,苯甲基磺酰氟(phenylmethylsulfonyl fluoride,PMSF)对犬精子的保护作用。方法犬精子以含不同浓度PMSF的Tris-卵黄液(Tris-egg yolk,EYT)处理后,于4℃低温保存,在24、48、72、96 h分别以伊红、瑞-吉和吖啶橙染色检测精子活率、顶体膜和DNA完整性。结果低温保存24 h后,吖啶橙染色检测DNA断裂率(%):对照组(5.93±0.32),实验组:20μg/mL(4.50±0.38)、40μg/mL(4.20±0.25)、80μg/mL(4.21±0.46)、160μg/mL(3.87±0.67),P<0.05;各浓度组之间没有明显差异;72 h后,犬精子活率仍>30%;96 h内,对照组和实验组犬精子在活率、顶体膜完整性上未见明显差异。结论PMSF于4℃低温保存条件下对犬精子具有保护作用,24 h内能有效保护精子DNA双链的完整性,4℃低温保存72 h后,犬精子仍符合人工授精要求。Objective To investigate the effect of PMSF in storage of canine sperm at 4℃. Methods Canine semen was stored in Tris-egg yolk extender with or without PMSF at 4℃. After 24 h, 48 h, 72 h and 96 h, the motility of sperm, the integrity of aerosome membrane and DNA fragementation were examined by eosin staining, Wright-Giemsa staining and acridine orange staining, respectively. Results After 24 h, The DNA fragmentation indexes （DFI） （ % ） by acridine orange staining were as following： control group （5：93 ± 0.32）, 20 μg/mL experimental group （4.50 ± 0.38）, 40μg/mL experimental group （4.20 ± 0.25）, 80 g/mL experimental group （4.21 ± 0.46）, and 160 μg/mL experimental group （3.87 ± 0.67）, and there was no significant difference between any two expermental groups （ P 〈 0.05）. After 72 h, the motility of canine sperms was still above 30%. There was no significant difference between the values of sperm motility and integrity of aerosome membrane in control and experirmental groups after storage for 92 h. Conclusion PMSF can protect the DNA integrity of canine sperm at 4 ℃ within 24 h, canine sperm can still be used to artificial insemination within 72 h stored at 4℃.

关 键 词：犬精子 苯甲基磺酰氟 低温保存 DNA断裂率 

分 类 号：R-33[医药卫生]