苯甲基磺酰氟在低温保存人类精子中的应用

Analysis of the influence of PMSF on human sperm in low temperature preservation

目的:探讨苯甲基磺酰氟(Phenylmethanesulfonyl fluoride,PMSF)对低温保存的人类精子存活率及DNA双链不完整性的影响。方法:将新采集的80份人类精液分成两等份,其中一份加入PMSF冷冻保护剂,另一份不加入PMSF冷冻保护剂,将两份标本保存在4℃冰箱中,通过实验比较精子在两种状态下30min^120h内精子活率及DNA双链不完整率差异。结果:低温保存30min^120h,不加入冷冻保护剂组精子平均活率及DNA双链不完整率分别为:(73.2±2.74)%、(67.3±2.56)%、(56.5±2.42)%、(47.4±2.18)%、(41±2.58)%、(30.8±2.64)%;(7.3±1.35)%、(13.8±1.26)%、(16.3±1.56)%、(25±1.36)%、(34.2±1.34)%、(39.2±1.52)%;低温保存30min^120h,加入冷冻保护剂组精子平均活率及DNA双链不完整率分别为:(73.2±2.36)%、(70.2±2.68)%、(59.8±2.46)%、(51.2±2.68)%、(44.5±2.95)%、(33±2.89)%;(7.3±1.45)%、(11.5±1.64)%、(14.3±1.68)%、(22±1.58)%、(30.8±1.65)%、(35.7±1.87)%。结论:低温保存30min^120h之内,精子活率及DNA双链不完整率的差异有统计学意义,在同样的条件下加入PMSF冷冻保护剂同不加入PMSF冷冻保护剂进行低温保存相比,精子的平均活率和DNA双链不完整率的差异无统计学意义。使用与不使用PMSF冷冻保护剂对人类精子进行低温保存都是可行的,如使用PMSF,最佳保护效果只维持在48h之内,之后随时间延长,精子活率降低,DNA双链不完整率升高,120h之后精子几乎没有活率。

To investigate the effect of PMSF on the alive-rate and integrity of DNA double chain in cryopreservation.Methods：The fresh human semen was divided into two groups with PMSF cryoprotectant in one group.Semen in both groups were stored in refrigerators of 4℃,to compare the alive-rate and integrity of DNA double chain after Cryopreservation method.Results：（30m-120h in cryopreservation,using free-cryopr-eservation）The average alive-rate and non-integrity of DNA double chain of the human semen in experimental group were：（73.2 ±2.74）%,（67.3 ±2.56）%,（56.5 ±2.42）%,（47.4 ±2.18）%,（41 ±2.58）%, （30.8 ±2.64）%;（7.3 ±1.35）%,（13.8 ±1.26）%,（16.3 ±1.56）%,（25 ±1.36）%,（34.2 ±1.34）%,（39.2 ±1.52）%;（30m-120h in Cryopreservation method,using PMSF）.The average alive-rate and non-integrity of DNA double chain of the human semen in experimental group were：（73.2 ±2.36 ）%,（70.2 ± 2.68）%,（59.8 ±2.46）%,（51.2 ±2.68）%、（44.5 ±2.95）%,（33 ±2.89）%;（7.3 ±1.45）%,（11.5 ± 1.64）%,（14.3 ±1.68）%,（22 ±1.58）%,（30.8 ±1.65）%,（35.7 ±1.87）%.Conclusion：The differences of alive-rate and integrity of DNA double chain of the human semen are of statistic significance.PMSF is optional in the cryopreservation of human semen.When using the PMSF,the best protective effect only lasts for 48 hours, and as time goes on,the alive-rate will decrease while the incidence of non-integrity of DNA double chain goes up,with no alive-rate after 120 hours.