优选和冻融对精子顶体反应及顶体反应释放的顶体酶活性影响

Effects of Sperm Selection Technique and Post-thaw on Acrosome Reaction and Released Acrosin Activity of Human Semen

目的:研究优选和冻融对人精子顶体反应(acrosome reaction,AR)和AR释放的顶体酶活性影响。方法:10例健康生育男子精液,用Percoll密度梯度分离法和上游法优选精子;用3种冷冻保护剂进行冷冻;用孕酮诱导AR;采用FITC-PSA荧光染色法分析AR;采用分光光度比色法测定顶体酶活性。结果:Percoll法和上游法优选后,顶体反应率和AR释放的顶体酶活性均较优选前显著升高(P<0·05),但Percoll法和上游法优选组间比较差异不显著(P>0·05)。精子冻融后甘油组和GYC组顶体反应率和AR释放的顶体酶活性均下降,与冷冻前比较差异均有显著性(P<0·01),GYCG组与冻融前组比较,差异不显著(P>0·05)。结论:优选后顶体反应率及AR释放的顶体酶活性增加,GYCG对精子顶体反应和AR释放的顶体酶活性有保护作用。

Objective： To study the effects of selection technique and post - thaw on AR and released acrosin activity of human semen. Methods： Human semen was obtained from 10 fertile donors with normal semen parameters. Specimens were allowed to liquefy at 37℃ . Percoll - gradient treatment and swimming selection were used. Specimens were made four subgroups, before frozen and adding three different cryoprotectants （GYC; GYCG and glycerol） . Progesterone was used for induction of acrosome reaction. Acrosome reaction was analyzed using FITC - PSA staining. Acrosin activity was analyzed using spectrocolorimeter. Results： After selection, the percentage of acrosome reacted sperm and released acrosin activity were significantly higher than before （ P 〈 0. 05 ）, whereas there were no statistics differences （ P 〉 0. 05 ） between Percoll selection and swimming selection. In GYC and glycerol subgroup, the percentage of aerosome reacted sperm and released acrosin activity were both lower than that of before frozen subgroup （ P 〈 0. 01 ） . In GYCG subgroup, the percentage of acrosome reacted sperm and released acrosin activity were no difference in comparison with before frozen subgroup （ P 〉 0. 05 ） . Conclusion： Sperm selection technique increase the percentage of acrosome reacted sperm and released acrosin activity. GYCG seems to be a preferable cryoprotectant with acrosome reaction and released acrosin activity.