用改良的底物玻片法检测TLCK对人精子顶体蛋白酶活性的影响

Modified Substrate Slide Test in Determination of Effect of TLCK on Human Sperrn Acrosin Proteolytic Activity

精子顶体蛋白酶是精子在女性生殖道内运行及穿透卵透明带与卵子结合而完成受精的特异性主酶之一。检测精子顶体蛋白酶活性(APA)对正确评价精子的受精能力及探讨顶体酶抑制剂的机理,筛选及开发新型避孕药都有明确的意义。本文介绍经本实验室改良的明胶底物玻片法检测人精子顶体蛋白酶活性,以精子顶体溶解明胶基质所显示的平均光轮直径(MHD)作为 APA 的指标,测得正常功能精子 APA 的 MHD 为29.96μm。证实该法能最大限度地显示 APA的光轮直径,操作简便、实用,可有效地用于临检。同时以该法测到顶体酶抑制剂 TLCK 的浓度梯度效应,验证了该法的可靠性。

Sperm acrosin proteolytic activity(APA)is one kind of enzyme for sperm move-ment in the female reproductive tract and penetration through the zona pellucida.The measurement of sperm APA is very important for testing the insemination abi-lity of sperm and investigating the effect and mechanism of inhibitors of acrosinenzyme.It is also useful for selecting and screening of new contraceptives.A modi-fide gelatin substrate slide test for measuring human sperm APA is reported.A halocould be seen on the gelatin with the dissolution of the sperm acrosin.The meanhalo diameter(MHD)was used as an index for the measurement of APA.We foundthe MHD of normal sperm APA was 29.96 μm,This proved that in the method thehalo diameter of APA could significantly be shown,hence it was an easy processand was very useful for clinical tests.This method was also used to test the effectof acrosin inhibitor,TLCK.