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[明胶底物玻片法简化顶体酶检测方法的开发及临床应用研究]

[Studies for development and clinical use of the simplified method of acrosin measurement by using gelatin substrate slides].

作者信息

Hasegawa T

出版信息

Nihon Sanka Fujinka Gakkai Zasshi. 1985 Dec;37(12):2713-21.

PMID:3910739
Abstract

A simplified method for the detection of acrosin proteolytic activity (APA) of the individual sperm was developed by using a gelatin substrate slide. The following results were obtained in clinical application. Soybean trypsin inhibitor and TLCK completely inhibited APA at concentrations of 500 micrograms/ml and 1.5 mg/ml, respectedly. APA in diluted semen increased with greater dilution reaching the maximum at 8 to 10 fold dilution. Even at this dilution, APA was only 70% of the APA measured with washed sperm. A significant correlation was found between APA and the sperm concentration in male infertility. APA increased as the concentration of sperm increased, and the lowest APA was observed at less than 20 X 10(6)/ml. An unexplained male infertility group had a significantly lower APA than a fertile group. The group with both more than 40% motility and 2b quality had a higher APA than other groups with poor motility. There was no significant correlation between the rate of penetration of the zona-free hamster ova seen in the penetration assay and APA. Sperm-immobilizing antibody had no effect on APA. Nonoxynol-9 (N-9), a spermicidal agent, could inhibit APA completely at concentrations of 250 micrograms/ml and more. A linear regression curve of APA was obtained with a log-dose concentration of N-9. From these results it is felt that this newly developed method will be useful in clinical practice.

摘要

通过使用明胶底物载玻片,开发了一种用于检测单个精子顶体蛋白酶活性(APA)的简化方法。在临床应用中获得了以下结果。大豆胰蛋白酶抑制剂和甲苯磺酰-L-赖氨酸氯甲基酮分别在浓度为500微克/毫升和1.5毫克/毫升时完全抑制APA。稀释精液中的APA随着稀释倍数的增加而增加,在8至10倍稀释时达到最大值。即使在这种稀释度下,APA也仅为用洗涤过的精子测得的APA的70%。在男性不育症中,发现APA与精子浓度之间存在显著相关性。APA随着精子浓度的增加而增加,在精子浓度低于20×10⁶/毫升时观察到最低的APA。不明原因的男性不育组的APA明显低于生育组。活力超过40%且质量为2b级的组的APA高于其他活力差的组。在穿透试验中观察到的无透明带仓鼠卵穿透率与APA之间没有显著相关性。精子固定抗体对APA没有影响。杀精剂壬苯醇醚-9(N-9)在浓度为250微克/毫升及更高时可完全抑制APA。获得了N-9的对数剂量浓度与APA的线性回归曲线。从这些结果来看,认为这种新开发的方法将在临床实践中有用。

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