抗Eppin抗体抑制人精子活性运动及相关机制

Inhibition of sperm activated motility by the anti-Eppin antibody and the potential mechanisms

目的:阐明抗附睾蛋白酶抑制剂(Eppin)抗体抑制人精子动力及分析相关机制。方法:将正常男性精子经过percoll分选后分为7组:空白对照组,抗体对照组与40μg·ml-1抗兔多克隆抗体孵育,其余5组分别与不同浓度抗Eppin抗体(浓度分别为2.5、5、10、20和40μg·ml-1)共同孵育4 h后进行精子动力测定,线粒体膜电位(Δψ)、ATP浓度及电镜镜检检测。结果:随着抗Eppin抗体浓度的增加,精子活力下降并呈浓度依赖趋势;当抗Eppin抗体浓度超过5μg·ml-1,精子Δψ及产生ATP浓度显著下降(P<0.05);部分精子线粒体正常环形结构发生异常改变,亚线粒体膜变薄甚至消失。结论:抗Eppin抗体与人精子共孵育后精子线粒体功能及超微结构异常,导致精子活性运动显著下降。

Objective：To clarify the inhibition of sperm activated motility by the anti-Eppin antibody and the potential mechanisms .Methods： Semen samples were obtained by masturbation from normal donors .The motile sperm were collected through discontinuous percoll gradient centrifugation and divided into 7 groups .The sperm in one group were incubated with the 40 μg· ml -1 anti-rabbit antibody and other five groups were incubated with the 2.5, 5, 10, 20 or 40 μg · ml-1 anti-Eppin antibody .Results： After the sperm treated with the anti-Eppin antibody , the results of computer-assisted semen analysis , mitochondrial membrane potential （Δψ） together with ATP production assays measurement and the images of electron microscopy show the reduction of the activated motility,Δψand ATP production , and the aberrations of the helical pattern of mitochondria with the partial loss of sub-mitochondrial reticulum .Conclusion： After the human sperm incubated with the anti-Eppin antibody , the aberrations in ultra structure and the functions of the mitochondria of the sperm result in the reduction of the sperm activated motility .