Ding Xiao-fang, Shang Xue-jun, Li Hong-gang, Guan Huang-tao, Xiong Cheng-liang
Family Planning Research Institute, Tongji Medical College, 430030, China.
Zhonghua Nan Ke Xue. 2007 May;13(5):391-5.
To study the mechanism of uPA improving sperm capacitation by investigating the effect of uPA on the mitochondrial function of mouse capacitated-sperm in vitro.
Mitochondrial function of mouse capacitated-spermatozoa was evaluated through the assessment of mitochondrial membrane potential using JC-1 performed by flow cytometer and fluorescent microscope respectively. The experiment and the control groups were designed according to the presence or absence of uPA, each divided into 5 subgroups based on the different time of uPA treatment (or BWW in the control groups) at 0, 5, 15, 30 and 60 min respectively.
(1) Compared with that at 0 min, the mean fluorescence intensity of JC-1 within the spermatozoal body and the percentage of orange-red colored spermatozoa in the experiment group were increased significantly at 5 and 15 min respectively after uPA incubation (P < 0.05). (2) The mean fluorescence intensity of JC-1 within the spermatozoal body at 15, 30 and 60 min and the percentage of orange-red colored spermatozoa at 5 and 15 min in the group were significantly higher than those in the control group (P < 0.05).
uPA could increase the mitochondrial membrane potential of mouse capacitated-spermatozoa in vitro, and maintain it at a high level for a certain period of time. By enhancing sperm mitochondrial function, uPA may provide sufficient energy for capacitated-spermatozoa to increase their motility and change their motor pattern, which might be one of the therapeutic mechanisms of uPA on male infertility.
通过研究尿激酶型纤溶酶原激活剂(uPA)对体外培养的小鼠获能精子线粒体功能的影响,探讨uPA改善精子获能的机制。
分别采用流式细胞仪和荧光显微镜,通过评估JC-1检测的线粒体膜电位来评价小鼠获能精子的线粒体功能。根据是否存在uPA设计实验组和对照组,每组再根据uPA处理(对照组为BWW)的不同时间(0、5、15、30和60分钟)分为5个亚组。
(1)与0分钟时相比,uPA孵育后实验组精子体内JC-1的平均荧光强度在5分钟和15分钟时分别显著增加,橙色精子的百分比也显著增加(P<0.05)。(2)该组在15、30和60分钟时精子体内JC-1的平均荧光强度以及在5和15分钟时橙色精子的百分比均显著高于对照组(P<0.05)。
uPA可在体外增加小鼠获能精子的线粒体膜电位,并在一定时间内维持在较高水平。通过增强精子线粒体功能,uPA可能为获能精子提供足够能量,以增加其活力并改变其运动模式,这可能是uPA治疗男性不育症的机制之一。