uPAR在维持人精子正常功能方面体外实验探究.doc

uPAR在维持人精子正常功能方面体外实验探究[摘要] 目的:探讨uPAR在维持精子活力、活率以及参与顶体反应过程中的作用。方法:将不同浓度的抗uPAR抗体与精子共同孵育,同时设立对照组,于10、30、60 min行精液常规分析。先加入获能液诱导精子获能,而后将不同浓度的抗uPAR抗体与精子共同孵育60 min,同时设立对照组,诱发精子发生顶体反应,通过流式细胞仪检测顶体的完整性。结果:随着抗uPAR抗体浓度的升高和孵育时间的延长,精子运动能力呈现不断下降趋势。高浓度的抗uPAR抗体可降低精子顶体反应率。结论:uPAR与其配体结合后可以影响精子的运动、获能和顶体反应。 [关键词] uPAR;抗uPAR抗体;活率;活力;顶体反应 [中图分类号] R392.7[文献标识码]A [文章编号]1673-7210(2009)07(a)-023-02 Experimental research of influence of uPAR on keeping human sperm normal function in vitro HAO Junjie, XU Jixiu (Department of Urology, the First Hospital of Shanxi Medical University, Taiyuan030001,China) [Abstract] Objective: To study the effect of uPAR on human sperm viability, motility rate, and acrosome reaction(AR).Methods: The spermatozoa solution was incubated with the different concentrations of anti-uPAR antibody in vitro. Measurements were carried out at 10, 30 and 60 min, including sperm motility and viability, while establishing normal control group.the spermatozoa solution was incubated with the different concentrations of anti-uPAR antibody in vitro. Acrosome reaction induced by anti-uPAR antibody was measured by flowcytometer, while establishing normal control group. Results: Sperm motility function showed a downward trend with increasing anti-uPAR antibody, concentration and incubated time. Anti-uPAR antibody of high concentration could induce AR. Conclusion: uPAR has effects on human sperm motility function, capacitation and acrosome reaction. [Key words] uPAR; Anti-uPAR antibody; Motility; Viability; Acrosome reaction 尿激酶型纤溶酶原激活因子受体(uPAR)又称CD-87,是一种新的纤溶因子。在细胞迁移、新生血管的生成、组织重建以及肿瘤细胞的侵袭转移等生理和病理过程中发挥重要作用[1]。已有实验证明,uPA同男性生殖系统关系密切,参与调解精子成熟、排放[2]、获能、活力、受精以及精液液化[3-4]等生殖生理过程。然而,作为uPA的特异性受体,uPAR在男性生殖系统方面所起作用的研究并不多。本实验的研究目的就是探讨uPAR在维持精子活力、活率以及参与精子顶体反应过程中的作用,并以此推测uPA和uPAR对维持人精子正常生理功能的意义。 1资料与方法 1.1研究对象 搜集精液常规正常的志愿供精者,按照WHO标准从中筛选20份精液样本。 1.2主要试剂 抗uPAR抗体、PBS缓冲液(武汉博士德生工)、Earle’s液、FITC-PSA(Sigma公司)、人血清清蛋白(华兰生工)、孕酮(天津金耀氨基酸有限公司)、BWW储备液、BWW培养液。 1.3方法 1.3.1