孕酮对正常生育和不明原因不育男性精子细胞内游离Ca^(2+)浓度的影响  

Effects of Progesterone on Intracellular Free Ca^(2+) Concentration in the Spermatozoa of Fertile Men and Patients with Unexplained Infertility

作　　者：霍涌玮[1] 王丽蓉[1] 田宏[1] 许拥军 赵晓鸽[3] 邱曙东[1] 

机构地区：[1]西安交通大学医学院生殖医学研究中心,陕西西安710061 [2]新安中心医院外科,陕西西安710048 [3]西安交通大学医学院生物医学研究实验中心,陕西西安710061

出　　处：《中华男科学杂志》2009年第11期980-984,共5页National Journal of Andrology

基　　金：陕西省自然科学研究计划项目(2004C234)~~

摘　　要：目的:比较正常生育男性和不明原因不育男性患者精子细胞内游离Ca2+浓度([Ca2+]i)对孕酮(P)的反应性。方法:选择10例不明原因不育男性患者和9例正常生育男性,用上游法分离活力良好的精子并于体外获能2 h,将获能后精子与8.85μmol/L钙荧光探针Fluo-3/AM避光孵育40 min负载,将负载后的精子悬液与等量20%明胶混匀以制动精子,然后在激光扫描共聚焦显微镜下动态观察单个精子头部的[Ca2+]i基础水平以及10μmol/L P刺激对[Ca2+]i的影响。结果:不育组获能2 h后精子的[Ca2+]i基础水平显著低于生育组(P<0.01)。生育组精子经10μmol/L P刺激后,产生快速而短暂的[Ca2+]i升高,[Ca2+]i峰值水平显著高于其基础水平(P<0.05);而不育组精子经P刺激后,[Ca2+]i仅有微弱增加,其峰值水平与基础水平相比无统计学差异(P>0.05)。生育组由P激发的精子[Ca2+]i峰值水平以及[Ca2+]i升高幅度(峰值水平与基础水平之差)均显著高于不育组(P<0.01)。结论:不明原因不育男性患者精子[Ca2+]i对P的反应性降低,提示这些患者精子膜上负责Ca2+内流的非基因型孕激素受体可能存在功能障碍。Objective： To investigate the difference in the responsiveness of intracellular free Ca2＋ concentration（i） to progesterone in the spermatozoa of normal fertile men and patients with unexplained infertility.Methods： Nine normal fertile men and 10 patients with unexplained infertility were selected in this study.After swim-up separation of the motile fraction and 2-hour in vitro capacitation,the spermatozoa were loaded with the fluorescent calcium indicator Fluo-3/AM（8.85 μmol/L） for 40 minutes away from the light,and then the sperm suspension was mixed with equal amount of 20% gelatin to immobilize the spermatozoa.The basal intracellular free i and that induced by 10 μmol/L progesterone in the individual sperm were assessed by laser scanning confocal microscopy.Results： The infertile patients had a significantly lower basal level of i in the capacitated sperm than the fertile men（P 〈 0.01）.The sperm from the normal controls responded to progesterone by exhibiting a rapid but transient rise in i,with the peak level significantly higher than the basal level（P 〈 0.05）,while those from the infertile patients by showing a slight increase,with no significant difference between the peak and basal levels（P 〉 0.05）.Both the peak of the progesterone-induced i and its increase amplitude expressed as the difference between the peak and basal levels were significantly higher in the normal fertile group than in the infertile patients（P 〈 0.01）.Conclusion： The responsiveness of i to progesterone is reduced in the spermatozoa of patients with unexplained infertility,which suggests a functional defect in the non-genomic sperm membrane progesterone receptor responsible for calcium influx.

关 键 词：精子 男性不育 孕酮 钙离子 顶体反应 

分 类 号：R698.2[医药卫生—泌尿科学]