论文部分内容阅读
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 compare the reactivity of free Ca2 + ([Ca2 +] i) to progesterone (P) in sperm cells of normal and unexplained infertile men. Methods: Ten male patients with unexplained infertility and 9 normal fertile men were enrolled in this study. The viable spermatozoa were isolated by the upstream method and were allowed to function in vitro for 2 h. Fluorescence probes of Fluorescent probe with 8.85μmol / 3 / AM incubated in the dark 40 min load, the load of sperm suspension and the same amount of 20% gelatin mix to brake sperm, and then under the laser scanning confocal microscope dynamic observation of a sperm [Ca2] i base Level and the effect of 10μmol / LP stimulation on [Ca2 +] i. Results: The basal level of [Ca2 +] i in sperm was significantly lower in infertile group than that in reproductive group (P <0.01). Stimulation of sperm in fertile group resulted in rapid and transient increase of [Ca2 +] i and peak of [Ca2 +] i after stimulation with 10μmol / L, which was significantly higher than that of basal level (P <0.05) There was only a slight increase in [Ca2 +] i, but there was no significant difference between the peak level and the basal level (P> 0.05). The sperm [Ca2 +] i peak stimulated by P and the increase of [Ca2 +] i (difference between peak level and basal level) in fertility group were significantly higher than those in infertile group (P <0.01). CONCLUSIONS: The decreased reactivity of sperm [Ca2 +] i to P in patients with unexplained infertility suggests that there may be dysfunction of non-genotype progesterone receptors responsible for Ca2 + influx in the sperm membrane of these patients.