17β雌二醇对人精子功能的非基因组调节效应及其机制的研究

Nongenomic Effects and Mechanisms of 17β-Estradiol on Motility and Fertilizing Ability in Human Sperm

目的：探讨17β雌二醇对人精子功能非基因组效应的可能机制。方法：运用计算机辅助精子分析系统与人精子穿透去透明带金黄地鼠卵异种体外受精实验,评价17β雌二醇对精子功能的调节作用；精子经信号转导途径抑制剂处理后,再采用流式细胞术检测精子胞内Ca^(2+)浓度的变化。结果：1μmol/L和5μmol/L E_2-BSA能提高人精子的A级精子百分比、平均曲线运动速度、平均直线运动速度、平均路径速度(P＜0．05)；自身受精率偏低的继发不育患者精子经1μmol/L和5μmol/L E_2-BSA作用后受精率提高(P＜0．05),但对本身受精率较高的精子影响不大(P＞0．05)；应用腺苷酸环化酶、磷脂酶C和酪氨酸蛋白激酶特异性抑制剂分别作用后均明显抑制E_2-BSA引起的胞内Ca^(2+)浓度的上升(P＜0．05)。结论：17β雌二醇能显著提高精子的运动能力和受精率,17β雌二醇可能通过腺苷酸环化酶、磷脂酶C或酪氨酸蛋白激酶信号转导途径实现对人精子功能调节的非基因组效应,为改善人精子功能提供了可能的新途径。

Objective： To study the mechanisms of nongenomic effects of estradiol in human sperms. Methods： Sperms were stimulated by impermeable 17β-estradiol（E2-BSA）, then the motility was determined by sperm formular assay and the fertilizing ability was determined by sperm penetration of zona-free hamster egg assay. The intracellular calcium（[Ca^2＋]i） in sperm was measurcd by flow cytometry after the sperm were treated with E2-BSA and the inhibitors of trans-membrane signaling transduction pathways. Results： There was no significant μmol/L E2-BSA on the motility and fertilization rate of human sperm （P〉0.05）, μmol/L and 5 μmol/L E2-BSA significantly increased the motility and fertilization rate of human spermatozoa in short time （P〈0.05）.The [Ca^2＋]i in sperm was decreased after the spermatozoa were treated with the inhibitors of adenylyl cyclase, phospholipase C or protein tyrosine kinase compared with the sperm treated with E2-BSA only. Conclusion： E2-BSA can rapidly increase the motility and fertilizing ability of human sperm and the possible trans-membrane signaling transduction pathways may include the adenylyl cyclase pathway, the phospholipase C and protein tyrosine kinase pathway.