雌激素对人精子的激活作用及其机制的初步研究

发布时间：2019-04-26 08:42

【摘要】：精子进入雌性生殖道后,必须经过获能、顶体反应等一系列复杂的结构和功能的改变,即被激活后才具备受精能力。精子的激活受到许多生物因素的调节,目前有关精子激活的分子机制及信号途径远未得到阐明。雌性生殖道环境中存在一定量的雌激素,尤以卵泡液及宫颈粘液中雌激素的含量最高。有研究结果表明一些动物和人精子膜上可能存在雌激素受体或雌激素结合位点,因此,精子在穿越雌性生殖道时,雌性生殖道中的雌激素可能与精子相互作用,通过特定的信号途径诱导精子激活。但是,雌激素对哺乳动物精子激活作用的普遍性还有待证明,雌激素对人精子的激活作用及其相关的信号机制也值得进一步探索。为此,我们拟采用三色法顶体染色、流式细胞术、免疫印迹技术进行以下研究:第一,用不同浓度的17β-雌二醇(E_2)分别作用生育力正常男性和不明原因不育症男性获能或非获能精子,检测精子顶体反应率(AR率)、精子内游离钙离子浓度([Ca~(2+)]_i)的变化,从而研究雌激素对人精子的激活作用及该作用对精子功能的意义。第二,比较游离雌二醇与非透膜性的雌激素-牛血清蛋白复合物(E_2-BSA)对人精子AR率、[Ca~(2+)]_i、蛋白质酪氨酸磷酸化的影响的差异;并以无钙液代替有钙液再检测E_2、E_2-BSA所致精子[Ca~(2+)]_i的变化,从而初步探讨雌激素对人精子激活的可能机制。实验结果如下:
[Abstract]:After sperm enters the female reproductive tract, it must undergo a series of complex structural and functional changes, such as capacitation, acrosome reaction and so on, that is to say, the sperm is activated before it can be fertilized. Sperm activation is regulated by many biological factors. The molecular mechanism and signal pathway of sperm activation are far from elucidated. There is a certain amount of estrogen in female reproductive tract environment, especially in follicular fluid and cervical mucus. Studies have shown that estrogen receptor or estrogen binding sites may exist in some animal and human sperm membranes. Therefore, estrogen in the female reproductive tract may interact with spermatozoa when sperm passes through the female reproductive tract. Sperm activation is induced by specific signaling pathways. However, the universality of estrogen on mammalian sperm activation has yet to be proved. The activation of estrogen on human spermatozoa and its related signal mechanisms are also worth further exploration. To this end, we plan to use trichromatic acrosome staining, flow cytometry, immunoblotting techniques to carry out the following studies: first, The sperm acrosome reaction rate (AR rate) was detected by using different concentrations of 17 尾-estradiol (E 尾-estradiol) to affect capacitated or non-capacitated spermatozoa of normal fertility men and unexplained infertile men, respectively. The changes of intracellular free calcium concentration ([Ca~ (2)] I) in spermatozoa were used to study the activation of estrogen on human spermatozoa and the significance of this effect on sperm function. Secondly, the effects of free estradiol and non-membranous estrogen-bovine serum protein complex (E_2-BSA) on human sperm AR rate, [Ca~ (2)] _ I, protein tyrosine phosphorylation were compared. The changes of [Ca~ (2)] _ I induced by E _ (2) O _ (2) and E _ (2) O-BSA in human spermatozoa were detected with calcium-free solution instead of calcium-free solution, so as to explore the possible mechanism of estrogen-induced activation of human spermatozoa. The experimental results are as follows:
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R33

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