【摘要】 目的:观察Rho特异性的GDP解离抑制因子α(RhoGDIα)在人睾丸、精子中的表达及定位并比较RhoGDIα在正常生育男性和体外受精(IVF)不育患者精子中的表达差异。方法:通过免疫组化方法观察RhoGDIα在人睾丸中的定位;通过免疫荧光方法观察RhoGDIα在人精子获能前、获能后、顶体反应后的定位;收集正常男性精液标本(10例),高受精率(≥60%,12例)和低受精率(<60%,13例)的IVF不育患者的精液标本,Percoll细胞分离液分离精液标本,排除生精细胞和白细胞,分别通过免疫荧光和Western印迹方法,检测RhoGDIα的表达。结果:免疫组化结果显示RhoGDIα存在于人睾丸各级生精细胞中,并在长形精子细胞高表达。免疫荧光结果显示RhoGDIα在人精子的顶体和尾部有较强表达,并且随着获能的发生,在顶体上的表达减弱,当顶体反应发生后,顶体上的表达完全消失。Western印迹结果显示不易受精的IVF患者精子中RhoGDIα的表达(0.66±0.18)显著低于正常组(1.13±0.21)和易受精的IVF患者组(0.97±0.17)。结论:RhoGDIα定位于人精子的顶体和尾部,可能参与了精子运动,获能及顶体反应过程。RhoGDIα在受精率低下患者精子中表达显著降低,提示RhoGDIα可能成为一个新的男性不育的诊断指标,并且可能成为IVF供精选择的一个指标。更多还原

【Abstract】 <Objective:To investigate the expression of RhoGDIα in human testes and spermatozoa,and compare the expression of RhoGDIα in the ejaculated spermatozoa from normozoospermic me and infertilen patients men receiving in vitro fertilization(IVF).Methods: The localization of RhoGDIα in the human testis was determined by immunohistochemistry,and that in the pre-capacitated,capacitated and acrosome-reacted sperm by immunofluorescence.Western blot was used to detect the expression of RhoGDIα in the semen samples obtained from normozoospermic males(n = 10),IVF patients with high fertilization rates(≥60%,n = 12) and those with low fertilization rates(<60%,n = 13).Results: Immunohistochemistry showed that the RhoGDIα protein was located in all spermatogenic cells and highly expressed in the elongated spermatids.Immunofluorescence exhibited a high expression of RhoGDIα in the acrosome and flagellum of human sperm,which decreased in the acrosome after capacitation and disappeared after acrosome reaction.Western blot revealed an obviously decreased expression of RhoGDIα in the spermatozoa of the IVF patients with low fertilization rates(0.66 ± 0.18),with statistically significant difference from those with high fertilization rates(0.97 ± 0.17) and the normozoospermic men(1.13 ± 0.21).Conclusion: The RhoGDIα protein is located in the acrosome and flagellum of human sperm,and might be involved in sperm movement,capacitation and acrosome reaction.The significantly reduced expression of RhoGDIα in the sperm of low-fertilization patients suggests that it may be a new diagnostic biomarker for male infertility,and has a potential application value in sperm selection for IVF.更多还原