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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。
AIM: To observe the expression and localization of RhoGDIα in human testis and sperm and to compare the expression of RhoGDIα in sperm between normal fertile male and infertile in vitro fertilization (IVF) patients. Methods: Immunohistochemistry was used to observe the localization of RhoGDIα in human testis. Immunofluorescence staining was used to observe the localization of RhoGDIα in human sperm before and after capacitation and acrosome reaction. Normal male semen samples (10 cases) Semen samples of IVF infertile patients with high fertilization rate (≥60%, 12 cases) and low fertilization rate (<60%, 13 cases) were separated by Percoll cells, and spermatogenic cells and leucocytes were excluded. Fluorescence and Western blotting were used to detect the expression of RhoGDIα. Results: Immunohistochemistry showed that RhoGDIα was present in spermatogenic cells at all levels of human testis and was highly expressed in spermatogenic cells. Immunofluorescence results showed that RhoGDIα was strongly expressed in the acrosomes and tails of human spermatozoa, and as the capacitation took place, the expression on the acrosome decreased. When the acrosome reaction occurred, the acrosome expression completely disappeared. Western blotting showed that expression of RhoGDIα in spermatozoa (0.66 ± 0.18) was significantly lower in non-fertile IVF patients than in normal controls (1.13 ± 0.21) and in IVF susceptible patients (0.97 ± 0.17). Conclusion: RhoGDIα is localized in the acrosome and tail of human spermatozoa, and may be involved in sperm motility, capacitation and acrosome reaction. RhoGDIα was significantly decreased in sperm of patients with low fertilization rate, suggesting that RhoGDIα may be a new diagnostic indicator of male infertility and may be an indicator of IVF selection.