Ropporin在正常男性和弱精子症患者精液精子中的表达差异

Differential expression of Ropporin in ejaculated spermatozoa from health volunteers and the patients with asthenospermia

目的比较Ropporin在健康男性和弱精子症患者精液精子中的表达差异。方法收集健康男性和弱精子症患者的精液精子,为了排除生精细胞和白细胞的污染,精液标本经非连续性Percoll梯度离心和分离纯化。采用荧光定量RT-PCR和免疫细胞化学方法,从基因和蛋白水平检测精子Ropporin的表达。结果荧光定量RT-PCR显示,Ropporin mRNA在正常男性和弱精子症患者精液精子中均有表达。与正常男性相比, Ropporin在弱精子症患者精子中的表达显著降低。免疫细胞化学结果显示,Ropporin蛋白在正常男性精子的表达水平明显高于弱精子症患者,该结果与荧光定量RT-PCR结果相一致。结论Ropporin在弱精子症患者精子中的表达显著降低,可能是导致精子运动能力低下的原因之一。

Objective To comparatively analyze the expression of Ropporin in ejaculated spermatozoa from health volunteers and the patients with asthenospermia. Methods Semen samples were obtained from health volunteers and the patients with asthenospermia （A〈25% or A＋B〈30%） from the Center for Reproductive Medicine, Peking University Shenzhen Hospital. Semen analysis was performed according to WHO criteria. To rule out the contamination of germ cells and leucocytes, human ejaculated spermatozoa were purified by a discontinuous Percoll density gradient centrifugation. The expressions of Ropporin in spermatozoa from the two groups were detected by Real-time RT-PCR and immunocytochemistry simultaneously. Results The mRNA expression of ropporin in spermatozoa from asthenozoospermic men was significantly downregulated than that from health volunteers. To confirm the results of real-time RT-PCR, the expression of Ropporin protein in human ejaculated spermatozoa was further detected by immunoeytoehemistry. The results showed that the expression of Ropporin protein was significantly decreased in the patients with asthenospermia, Compared as that in healthy volunteers, which was similar to the results from real-time RT-PCR. Conclusion The expression of Ropporin in ejaculated spermatozoa found to be decreased significantly in the patients with asthenospermia, which might be associated with low motility of sperm.