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目的:比较外周致密纤维1(ODF1)在健康男性和弱精子症患者精子中的表达差异。方法:根据WHO标准,收集正常男性和弱精子症患者的精液标本各20份,Percoll非连续梯度离心法分离精子,收集95%Percoll以下和57%与76%Percoll层之间的精子,以排除生精细胞和白细胞的污染;采用逆转录-多聚酶链式反应(RT-PCR)和Western印迹方法,从mRNA和蛋白水平检测ODF1的表达。结果:RT-PCR结果表明,与正常男性组相比,ODF1在弱精子症患者精子中的mRNA表达水平显著降低(2.79±0.28vs1.35±0.25,P<0.05);免疫印迹与RT-PCR结果一致,与正常男性组相比,弱精子症患者精子中的ODF1蛋白的表达亦显著降低(3.64±0.34vs1.44±0.26,P<0.05)。结论:ODF1在弱精子症患者精子中的表达显著降低,提示其可能与精子活动力低下有关。
OBJECTIVE: To compare the differences in the expression of ODF1 in sperm between healthy men and asthenospermia patients. METHODS: Twenty semen samples of normal male and asthenospermia patients were collected according to the WHO criteria. Percoll non-continuous gradient centrifugation was used to separate sperm and collect sperm below 95% Percoll and between 57% and 76% Percoll layers to exclude Spermatogenic cells and leukocytes. The expression of ODF1 was detected by RT-PCR and Western blotting at mRNA and protein levels. Results: RT-PCR results showed that ODF1 mRNA expression was significantly decreased in sperms of asthenospermia patients (2.79 ± 0.28 vs 1.35 ± 0.25, P <0.05) compared with normal males. Immunoblotting and RT-PCR The results showed that the ODF1 protein expression in spermatozoa of asthenospermia patients was significantly lower than that in normal men (3.64 ± 0.34 vs 1.44 ± 0.26, P <0.05). Conclusion: The expression of ODF1 in sperms of patients with asthenospermia significantly decreased, suggesting that it may be related to the low sperm motility.