弱精子症患者精子中DKKL1的表达 |
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引用本文: | 颜秋霞,马义,陈润强,周秀琴,乔静,冼英杰,冯玲,陈彩蓉. 弱精子症患者精子中DKKL1的表达[J]. 南方医科大学学报, 2018, 38(3): 324 |
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作者姓名: | 颜秋霞 马义 陈润强 周秀琴 乔静 冼英杰 冯玲 陈彩蓉 |
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摘 要: | 目的探讨精子顶体相关基因(DKKL1)在弱精子症患者精子中的表达特征及临床意义。方法收集正常男性和弱精子症患者精液,各取10 μL精液用全自动计算机辅助精液分析系统(CASA)对精液进行常规分析。为了避免白细胞和生精细胞对结果的影响,剩余精液采用4个梯度的Percoll法离心分离和纯化精子。Trizol法提取精子RNA,实时荧光定量PCR从基因水平检测DKKL1基因在正常人和弱精子症患者精子中的表达差异;最后采用Western blot方法从蛋白水平检测DKKL1蛋白在正常人和弱精子症患者精子中的表达差异。结果精液常规分析结果显示,弱精子症和正常人两组之间的年龄、精液体积、精子浓度和正常精子形态比例均没有统计学差异(P>0.05)。与正常对照组相比,弱精子症组PR 和PR+NP 精子的比例显著降低(P<0.01)。实时荧光定量PCR结果显示,与正常对照组相比,DKKL1 mRNA在弱精子症患者精子中的表达量降低11.1倍。此外,Western blot结果显示,弱精子症组DKKL1蛋白的表达量与正常对照组相比,降低了2.4倍,差异具有统计学意义(P<0.01),该结果与实时荧光定量PCR结果一致。结论DKKL1在弱精子症患者精子中的表达水平明显下降,表明其与精子运动密切相关,是导致弱精子症发生的原因之一。
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Expression of DKKL1 in spermatozoa of men with asthenospermia |
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Abstract: | Objective To compare the expression of DKKL1 in ejaculated spermatozoa of normal fertile men and men withasthenospermia and investigate the role of DKKL1 in the pathogenesis of asthenospermia. Methods The characteristics ofsemen samples collected from normal fertile men and men with asthenospermia were analyzed using computer-assistedsperm analysis according to WHO criteria. The ejaculated sperms were isolated by Percoll discontinuous density gradients todetect the expression of DKKL1 mRNA and protein using real-time PCR and Western blotting. Results The expression ofDKKL1 mRNA was significantly down-regulated by 11.1 times in asthenospermic men as compared with that in normal fertilemen (P<0.01). Western blotting showed that the expression of DKKL1 protein was down-regulated by 2.4 times inasthenospermic men compared to normal fertile men. Conclusion The expression of DKKL1, which may play an importantrole in sperm motility,is significantly decreased in ejaculated spermatozoa of men with asthenospermia. |
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