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. 2018 Mar 20;38(3):324-328.
doi: 10.3969/j.issn.1673-4254.2018.03.13.

[Expression of DKKL1 in spermatozoa of men with asthenospermia]

[Article in Chinese]
Qiu-Xia Yan  1 , Yi Ma, Run-Qiang Chen, Xiu-Qin Zhou, Jing Qiao, Ying-Jie Xian, Ling Feng, Cai-Rong Chen
Affiliations

[Expression of DKKL1 in spermatozoa of men with asthenospermia]

[Article in Chinese]
Qiu-Xia Yan et al. Nan Fang Yi Ke Da Xue Xue Bao. .

Abstract
in English, Chinese

Objective: To compare the expression of DKKL1 in ejaculated spermatozoa of normal fertile men and men with asthenospermia and investigate the role of DKKL1 in the pathogenesis of asthenospermia.

Methods: The characteristics of semen samples collected from normal fertile men and men with asthenospermia were analyzed using computer-assisted sperm analysis according to WHO criteria. The ejaculated sperms were isolated by Percoll discontinuous density gradients to detect the expression of DKKL1 mRNA and protein using real-time PCR and Western blotting.

Results: The expression of DKKL1 mRNA was significantly down-regulated by 11.1 times in asthenospermic men as compared with that in normal fertile men (P<0.01). Western blotting showed that the expression of DKKL1 protein was down-regulated by 2.4 times in asthenospermic men compared to normal fertile men.

Conclusion: The expression of DKKL1, which may play an important role in sperm motility,is significantly decreased in ejaculated spermatozoa of men with asthenospermia.

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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Figures

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1
Western blot检测DKKL1蛋白在正常对照组和弱精子症患者精子中的表达 DKKL1 protein in semen from normal or asthenozoospermia males by Western blotting. 1-4: Asthenospermia; 5-8: Normal.
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