少精子症患者精子鱼精蛋白表达及染色质包装质量

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摘要目的：探讨鱼精蛋白（PRM1和PRM2）和精子染色质包装质量对少精子症的影响。方法：参照WHO标准收集了28例少精子症、16例畸形精子症和15例正常对照精液标本。通过提取标本总RNA，经反转录和荧光定量PCR反应分别检测PRM1、PRM2和TRF2 mRNA的相对表达量；标本纯化后进行涂片、CMA3染色，通过计算CMA3阳性率检测精子染色质包装质量。结果：少精子症组PRM1和PRM2 mRNA相对表达量分别为正常对照组的25%（P＜0.05）和20%（P＜0.05），畸形精子症组与正常对照组比较差异无统计学意义；TRF2 mRNA表达水平3组间差异无统计学意义；少精子症组CMA3阳性率（20.47%±1.19%）是正常对照组（10.69%±1.45%）的191%（P＜0.001），畸形精子症组为14.31%±2.01%，与正常对照组比差异无统计学意义。结论：鱼精蛋白（PRM1和PRM2）mRNA表达量减少可能影响精子染色质包装，使得精子形成过程异常，从而导致成熟精子数量减少。

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	洪丹
	楼哲丰
	郑九嘉
	黄学锋
	金龙金

关键词 ：男性不育, 少精子症, 精子形成, 鱼精蛋白, 染色质

Abstract：Objective: To investigate the correlation between mRNA transcription level of protamines (PRM1 and PRM2) in oligozoospermia. Methods: Sperm samples were recruited following WHO criteria and further collected from oligozoospermia patients (28 cases), teratozoospermia (16 cases) and healthy volunteers (15 cases) respectively. The mRNA transcriptions of PRM1, PRM2 and TRF2 in sperm were detected by RT-qPCR. The quantification of sperm chromatin packaging quality was performed by CMA3 staining. Results: Compared with control group, mRNA transcription levels of PRM1 and PRM2 in oligozoospermia group were significantly decreased 4 folds and 5 folds respectively. However, there’s no significant difference in mRNA transcription of PRM1 and PRM2 between teratozoospermia and control groups, or TRF2 between oligozoospermia and control groups, or TRF2 between teratozoospermia and control groups. Elvated CMA3 positive cells were only observed in oligozoospermia samples, compared with teratozoospermia and control samples. Conclusion: PRM1 and PRM2 are critical for maintaining high level of sperm counts. Decreased expression of PRM1 and PRM2 may cause sperm chromatin packaging dysfunction, which can further suppress spermiogenesis.

Key words： infertility, male oligospermia spermiogenesis protamines chromatin

收稿日期: 2017-11-02

基金资助:洪丹（1989-），女，浙江温州人，助理实验师，硕士。

作者简介: 金龙金，教授，Email：1304071636@qq.com。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2018/V48/I4/271

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