Division of Gynecological Endocrinology and Reproductive Medicine, Department of Gynecology and Obstetrics, Ludwig-Maximilians-University (LMU) Munich, Campus Grosshadern, Marchioninistrasse 15, 81377 Munich, Germany.
Clinical Division of Gynecologic Endocrinology and Reproductive Medicine, Department of Obstetrics and Gynecology, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria.
Hum Reprod. 2017 Aug 1;32(8):1574-1582. doi: 10.1093/humrep/dex224.
Are unexplained recurrent miscarriages associated with abnormal protamine-1 and protamine-2 mRNA levels in spermatozoa?
Both protamine-1 and protamine-2 mRNA levels as well as the protamine-1 to protamine-2 mRNA ratio in spermatozoa from men whose female partners experienced two or more consecutive miscarriages were significantly different compared to those from both healthy control men and subfertile couples undergoing IVF/ICSI.
Aberrant sperm protamine ratios are known to be associated with male-factor infertility. Data from this study suggest that the protamine mRNA ratio may additionally affect early embryo development.
STUDY DESIGN, SIZE, DURATION: The study population was recruited from men whose female partners presented with two or more consecutive unexplained miscarriages in a consultation for recurrent pregnancy loss between 2014 and 2016. At the research laboratory of the Urological Clinic of the University Giessen, spermatozoa from cases and controls were subjected to reverse transcription quantitative PCR (RTqPCR) using specific primer pairs for protamine-1 and protamine-2.
PARTICIPANTS/MATERIALS, SETTING, METHODS: Protamine-1 and protamine-2 mRNA levels were analysed in semen samples from 25 men whose female partners experienced at least two consecutive idiopathic miscarriages before the 20th week of gestation. The couples were recruited during consultation at the Fertility Center of the LMU Munich, Germany, and at the Clinical Division of Gynecologic Endocrinology and Reproductive Medicine of the Medical University of Vienna, Austria. Results were compared with those from 32 healthy donors (WHO, 2010) recruited at the Department of Urology, Pediatric Urology and Andrology, Giessen, Germany, and 107 men whose partners participated in an IVF/ICSI program at the Fertility Center of the LMU Munich, Germany.
Protamine-1 and protamine-2 mRNA levels as well as the protamine mRNA ratio and all routine semen parameters revealed significant differences between recurrent miscarriage couples and healthy volunteers (P < 0.01). When comparing recurrent miscarriage couples with couples undergoing IVF/ICSI, Ct-values of protamine-1 and protamine-2 mRNAs were significantly higher and the protamine mRNA ratio was significantly lower in RM couples (P < 0.01). When comparing protamine mRNA levels and the protamine mRNA ratio with routine semen parameters, a significant negative correlation was evident between progressive motility and the protamine-2 mRNA level (P = 0.015), as well as between non-progressive motility and the protamine mRNA ratio (P = 0.023).
Although our data demonstrate significant abnormalities in RM, larger sample sizes will be needed to confirm our results. Larger sample sizes should also balance the fact that we had to focus mainly on median protamine mRNA levels. Finally, men in the healthy control group were younger in age than those in the case group, which might have introduced some bias, at least concerning the classic semen parameters. Moreover, only protamine mRNA instead of protein levels could be measured.
Although the exact mechanism remains to be elucidated, our data suggest that protamine mRNA levels in spermatozoa are not only important for successful fertilization, but also for proper development of the early embryo.
STUDY FUNDING/COMPETING INTEREST(S): Grant from the University Clinic Giessen and Marburg (UKGM 29/2015GI). There are no competing interests.
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不明原因的反复性流产是否与精子中异常的鱼精蛋白-1 和鱼精蛋白-2 mRNA 水平有关？
与健康对照组男性和接受 IVF/ICSI 的亚生育夫妇相比，经历两次或两次以上连续流产的男性伴侣的精子中鱼精蛋白-1 和鱼精蛋白-2 mRNA 水平以及鱼精蛋白-1 与鱼精蛋白-2 mRNA 比值均显著不同。
已知异常的精子鱼精蛋白比值与男性因素不育有关。本研究的数据表明，鱼精蛋白 mRNA 比值可能还会影响早期胚胎发育。
研究设计、大小和持续时间：该研究人群是从 2014 年至 2016 年期间因反复妊娠丢失而就诊的连续不明原因流产两次或两次以上的男性伴侣中招募的。在吉森大学泌尿科诊所的研究实验室中，使用特定的引物对针对鱼精蛋白-1 和鱼精蛋白-2 的逆转录定量 PCR（RTqPCR）对病例和对照组的精子进行分析。
参与者/材料、设置、方法：分析了 25 名男性的精液样本中的鱼精蛋白-1 和鱼精蛋白-2 mRNA 水平，这些男性的女性伴侣在妊娠 20 周前至少经历了两次连续的特发性流产。这些夫妇是在德国慕尼黑 LMU 生育中心和奥地利维也纳医科大学妇科内分泌学和生殖医学临床分部的咨询中招募的。结果与在德国吉森泌尿科、小儿泌尿科和男科系招募的 32 名健康供体（世界卫生组织，2010 年）和在德国慕尼黑 LMU 生育中心参加 IVF/ICSI 计划的 107 名男性进行了比较。
反复性流产夫妇与健康志愿者之间的鱼精蛋白-1 和鱼精蛋白-2 mRNA 水平以及鱼精蛋白 mRNA 比值和所有常规精液参数均显示出显著差异（P <0.01）。当将反复性流产夫妇与接受 IVF/ICSI 的夫妇进行比较时，RM 夫妇的鱼精蛋白-1 和鱼精蛋白-2 mRNAs 的 Ct 值显着更高，鱼精蛋白 mRNA 比值显着更低（P <0.01）。当将鱼精蛋白 mRNA 水平和鱼精蛋白 mRNA 比值与常规精液参数进行比较时，在渐进性运动和鱼精蛋白-2 mRNA 水平之间（P=0.015）以及在非渐进性运动和鱼精蛋白 mRNA 比值之间（P=0.023）存在显着的负相关。
局限性/谨慎原因：尽管我们的数据表明 RM 存在显着异常，但需要更大的样本量来证实我们的结果。更大的样本量还应平衡我们主要关注中位数鱼精蛋白 mRNA 水平的事实。最后，健康对照组中的男性比病例组中的男性年龄更小，这可能会引入一些偏见，至少在经典精液参数方面是这样。此外，只能测量精子中的鱼精蛋白 mRNA 水平，而不是蛋白质水平。
虽然确切的机制仍有待阐明，但我们的数据表明，精子中的鱼精蛋白 mRNA 水平不仅对成功受精很重要，而且对早期胚胎的正常发育也很重要。
研究资金/利益冲突：来自吉森和马尔堡大学诊所（UKGM 29/2015GI）的资助。没有利益冲突。
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