体外受精患者精子 miRNA-21，miRNA-34c，miRNA-140，miRNA-375 与胚胎质量的相关性研究

目的：通过检测不同胚胎质量的体外受精(in vitro fertilization，IVF)患者精子miRNA-21，miRNA-34c， miRNA-140，miRNA-375的表达情况，探讨精子来源的miRNAs与胚胎质量的相关性。方法：选择2012年9月至12月在 中南大学湘雅医院生殖医学中心行IVF治疗原发不育的男性患者44例，收集 IVF取卵当日剩余新鲜精液标本，采用实 时荧光定量PCR测定精子miRNAs(miRNA-21，miRNA-34c，miRNA-140，miRNA-375)的表达水平。观察胚胎情况后分 组，第3天胚胎评分的平均值<8分为实验组，≥8分为对照组。比较实验组和对照组患者一般情况及实验室资料，分 析精子miRNAs表达水平与胚胎质量的相关性。结果：实验组精子miRNA-21，miRNA-34c，miRNA-140，miRNA-375 表达水平低于对照组(P<0.01)。实验组与对照组的获卵数、减数分裂II期(metaphase II，MII)卵子数、双原核(dual pronuclear，2PN)受精数、卵裂数、受精率差异无统计学意义(P>0.05)；实验组卵裂率低于对照组(P<0.05)。精子 miRNA-21，miRNA-34c，miRNA-140，miRNA-375表达水平与第2，3天胚胎碎片率呈负相关，与第3天胚胎卵裂球数呈 正相关，与胚胎评分呈正相关。结论：精子miRNA-21，miRNA-34c，miRNA-140，miRNA-375的表达水平上升可能影 响卵裂期胚胎质量，对胚胎发育可能起一定的积极作用。

Correlation between the levels of miR-21, miR-34c,miR-140 and miR-375 in the sperm from in vitro fertilization patients and the embryo quality

Objective: To explore the correlation between sperm-originated miRNAs and embryo quality by detecting the expression levels of miR-21, miR-34c, miR-140 and miR-375 in the sperm from invitro fertilization (IVF) patients. Methods: The fresh semen specimens were collected from 44 male patients who received the IVF cycle in the Xiangya Hospital Reproductive Center from September to December in 2012. The expression levels of miR-34c, miR-140, miR-21 and miR-375 were detected by real-time fluorescent quantitative PCR. The embryos on day 2 and day 3 after fertilization were divided into the experimental and the control group, with an average embryo scores less or greater than 8, respectively. Then we compared the general and experimental data between the 2 groups respectively and analyzed the correlation between the miRNAs levels in sperm and embryo quality. Results: The expression of miR-34c, miR-140, miR-21 and miR-375 in sperms from the experimental group was significantly lower than that in the control group (P<0.05). There was no significant difference in retrieved follicles, metaphase II stage ovocytes, fertilized oocytes, cleavage number and fertilization rate between the experimental group and the control group (P>0.05), while the cleavage rate on day 2 in the experimental group was significantly lower than that of the control group (P<0.05). There was a negative correlation between the expression levels of miRNAs (miR-21, miR-34c, miR-140 and miR-375) and the ratio of fragment on day 2 or day 3. The expression levels of miR-21, miR-34c, miR-140 and miR-375 was positively correlated with the embryo score and the blastomere quantity on day 3, respectively. Conclusion: The up-regulated levels of miR-21, miR-34c, miR-140 and miR-375 in sperm may function as positive regulators in the development of cleavage stage in embryo and thus influence embryonic quantity.