Reproductive Medical Center, Henan Province Key Laboratory for Reproduction and Genetics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China.
Reproductive Medical Center, Henan Province Key Laboratory for Reproduction and Genetics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China.
Life Sci. 2020 Sep 1;256:117895. doi: 10.1016/j.lfs.2020.117895. Epub 2020 Jun 2.
We aimed to investigate the effect of sperm miR-34c on early human embryonic development kinetics and clinical outcomes of in vitro fertilization (IVF) patients.
After oocyte insemination, residual sperm specimens were collected from 58 patients undergoing IVF. miR-34c expression levels in sperm, oocytes, zygotes, and embryos/blastocysts were detected with qRT-PCR, and embryonic development kinetics were observed using time-lapse technology. To confirm the role of miR-34c in regulation of early embryonic development, miR-34c siRNA was injected into zygotes obtained from in vitro-matured oocytes. A ROC curve was used to determine the cutoff value. Comparisons of embryonic development kinetics and clinical outcomes were performed according to the cutoff value.
The miR-34c expression level was highest in 3PN zygotes, but was not expressed in human oocytes. In the miR-34c siRNA group, embryonic development kinetic parameters t2, t3, t4, and t5 were significantly prolonged, but the cleavage rate and high-quality embryo rate were lower than in the control group. The levels of sperm miR-34c were negatively correlated with t5 and positively correlated with rates of blastocyst formation, high-quality blastocysts, and pregnancy. The miR-34c levels and the blastocyst formation rate were higher in the pregnancy group (p < 0.05). Logistic regression analysis showed that sperm miR-34c level was significantly correlated with pregnancy (OR: 5.056, 95% CI: 1.560-16.384; p = 0.007).
The sperm miR-34c expression level is associated with embryonic development kinetics and clinical outcomes. Thus, miR-34c expression is beneficial to embryonic development and may be used as an indicator of IVF outcomes.
本研究旨在探讨精子 miR-34c 对体外受精（IVF）患者早期胚胎发育动力学和临床结局的影响。
在卵母细胞受精后，收集 58 例行 IVF 患者的剩余精子标本。采用 qRT-PCR 检测精子、卵母细胞、受精卵和胚胎/囊胚中的 miR-34c 表达水平，并利用时差技术观察胚胎发育动力学。为了验证 miR-34c 在调节早期胚胎发育中的作用，将 miR-34c siRNA 注入体外成熟卵母细胞获得的受精卵中。采用 ROC 曲线确定截断值。根据截断值比较胚胎发育动力学和临床结局。
3PN 受精卵中 miR-34c 表达水平最高，但在人卵母细胞中不表达。在 miR-34c siRNA 组中，胚胎发育动力学参数 t2、t3、t4 和 t5 显著延长，而卵裂率和优质胚胎率低于对照组。精子 miR-34c 水平与 t5 呈负相关，与囊胚形成率、优质囊胚率和妊娠率呈正相关。妊娠组的 miR-34c 水平和囊胚形成率较高（p<0.05）。Logistic 回归分析显示，精子 miR-34c 水平与妊娠显著相关（OR：5.056，95%CI：1.560-16.384；p=0.007）。
精子 miR-34c 表达水平与胚胎发育动力学和临床结局相关。因此，miR-34c 的表达有利于胚胎发育，可作为 IVF 结局的指标。
