Mirsanei Jamileh Sadat, Gholipour Hadis, Zandieh Zahra, Jahromi Masoumeh Golestan, Masroor Mojgan Javedani, Mehdizadeh Mehdi, Amjadi Fatemehsadat
Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Reproductive Sciences and Technology Research Center, Department of Anatomical Sciences, Iran University of Medical Sciences, Tehran, Iran.
Clin Exp Reprod Med. 2023 Sep;50(3):185-191. doi: 10.5653/cerm.2023.05890. Epub 2023 Aug 1.
Although intracytoplasmic sperm injection (ICSI) is a way to deal with in vitro fertilization failure, 3% of couples still experience repeated fertilization failure after attempted ICSI, despite having sperm within normal parameters. These patients are a challenging group whose sperm cannot fertilize the egg during ICSI. Unfortunately, no test can predict the risk of fertilization failure. Phospholipase C zeta (PLCζ) and transition nuclear proteins (TNPs) are essential factors for chromatin packaging during sperm maturation. This study aimed to assess PLCζ1 and TNP1 expression in the sperm of patients with fertilization failure and the correlations among the DNA fragmentation index, PLCζ1 and TNP1 gene and protein expression, and the risk of fertilization failure.
In this study, 12 infertile couples with low fertilization rates (<25%) and complete failure of fertilization in their prior ICSI cycles despite normal sperm parameters were chosen as the case group. Fifteen individuals who underwent ICSI for the first time served as the control group. After sperm analysis and DNA fragmentation assays, quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and Western blot analyses were performed to compare the gene and protein expression of PLCζ and TNP1 in both groups.
DNA fragmentation was significantly higher in the fertilization failure group. The qRT-PCR and Western blot results demonstrated significantly lower PLCζ and TNP1 gene and protein expression in these patients than in controls.
The present study showed that fertilization failure in normozoospermic men was probably due to deficient DNA packaging and expression of TNP1.
尽管卵胞浆内单精子注射(ICSI)是处理体外受精失败的一种方法,但仍有3%的夫妇在尝试ICSI后仍经历反复受精失败,尽管精子参数正常。这些患者是一个具有挑战性的群体,其精子在ICSI过程中无法使卵子受精。不幸的是,没有检测方法能够预测受精失败的风险。磷脂酶Cζ(PLCζ)和过渡核蛋白(TNPs)是精子成熟过程中染色质包装的关键因素。本研究旨在评估受精失败患者精子中PLCζ1和TNP1的表达,以及DNA碎片化指数、PLCζ1和TNP1基因及蛋白表达与受精失败风险之间的相关性。
在本研究中,选择12对受精率低(<25%)且既往ICSI周期中尽管精子参数正常但受精完全失败的不育夫妇作为病例组。15名首次接受ICSI的个体作为对照组。在进行精子分析和DNA碎片化检测后,采用定量逆转录聚合酶链反应(qRT-PCR)和蛋白质免疫印迹分析来比较两组中PLCζ和TNP1的基因和蛋白表达。
受精失败组的DNA碎片化程度显著更高。qRT-PCR和蛋白质免疫印迹结果显示,这些患者的PLCζ和TNP1基因及蛋白表达明显低于对照组。
本研究表明,正常精子男性的受精失败可能是由于DNA包装不足和TNP1表达缺陷所致。