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精子DNA碎片化是否会影响卵裂球活检后的发育潜能及细胞凋亡发生率?

Does sperm DNA fragmentation affect the developmental potential and the incidence of apoptosis following blastomere biopsy?

作者信息

Haghpanah Tahereh, Salehi Mohammad, Ghaffari Novin Marefat, Masteri Farahani Reza, Fadaei-Fathabadi Fatemeh, Dehghani-Mohammadabadi Maryam, Azimi Hadi

机构信息

a Department of Reproductive Biology and Anatomical Sciences , Faculty of Medicine .

b Department of Transgenic Animal Science , Stem Cell Technology Research Center .

出版信息

Syst Biol Reprod Med. 2016;62(1):1-10. doi: 10.3109/19396368.2015.1103324. Epub 2015 Dec 17.

Abstract

Common methods employed in assisted reproduction technology (ART) include intracytoplasmic sperm injection (ICSI) with an unspecified level of sperm DNA fragmentation (SDF) and preimplantation genetic diagnosis (PGD). The aim of this study was to investigate the impact of SDF on human preimplantation embryo development and the incidence of apoptosis following a single blastomere biopsy. Using sperm chromatin dispersion (SCD) to assess SDF, a total of 20 processed semen samples were categorized into two groups; group I: SDF ≤30% and group II: SDF >30%. After ICSI, fertilization, cleavage, and embryo quality score were assessed. A single blastomere was biopsied from day 3 embryos and development was monitored on day 4. The frequency of apoptosis in biopsied embryos was assayed by TUNEL and the level of BCL-2, BAX, hsa-mir-15a, and hsa-mir-16-1 were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). SCD was found to be negatively correlated with sperm motility and normal form spermatozoa (p < 0.05). The rate of fertilization, cleavage, and embryo quality score were not significantly different between the two groups (all p > 0.05). SDF >30% had no negative effect on potential development and did not increase the proportion of apoptotic cells and the level of apoptosis-related genes and microRNAs (miRNAs) in group II vs. group I (p > 0.05). It appears that at the levels assessed paternal genome damage had little if any negative effect on preimplantaton embryo development and apoptosis following single blastomere biopsy. This may reflect the selection of morphologically normal sperm for ICSI and the repair capacity of the oocyte.

摘要

辅助生殖技术(ART)中常用的方法包括精子DNA碎片率(SDF)未明确的胞浆内单精子注射(ICSI)和植入前基因诊断(PGD)。本研究的目的是探讨SDF对人类植入前胚胎发育以及单个卵裂球活检后细胞凋亡发生率的影响。使用精子染色质扩散(SCD)评估SDF,将总共20份处理后的精液样本分为两组;第一组:SDF≤30%,第二组:SDF>30%。ICSI后,评估受精、卵裂和胚胎质量评分。从第3天的胚胎中活检单个卵裂球,并在第4天监测其发育情况。通过TUNEL法检测活检胚胎中的细胞凋亡频率,并通过定量实时聚合酶链反应(qRT-PCR)评估BCL-2、BAX、hsa-mir-15a和hsa-mir-16-1的水平。发现SCD与精子活力和正常形态精子呈负相关(p<0.05)。两组之间的受精率、卵裂率和胚胎质量评分无显著差异(所有p>0.05)。与第一组相比,第二组中SDF>30%对潜在发育没有负面影响,也没有增加凋亡细胞的比例以及凋亡相关基因和微小RNA(miRNA)的水平(p>0.05)。看来在所评估的水平上,父本基因组损伤对单个卵裂球活检后的植入前胚胎发育和细胞凋亡几乎没有负面影响。这可能反映了ICSI时对形态正常精子的选择以及卵母细胞的修复能力。

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