Amjadian Tayebeh, Yaghmaei Parichehreh, Nasim Hayati Roodbari, Yari Kheirollah
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Heliyon. 2023 Oct 16;9(10):e21099. doi: 10.1016/j.heliyon.2023.e21099. eCollection 2023 Oct.
Male infertility accounts for nearly 40%-50% of all infertile cases. One of the most prevalent disorders detected in infertile men is errors in the MEST differentially methylated region (DMR), which has been correlated with poor sperm indexes. The aim of our study was to characterize the methylation pattern of the MEST gene, along with assessing seminal factors and chromatin condensation in sperm samples from both infertile patients and fertile cases, all of whom were candidates for intracytoplasmic sperm injection. We collected forty-five semen specimens from men undergoing routine spermiogram analysis at the Infertility Treatment Center. The specimens consisted of 15 samples of normospermia as the control group, 15 individuals of asthenospermia, and 15 individuals of oligoasthenoteratospermia as the cases group. Standard semen analysis and the chromatin quality and sperm maturity tests using aniline blue staining were performed. The DNA from spermatozoa was extracted and treated with a sodium bisulfite-based procedure. The methylation measure was done quantitatively at the DMRs of the gene by quantitative methylation-specific polymerase chain reaction (qMSP). The mean percentages of total motility, progression, and morphology in normospermia were significantly higher than oligoasthenoteratospermia and asthenospermia, and they were substantially higher in asthenospermia compared to oligoasthenoteratospermia (P ≤ 0.05). The mean percentages of histone transition abnormality and methylation in oligoasthenoteratospermia were significantly higher than asthenospermia and normospermia (P ≤ 0.05). A negative correlation existed between the histone transition abnormality and methylation with sperm parameters. In conclusion, chromatin integrity, sperm maturity, and methylation may be considered important predictors for addressing male factor infertility. Therefore, we suggest that male infertility may be linked to methylation of the imprinted genes.
男性不育症占所有不育病例的近40%-50%。在不育男性中检测到的最普遍的疾病之一是MEST差异甲基化区域（DMR）的错误，这与精子指标不佳有关。我们研究的目的是表征MEST基因的甲基化模式，并评估不育患者和可育病例精子样本中的精液因素和染色质凝聚情况，所有这些人都是胞浆内单精子注射的候选者。我们从不育治疗中心接受常规精子图分析的男性中收集了45份精液标本。标本包括作为对照组的15份正常精液样本、15例弱精子症个体和15例少弱畸精子症个体作为病例组。进行了标准精液分析以及使用苯胺蓝染色的染色质质量和精子成熟度测试。从精子中提取DNA并采用基于亚硫酸氢钠的方法进行处理。通过定量甲基化特异性聚合酶链反应（qMSP）对该基因的DMR进行定量甲基化测定。正常精液中总活力、前向运动和形态的平均百分比显著高于少弱畸精子症和弱精子症，并且弱精子症中的这些指标相比少弱畸精子症也显著更高（P≤0.05）。少弱畸精子症中组蛋白转化异常和甲基化的平均百分比显著高于弱精子症和正常精液（P≤0.05）。组蛋白转化异常和甲基化与精子参数之间存在负相关。总之，染色质完整性、精子成熟度和甲基化可能被视为解决男性因素不育的重要预测指标。因此，我们认为男性不育可能与印记基因的甲基化有关。
