Department of Obstetrics & Gynaecology, Saarland University, Homburg, Saar, Germany.
Department of Basic Medical Sciences, Faculty of Medicine, Yarmouk University, Irbid, 21163, Jordan.
J Assist Reprod Genet. 2021 Aug;38(8):2021-2029. doi: 10.1007/s10815-021-02199-w. Epub 2021 Apr 24.
The purpose of the present study was to determine the relationship between infertility and the polymorphisms of mitochondrial NADH dehydrogenase subunit 4 (MTND4) by spermatozoa analysis in fertile and subfertile men.
Samples were divided into 68 subfertile men (case group) and 44 fertile men (control group). After semen analysis, samples were purified. The whole genome was extracted using a QIAamp DNA Mini Kit and the mitochondrial DNA was amplified by using the REPLI-g Mitochondrial DNA Kit. Polymerase chain reaction (PCR) was used to amplify the MT-ND4 gene. Then, samples were purified and sequenced using the Sanger method.
Twenty-five single-nucleotide polymorphisms (SNPs) were identified in the MTND4 gene. The genotype frequencies of the study population showed a statistically significant association between rs2853495 G>A (Gly320Gly) and male infertility (P = 0.0351). Similarly, the allele frequency test showed that rs2853495 G>A (Gly320Gly) and rs869096886 A>G (Leu164Leu) were significantly associated with male infertility (adjusted OR = 2.616, 95% CI = 1.374-4.983, P = 0.002; adjusted OR = 2.237, 95% CI = 1.245-4.017, P = 0.007, respectively).
In conclusion, our findings suggested that male infertility was correlated with rs2853495 and rs869096886 SNPs in MTND4.
本研究旨在通过对生育力正常和生育力低下男性的精子分析，确定线粒体 NADH 脱氢酶亚单位 4（MTND4）多态性与不孕之间的关系。
将样本分为 68 名生育力低下男性（病例组）和 44 名生育力正常男性（对照组）。精液分析后，对样本进行纯化。使用 QIAamp DNA Mini Kit 提取全基因组，使用 REPLI-g Mitochondrial DNA Kit 扩增线粒体 DNA。采用聚合酶链反应（PCR）扩增 MT-ND4 基因。然后，使用 Sanger 法对样本进行纯化和测序。
在 MTND4 基因中鉴定出 25 个单核苷酸多态性（SNP）。研究人群的基因型频率显示 rs2853495 G>A（Gly320Gly）与男性不育之间存在统计学显著关联（P=0.0351）。同样，等位基因频率测试显示 rs2853495 G>A（Gly320Gly）和 rs869096886 A>G（Leu164Leu）与男性不育显著相关（调整后的 OR=2.616，95%CI=1.374-4.983，P=0.002；调整后的 OR=2.237，95%CI=1.245-4.017，P=0.007）。
总之，我们的研究结果表明，男性不育与 MTND4 中的 rs2853495 和 rs869096886 SNP 相关。
