Abstract：
Many cellular processes in spermatozoa, including apoptosis and motility, are regulated by miRNA. Different miRNAs and molecular pathways are involved in asthenozoospermia (AS) conditions, which are thought to be one of the causes of infertility with reduced sperm motility. Thirty-two semen samples from four Holstein bulls with normozoospermia (NS), total motility ≥ 70%, and progressive motility ≥ 60%, and 32 semen samples from four bulls with AS, total motility ≤ 40%, and progressive motility ≤ 32% were used to investigate the function of apoptosis-related miRNAs in the AS group. Samples were then aspirated into a 0.5 mL straw after dilution with a Tris-egg yolk extender and frozen at -196°C. After freezing, semen samples were thawed for 2 weeks at 37°C and sperm kinematic parameters, plasma membrane integrity, acrosome integrity, DNA fragmentation, apoptosis status, and expression of apoptosis-related miRNAs (miR-2114, miR-296-3p, miR-455-3p, and miR345-3p) were evaluated. Our results showed that the functional and flow cytometric parameters of the NS group were significantly better than those of the AS group. In the NS group, miR-455-3pp and miR-2412 were upregulated, while miR-345-3p was downregulated compared with the AS group. In the AS group, miR-296-39, miR-2412, and miR-345-3p levels were strongly correlated with membrane integrity, DNA fragmentation, and apoptosis status. The findings demonstrated that the selected miRNAs based on bioinformatic analysis in AS and NS samples had a substantial association with functional and flow cytometry indicators and may be involved in regulating apoptosis and motility in AS samples.
摘要：
精子的许多细胞过程，包括细胞凋亡和运动性，受miRNA调控。不同的miRNA和分子通路参与弱精子症（AS）条件，这被认为是精子活力下降的不孕症的原因之一。来自四只正常精子症（NS）的荷斯坦公牛的32个精液样本，总活动力≥70%，和渐进运动≥60%，和四头公牛的32个精液样本，总活动力≤40%，研究AS组细胞凋亡相关miRNAs的功能。然后用Tris-蛋黄增量剂稀释后将样品吸入0.5mL吸管中，并在-196°C下冷冻。冷冻后,精液样本在37°C解冻2周，精子运动学参数，质膜完整性，顶体完整性,DNA片段化,凋亡状态，和凋亡相关miRNA的表达（miR-2114，miR-296-3p，miR-455-3p,和miR345-3p）进行了评估。我们的结果表明，NS组的功能和流式细胞仪参数明显优于AS组。在NS组中，miR-455-3pp和miR-2412上调，而miR-345-3p与AS组相比下调。在AS组中，miR-296-39、miR-2412和miR-345-3p水平与膜完整性密切相关。DNA片段化,和凋亡状态。研究结果表明，在AS和NS样本中基于生物信息学分析选择的miRNA与功能和流式细胞术指标具有实质性关联，并且可能参与AS样本中的凋亡和运动调节。
