Center of Reproductive Medicine, Huai'an Maternity and Child Health Care Center, Huai'an, China.
Mol Reprod Dev. 2021 Feb;88(2):158-166. doi: 10.1002/mrd.23454. Epub 2021 Jan 28.
The dysregulation of microRNAs (miRNAs) plays an important role in asthenozoospermia. This study evaluated the sperm microRNA-423-5p (miR-423-5p) expression in asthenozoospermia and normozoospermia, exploring the role of miR-423-5p in asthenozoospermia. Eighty participants were divided into asthenozoospermic (AZS, n = 40) and normozoospermic (Norm, n = 40) groups. Fresh semen samples were collected and the sperm cells were separated. Quantitative Real-Time polymerase chain reaction was used to measure the sperm miR-423-5p level. Receiver operating characteristic curve (ROC) was employed to test the diagnostic performance of miR-423-5p in asthenospermia. Dual-reporter luciferase assay was adopted to confirm the target gene of miR-423-5p. The target gene level in asthenozoospermia and normozoospermia was measured, and the biological function of target gene in asthenozoospermia was evaluated. Results showed that the miR-423-5p expression level in the AZS group was higher than that in Norm group, which was positively correlated with the severity of asthenozoospermia. ROC analysis of miR-423-5p showed an area under curve (AUC) of 0.69 (95% confidence interval = 0.57-0.80, p <0 .01), with 80% sensitivity and 60% specificity. Glutathione S-transferase mu 1 (GSTM1) is a target gene of miR-423-5p, which significantly decreased in the AZS group. Compared with Norm group, glutathione S-transferase (GST) activity and total antioxidant capacity (TAC) level decreased, while malondialdehyde (MDA) level increased in the AZS group. Furthermore, GST activity and TAC level were negatively correlated with miR-423-5p expression, while MDA level was positively correlated with miR-423-5p expression. In conclusion, the sperm miR-423-5p level significantly was upregulated in asthenozoospermia. High-level miR-423-5p inhibited sperm motility through targeting GSTM1 to promote oxidative stress.
miRNAs 的失调在弱精症中起着重要作用。本研究评估了弱精症和正常精子中的精子 microRNA-423-5p（miR-423-5p）表达，探索了 miR-423-5p 在弱精症中的作用。80 名参与者被分为弱精症（AZS，n=40）和正常精子（Norm，n=40）组。收集新鲜精液样本并分离精子细胞。采用实时定量聚合酶链反应（qRT-PCR）测量精子 miR-423-5p 水平。采用受试者工作特征曲线（ROC）测试 miR-423-5p 在弱精子症中的诊断性能。采用双报告基因荧光素酶报告基因实验证实 miR-423-5p 的靶基因。测量弱精症和正常精子中的靶基因水平，并评估靶基因在弱精症中的生物学功能。结果显示，AZS 组的 miR-423-5p 表达水平高于 Norm 组，且与弱精症的严重程度呈正相关。miR-423-5p 的 ROC 分析显示曲线下面积（AUC）为 0.69（95%置信区间 0.57-0.80，p<0.01），具有 80%的敏感性和 60%的特异性。谷胱甘肽 S-转移酶 mu 1（GSTM1）是 miR-423-5p 的靶基因，在 AZS 组中明显减少。与 Norm 组相比，AZS 组的谷胱甘肽 S-转移酶（GST）活性和总抗氧化能力（TAC）水平降低，丙二醛（MDA）水平升高。此外，GST 活性和 TAC 水平与 miR-423-5p 表达呈负相关，而 MDA 水平与 miR-423-5p 表达呈正相关。总之，弱精症中精子 miR-423-5p 水平显著上调。高水平的 miR-423-5p 通过靶向 GSTM1 抑制精子运动能力，促进氧化应激。
