非编码小RNA在不同DNA碎片化指数的精子细胞中的表达差异

Differential expression of non-coding small RNA in spermatozoa with different DNA fragmentation index

目的探究不同精子DNA碎片化指数(DNA fragmentation index,DFI)精液精子细胞中的非编码小RNA(small non-coding RNA,sncRNAs)表达量差异。方法筛选2021年10月至2022年8月期间于海军军医大学附属长海医院生殖医学中心门诊就诊的不育男性患者纳入研究。按DFI分为DFI正常组(DFI<15%)48例、DFI临界状态组(DFI为15%~30%)40例和DFI升高组(DFI>30%)48例,比较3组患者的精子细胞中sncRNAs相对表达量的差异,并分析差异sncRNAs与精子活力的相关性。结果3组样本的年龄、精子浓度差异均无统计学意义(均P>0.05)。3组间精子活力差异有统计学意义(P<0.001),其中DFI升高组的精子活力[(30.36±4.75)%]较DFI正常组[(63.38±9.56)%,P<0.001]和DFI临界状态组[(56.50±5.87)%,P=0.034]下降。3组样本的sncRNAs靶点存在差异性表达,与其余两组相比,DFI升高组中Glu-CTC-40-10和SeC-TCA-37-4相对表达量降低,Gly-GCC、iMet-CAT-18-18和hsa-miR-151a-5p相对表达量增加,差异均有统计学意义(均P<0.001)。Spearman相关性分析显示,Glu-CTC-40-10、SeC-TCA-37-4与精子活力之间呈正相关(r=0.384,P<0.001;r=0.441,P<0.001),Gly-GCC、iMet-CAT-18-18和hsa-miR-151a-5p与精子活力之间呈负相关(r=-0.437,P<0.001;r=-0.423,P<0.001;r=-0.515,P<0.001)。结论与DFI正常的精子细胞相比,sncRNAs在DFI临界状态及升高的精子细胞中存在差异性表达,可能与DFI形成的分子机制有关,具有作为男性不育生物学标志物的潜力。

Objective To investigate the expression differences of small non-coding RNAs(sncRNAs)in sperm cells with different DNA fragmentation index(DFI).Methods Male patients who visited the Center of Reproductive Medicine of the Changhai Hospital Affiliated to Naval Medical University from October 2021 to August 2022 were included in the study.According to the DFI value,they were divided into<15%,15%-30%and>30%,which were denoted as normal DFI group(n=48),critical DFI group(n=40)and increased DFI group(n=48).The relative expression levels of sncRNAs in sperm cells of 3 groups were compared.The correlation between differential sncRNAs and sperm motility was analyzed.Results There were no significant differences in age and sperm concentration among the three groups(all P>0.05).The sperm motility in the increased DFI group[(30.36±4.75)%]was lower than that in the normal DFI group[(63.38±9.56)%,P<0.001]and the critical DFI group[(56.50±5.87)%,P=0.034].Compared with the other two groups,the relative expression levels of Glu-CTC-40-10 and SeC-TCA-37-4 in the increased DFI group decreased,while the relative expression levels of Gly-GCC,iMet-CAT-18-18,and hsa-miR-151a-5p increased,with statistical significances(all P<0.001).Spearman correlation analysis showed that Glu-CTC-40-10,SeC-TCA-37-4 were positively correlated with sperm motility(r=0.384,P<0.001;r=0.441,P<0.001),and Gly-GCC,iMet-CAT-18-18 and hsa-miR-151a-5p were negatively correlated with sperm motility(r=-0.437,P<0.001;r=-0.423,P<0.001;r=-0.515,P<0.001).Conclusion Compared with semen with normal DFI,sncRNAs are differentially expressed in semen with critical and elevated DFI,which may be related to the molecular mechanism of DFI formation.SncRNAs have the potential to be used as biological markers of male infertility.