精浆piRNA对精子DNA完整性及辅助生殖技术结局的影响

目的通过检测男性不育症患者精浆中存在的睾丸特异piRNA与精子DNA损伤程度的关系,探讨其对精子DNA完整性及辅助生殖技术(ART)结局的影响。方法分析149例男性不育症患者精液质量及精子DNA完整性;按精子DNA碎片指数(DFI)将不育症患者分为:A组(DFI≤15%)、B组(15%DFI30%)、C组(DFI≥30%),采用实时荧光定量聚合酶链式反应(Q-PCR)检测各组患者精浆中piRNA-013423和piRNA-023386的表达量,比较各组患者受精率、卵裂率、优胚率、生化妊娠率及临床妊娠率的差异。结果与A、B组比较,C组的精子密度、前向活动精子、精子活动率、正常形态率均明显下降(P0.05),A、B两组比较,B组精子活动率下降(P0.05)。A、B、C三组中,C组患者piRNA-013423、piRNA-023386表达均低于A、B组(P0.01);C组患者受精率、卵裂率、优胚率、生化妊娠率及临床妊娠率均低于A、B组(P0.01)。结论男性不育症患者存在精子DNA不同程度的损伤,精浆中piRNA的水平与精子DNA完整性有关,DNA完整性差的患者精浆piRNA表达降低,且影响ART临床结局。提示piRNA可能通过某些信号通路调控精子DNA损伤,但具体机制需要下一步深入研究探讨。

Effect of semen piRNA on sperm DNA integrity and outcome of assisted reproductive technology

Objective： To explore the relationship between semen piRNA and degree of damage of sperm DNA in infertile patients by measuring the testis special piRNA, and investigate the effect of semen piRNA on sperm DNA integrity and outcome of assisted reproductive technolozy（ART）.Method： The sperm DNA integrity and semen analysis were detected in the 149 infertile patients. Then the patients were divided into three groups according to their sperm DNA fragmentation index（DFI） ; group A （ DFI ≤ 15 % ）, group B （ 15 % （ DFI （ 30 %） and group C （DFI ≥ 30 %）. Real time fluorescent quantitative polymerase chain reaction（RT-PCR）was used to detect the expression of the piRNA-013423, piRNA-023386 in semen. The rates of fertilization and cleavage, high-quality embryo, biochemical pregnancy and clinical pregnancy were compared among the three groups. Results： Compared with group A and group B, sperm concentration, activity, percentage of forward motile sperm and normal morphology sperm were significantly decreased in the group C（P〈0. 05）. The expressions of piRNA-013423 and piRNA-023386 were significantly decreased in group C（P〈0.01）. The rates of fertilization and cleavage, high-quality embryo, biochemical laregnancv and clinical pregnancy were also signKicantly decreased in group C（P〈0. 01）. Conclusions. The sperm DNA with different degree of damage is related to sperm quality in the infertile patients. The semen piRNA expression is decreased in the patients whose DNA integrity is poor. The rates of fertilization and cleavage, high-quality embryo, biochemical pregnancy and clinical pregnancy are all affected. The data of our study suggest that piRNA may regulate sperm DNA damage through some signaling pathway,and the level of piRNA can be used as one of the markers for clinical diagnosis,but the specific mechanisms need further study.