Wip1基因在人精子中的表达及与精子浓度和活力的相关性研究

摘要

目的:研究野生型p53诱导的磷酸酶1(Wip1)基因在人类精子中的表达情况及其与精子浓度及活力的相关性.方法:收集正常对照组、少精子症组、弱精子症组和少弱精子症组精液样本,每组20例.检测4组精子DNA碎片指数(DFI)、精子存活率.分别采用实时荧光定量PCR(qPCR)法和免疫荧光技术检测精子Wip1基因和蛋白的表达.结果:少精子症组、弱精子症组、少弱精子症组DFI均较正常对照组显著升高(均P<0.05).少精子症组与正常对照组精子存活率无明显差异(P>0.05),弱精子症组、少弱精子症组较正常对照组显著降低(P<0.05).DFI≥25％组精子存活率和活力均较DFI<25％组显著降低(P<0.05),而两组精子浓度无明显差异(P>0.05).正常对照组与少精子症组Wip1基因和蛋白表达水平均无明显差异(P>0.05),弱精子症组、少弱精子症组Wip1基因和蛋白表达水平均较正常对照组显著升高(均P<0.05).Pearson相关分析显示:Wip1基因表达与精子浓度和DFI呈正相关关系,与精子活力呈负相关关系(均P<0.05).结论:Wip1在人类精子中有表达,且其可能负调控精子DNA损伤修复进程,Wip1表达上调可导致精子活力降低,DFI升高,并影响精子浓度.%Objective:To investigate the expression of wild-type p53-induced phosphatase 1(Wip1) gene in human sperm and to explore its correlation with sperm concentration and motility.Methods:Semen samples of normal semen,oligozoospermia,weak sperm disease and oligospermia were collected,20cases in each group.The sperm DNA fragmentation index(DFI) and sperm survival rate were detected.The expression of Wip1was detected by real-time fluorescent quantitative PCR(qPCR) and immunofluorescence.Results:The DFI of oligozoospermia,weak sperm disease and oligospermia groups was significantly higher than that of normal control group(P<0.05).There was no significant difference in sperm survival rate between the oligozoospermia group and the normal group(P>0.05) ,the survival rate of the weak spermatozoa group and the oligospermia group were significantly lower than that of the normal group(P<0.05).The difference between survival rate and viability between DFI>25％ and DFI≤25％ groups was statistically significant(P<0.05) ,but there was no significant difference in sperm concentration between the two groups(P>0.05).There was no significant difference in Wip1mRNA expression between the normal group and the oligozoospermia group(P>0.05) ,and the weak spermatozoa group and the oligospermia group had higher Wip1mRNA level compared with the normal group(P<0.05).Pearson correlation analysis showed that Wip1expression was negatively associated with sperm motility,while positively correlated with sperm concentration and DFI(P<0.05).Conclusion: Wip1expressed in human sperm,and it might negatively modulate sperm DNA damage.Up-regulation of Wip1expression could decrease sperm motility,increase DFI and affect sperm concentration.