【作者】 陆培信；

【导师】 张家颖；

【作者基本信息】 吉林大学 ， 免疫学， 2007， 硕士

【摘要】 近年来,男性不育的患病率逐渐上升,对男性不育的研究越来越引起人们的关注。男性不育症的发生一个重要原因是精子异常,研究精子中蛋白C抑制剂(PCI)基因的多态性,探讨蛋白C抑制剂基因多态性是否与男性精子的生成障碍相关。在研究中,首先在临床收集精液标本,在精子中提取基因组DNA,根据人类基因组数据库设计特异引物,进行PCR扩增,琼脂糖凝胶电泳鉴定,将PCR产物回收纯化进行TA克隆测序验证,将测序结果与GenBank数据库比对分析。本实验旨在探讨精子中蛋白C抑制剂基因SNP G10877T位点的多态性分布,研究蛋白C抑制剂(PCI)基因SNP位点的基因型和等位基因频率与男性不育相关性,以探讨男性不育的原因,为男性不育的基因诊断和合理治疗提供科学依据。更多还原

【Abstract】 More and more attention was put into the study of male infertility with the growing frequency. The percent of sperm abnormality caused by inheritance is 30%. The molecular aetiology of male subfertility is still unknown in the majority of cases and it is thought that multiple genes are involved. It is important to study the presence of gene polymorphism in the protein C inhibitor (PCI) gene.Protein C inhibitor is a plasma glycoprotein belonging to the serpin superfamily of serine protease inhibitors. PCI acts as an inhibitor of activated protein C, but also inhibits various other proteases such as thrombin, plasma kallikrein, tissue kallikrein, urokinase, acrosin and prostate-specific antigen (PSA). PCI is expressed in many organs and tissues and is present in various human body fluids. The highest PCI concentration has been described in seminal plasma. PCI is also present in the seminal vesicles, the testis and the prostate. It is improved that PCI gene is one of genes that might play a role in male reproductive function. The presence of PCI gene is necessary for male reproduction.Purpose: We detected the presence of point mutation in PCI gene from sperm of male subfertility, then performed a mutation screen of the PCI gene in subfertile men to study the connection of polymorphism PCI gene with male subfertility, we builded a quickly, highly special diagnosis method, produced a basis for the treatment. On the other hand, we have also builed the technique of screening of PCI gene mutation to help the reproduction.Method: Sperm was obtained from 53 cases normal male and 102 cases male subfertility, genome DNA of sperm was extracted. Complete sequence analyses were performed according to gene bank online. We performed PCR by special PCR primers, sequenced the amplification, then compared with data ingene bank to perform the statistical analyses.Result: There are statistical significant differences between the TT and allele genotype frequencies of male subfertility group and control group after the statistical analysis. The frequency of T allele in male subfertility group is significantly higher then that of control group.Conclusion: The polymorphism of PCI gene is connects with male subfertility in the North-east of China.There are variability of gene mutation in different races and fields of different groups, it is necessary to indicate further the relationship between the polymorphism of PCI gene and the number of abnormal sperms in male subfertility. There is importance to supply the basis theory of the inheritance, diagnosis prevention and treatment in male subfertility.更多还原