基于iTRAQ技术筛选人精子细胞生精阻滞相关候选蛋白

Screening of candidate proteins related to maturation arrest based on iTRAQ technique

目的使用同位素标记相对和绝对定量(iTRAQ)技术筛选人精子细胞生精阻滞相关候选蛋白。方法选取2015年3月至2016年9月就诊于上海交通大学附属第一人民医院男科的梗阻性无精子症及精子阻滞患者的睾丸组织各3例,进行蛋白质组学测序,对测序结果进行蛋白鉴定和分析。结果共鉴定到125种差异表达的蛋白质,21个上调,104个下调。生物学过程分析显示差异表达蛋白参与生殖过程、发育过程等。参与生殖过程占差异蛋白的29.00%。参与发育过程占差异蛋白的41.00%。细胞组分分析显示有68.00%的差异蛋白附着于细胞器上。分子功能分析显示参与绑定功能占差异蛋白的85.00%;参与转运、酶活性调节、抗氧化的蛋白比例分别为11.05%、4.62%、8.34%。参与精子变形的信号通路:如MAPK通路、Ca2+/钱调素等。结论本研究筛选到人精子细胞生精阻滞相关候选蛋白及相关信号通路,其结构和功能的异常会导致精子细胞生精阻滞,进而导致严重的男性不育症。

Objective To screen candidate proteins related to maturation arrest of humanspermatogenesis. Methods From March 2015 to September 2016, 3 cases of obstructive azoospermia and 3cases of patients with maturation arrest, who were admitted to the Shanghai General Hospital, were selectedfor proteomic sequencing, and the sequencing results were identified and analyzed. Results A total of 125differentially expressed proteins were identified, of which 21 were up-regulated and 104 weredown-regulated. Analysis of biological process shows that differentially expressed proteins participate inreproductive and developmental processes. Participation in reproductive processes accounts for 29.00% ofthe difference proteins. Participation in the development process accounted for 41.00 °/c of the differenceproteins. Cell component(CC) analysis showed that 68.00% different proteins were attached to organelles.Molecular function analysis showed that the binding function accounted for 85.00% of the differentialproteins. The proportion of proteins involved in transportation, regulation of enzyme activity andanti-oxidation were 11.05%, 4.62% and 8.34% respectively. Signal pathways involved in sperm deformationincluded MAPK pathway, Ca2+/calmodulin pathway. Conclusions The candidate proteins and itsassociated signaling pathways were selected. The abnormal structuration and function of mature arrest couldlead to severe male infertility.