基于串联质谱标签技术结合IPA生物信息学平台的弱精子症患者精子蛋白质组学研究

Analysis of sperm proteomics of asthenospermia patients based on tandem mass spectrometry label combined with IPA bioinformatics platform

目的应用串联质谱标签(TMT)标记技术结合IPA生物信息分析平台开展弱精子症患者精子蛋白质组学研究,为进一步研究弱精子症发病机制提供新的思路和依据。方法随机选取30例男性健康体检者和弱精子症患者的精子标本,通过TMT技术进行蛋白质组学分析。以P<0.05且表达倍数>1.7或<0.588为标准,构建候选差异蛋白质体系,利用IPA生物信息学分析平台对候选差异蛋白质进行生物信息学分析。结果通过蛋白质组学技术获得89种弱精子症患者精子候选差异蛋白质,73种蛋白质表达上调,16种蛋白质表达下调。通过IPA分析发现弱精子症患者精子候选差异蛋白质中有4种蛋白质与凋亡相关,2种蛋白质与自噬相关,1种蛋白质与雄激素相关,10种蛋白质与免疫相关,2种蛋白质与炎症有关。差异表达的蛋白质参与的主要生物学过程为蛋白质的内质网定位和核转录的mRNA分解等过程,涉及的典型信号通路为Sirtuins、蛋白激酶A和mTOR信号通路等,并获取4种与弱精子症疾病相关的候选标志物。结论弱精子症的发生与凋亡、自噬、雄激素异常、免疫及炎症等改变密切相关,但需在验证的基础上进一步研究,同时弱精子症患者精子差异蛋白质涉及复杂的生物学过程、分子功能和信号通路,为开展其发病机制研究奠定了基础。

Objective To investigate the sperm proteomics of asthenospermia patients using the tandem mass tag(TMT)labeling technique and ingenuity pathway analysis(IPA)platform for better understanding the pathogenesis of asthenospermia.Methods Sperm samples were collected from 30 male healthy subjects and 30 patients with asthenospermia and their proteomics were comparatively analyzed by TMT technology.Differentially expressed proteins were identified based on the criteria of fold change>1.7 or<0.588 and P value<0.05.The IPA bioinformatics analysis platform was employed to analyze all the differential proteins.Results A total of 89 differentially expressed proteins were obtained,including 73 upregulated proteins and 16 downregulated proteins.IPA analysis revealed that 4 differential proteins were associated with apoptosis,2 differential proteins with autophagy,1 differential protein with androgens,10 differential proteins with immune responses and 2 differential proteins with inflammation.The differentially expressed proteins were primarily involved in the endoplasmic reticulum localization of proteins and mRNA breakdown of nuclear transcription.The typical signaling pathways involved were the Sirtuins,protein kinase A,and mTOR signaling pathway.Four candidate disease markers related to asthenospermia were obtained.Conclusions The occurrence of asthenospermia is closely related to changes in apoptosis,autophagy,androgen abnormality,immunity and inflammation,but it needs to be further verified.Meanwhile,the differential proteins of sperm in patients with asthenospermia involve complex biological processes,molecular functions and signaling pathways,which lays a foundation for the study of its pathogenesis.