Reproductive Medical Center, Guangdong Women and Children Hospital, Guangzhou, China.
Children Inherit Metabolism and Endocrine Department, Guangdong Women and Children Hospital, Guangzhou, China.
Syst Biol Reprod Med. 2021 Apr;67(2):127-136. doi: 10.1080/19396368.2020.1846822. Epub 2020 Dec 29.
In this study, we aimed to explore the potential differences in proteomic profiles from the testicular tissue of azoospermatic men with impaired spermatogenesis and normal spermatogenesis. Isobaric tags for relative and absolute quantitation (iTRAQ) labeled technology and LC-MS/MS technology were used to identify differentially expressed proteins. Potential functions of differentially expressed proteins were predicted using gene ontology (GO) and the Kyoto encyclopedia of genes and genomes (KEGG). Immunohistochemistry (IHC) and western blot (WB) were used to verify the differentially expressed proteins. A protein-protein interaction (PPI) network was built to outline the regulatory network of differentially expressed proteins. A total of 3,945 proteins were identified in men with normal and impaired spermatogenesis. Of these, 116 proteins were differentially expressed in men with impaired spermatogenesis: 39 were upregulated and 77 were downregulated. Furthermore, we found that these differentially expressed proteins were mainly involved in the cellular component, which may be mainly associated with the spliceosome, ribosome, and thyroid hormone synthesis signaling pathways. The spliceosome- and ribosome-associated proteins YBX1, FBL, and HNRNPU were downregulated. And the proteomic profile of testicular tissue in men with impaired spermatogenesis is different from that of men with normal spermatogenesis. For this reason, differentially expressed proteins such as YBX1, FBL and HNRNPU might be involved in the pathology of spermatogenesis dysfunction. iTRAQ: Isobaric tags for relative and absolute quantitation;GO: Gene ontology; KEGG: Kyoto encyclopedia of genes and genomes; IHC: Immunohistochemistry; WB: Western blot; PPI: Protein-protein interaction; ICSI: Intracytoplasmic sperm injection; BP: Biological process; CC: Cellular components; MF: Molecular function; snoRNA: Small nucleolar RNA; snRNA: Small nuclear RNA; LC-MS/MS: Liquid chromatography and MS/MS analysis; BSA: Bovine serum albumin; SD: Spermatogenic dysfunction; micro-TESE: Testicular microscopic sperm extraction.
在这项研究中,我们旨在探索生精障碍和正常精子发生的无精子症男性睾丸组织中蛋白质组图谱的潜在差异。使用同位素相对标记与绝对定量(iTRAQ)标记技术和 LC-MS/MS 技术鉴定差异表达蛋白。使用基因本体论(GO)和京都基因与基因组百科全书(KEGG)预测差异表达蛋白的潜在功能。使用免疫组织化学(IHC)和蛋白质印迹(WB)验证差异表达蛋白。构建蛋白质-蛋白质相互作用(PPI)网络以描绘差异表达蛋白的调控网络。在正常和生精障碍的男性中鉴定出 3945 种蛋白质。其中,生精障碍男性中有 116 种蛋白差异表达:39 种上调,77 种下调。此外,我们发现这些差异表达蛋白主要涉及细胞成分,可能主要与剪接体、核糖体和甲状腺激素合成信号通路有关。剪接体和核糖体相关蛋白 YBX1、FBL 和 HNRNPU 下调。生精障碍男性睾丸组织的蛋白质组图谱与正常精子发生的男性不同。因此,YBX1、FBL 和 HNRNPU 等差异表达蛋白可能参与了精子发生功能障碍的病理过程。iTRAQ:相对和绝对定量同位素标记;GO:基因本体论;KEGG:京都基因与基因组百科全书;IHC:免疫组织化学;WB:蛋白质印迹;PPI:蛋白质-蛋白质相互作用;ICSI:胞浆内单精子注射;BP:生物学过程;CC:细胞成分;MF:分子功能;snoRNA:小核仁 RNA;snRNA:小核 RNA;LC-MS/MS:液相色谱和 MS/MS 分析;BSA:牛血清白蛋白;SD:精子发生功能障碍;micro-TESE:睾丸微观精子提取。