论文部分内容阅读
目的:利用“shotgun(鸟枪)”蛋白质组学策略鉴定精浆中弱精子症相关的蛋白质。方法:正常健康已生育和弱精子症男性各3例自愿捐献的精液标本,通过Percoll分离获取精浆,等量混合后在SDS-PAGE上进行分离,切胶取条带进行胶内酶解,抽提肽段后利用“shotgun”蛋白组学策略鉴定蛋白质。然后,将两组鉴定到的唯一肽段数≥2和唯一肽段数=1但肽段总数≥4的蛋白质对比分析。结果:正常生育男性和弱精子症男性精浆中共得到172种差异蛋白质,全部是有和无的关系。其中89种蛋白质存在于弱精子症患者,而正常生育者没有鉴定到;83种蛋白质存在于正常可生育者,而弱精子症患者没有鉴定到。这些差异蛋白质集中在信号转导和酶催化活性类。结论:通过分析差异蛋白质的功能,有10种蛋白质尤其重要,可能与弱精子症相关,这10种蛋白质为:膜联蛋白VI同工型2、白介素6受体β亚单位同工型1前体、相对分子质量为400000的蛋白质、胞质动力蛋白重链、α-辅肌动蛋白4、受体型酪氨酸蛋白磷酸酶η前体、维生素D结合蛋白前体、蛋白S100-A11、蛋白S100-A9和膜联蛋白A4。
OBJECTIVE: To identify weakly spermatozoal related proteins in seminal plasma using the “shotgun” proteomics strategy. Methods: Semen samples were collected from 3 normal volunteers who had normal fertility and weak spermatozoa. Seminal plasma was separated by Percoll, separated by SDS-PAGE in the same amount and separated by gel filtration. Proteins were identified using the “shotgun” proteomics strategy after extraction of peptides. Then, two groups of identified proteins with a unique number of peptides> = 2 and a unique peptide number = 1 but a total number of peptides> 4 were compared. Results: A total of 172 differentially expressed proteins were found in normal seminal plasma and asthenospermia male seminal plasma, all with and without relationship. Of these, 89 proteins were found in patients with asthenospermia, whereas normal births were not identified; 83 proteins were found in normal fertile subjects and in asthenospermia patients were not identified. These differential proteins are focused on signal transduction and enzymatic active classes. CONCLUSIONS: Ten proteins are of particular importance by analyzing the function of differential proteins and may be associated with asthenospermia. These ten proteins are annexin VI isoform 2, interleukin 6 receptor beta subunit 1 Body, a protein with a relative molecular mass of 400000, a cytoplasmic dynein heavy chain, α-actinin 4, a receptor tyrosine phosphatase η precursor, a vitamin D binding protein precursor, a protein S100-A11, Protein S100-A9 and Annexin A4.