少弱精子症患者精子候选差异蛋白Trx 2、TrxR 1的验证及在少精、弱精子症中的表达研究

目的:根据TMT技术筛选少弱精子症患者精子差异蛋白的结果,选取硫氧还蛋白2(thioredoxin 2,Trx 2)、硫氧还蛋白还原酶1(thioredoxin reductase 1,TrxR 1)进行验证,探讨二者在少精、弱精和少弱精子症中的表达变化及其意义。方法:收集105例少精子症组(O组)、150例弱精子症组(A组)、50例少弱精子症组(OA组)和106例正常精液男性(N组)精液,分离出精子,对少弱精子症进行串联质谱标签(Tandem Mass Tag,TMT)技术蛋白质组学分析,根据少弱精子症组的精子差异蛋白结果选取Trx 2、TrxR 1,通过免疫荧光和免疫印迹方法检测其在O组、A组、OA组的表达情况。结果:TMT技术蛋白质组学结果显示Trx 2为上调差异蛋白(为N组的1.31倍),TrxR 1为下调差异蛋白(为N组的0.82倍)。免疫荧光和免疫印迹结果显示O组、A组、OA组Trx 2表达显著高于N组(P0.05),O组、OA组TrxR 1的表达显著低于N组(P0.05)。二者在OA组的结果与蛋白质组学结果一致。结论:Trx 2、TrxR 1可能在少精、弱精及少弱精子症的发生中起着重要的作用,并有望成为少弱精子症患者精子的候选标志物及治疗靶点。

Verification of Candidate Differential Sperm Proteins Trx-2 and TrxR-1 in Oligoasthenospermia and Their Expression in the Oligospermia and Asthenospermia

ABSTRACT Objective: According to the results of TMT screening of sperm differential proteins in oligoasthenospermia patients, thioredoxin 2 (Trx 2) and thioredoxin reductase 1 (TrxR 1) were selected to verify their expression and significances in oligospermia, asthenospermia and oligoasthenospermia. Methods: Sperms were collected from 105 cases of oligospermia(O group), 150 cases of asthenospermia (A group), 50 cases of oligoasthenospermia group (OA group) and 106 cases of normal semen men group (N group) and separated for proteomic analysis by Tandem Mass Tag(TMT). Trx 2 and TrxR 1 were selected according to the results of differential proteins and detected their expression by immunofluorescence and Western blotting. Results: The proteomic results of sperm TMT showed that Trx 2 was up-regulated differential protein (1.31 times higher than that in group N), and TrxR 1 was down-regulated differential protein (0.82 times higher than that in group N). Immunofluorescence and Western blotting results showed that Trx 2 in group O, group A, group OA were significantly higher than that in the group N (P<0.05), while the expression of TrxR 1 in group O and group OA were significantly lower than that in the group N (P<0.05), which was consistent with the screening results. Conclusion: Trx 2 and TrxR 1 may play important roles in the pathogenesis of oligospermia, asthenospermia and oligoasthenospermia, and may become candidate markers and therapeutic targets for the oligoasthenospermia patients.