有氧运动对雄性大鼠生精功能的影响及其睾丸iTRAQ定量蛋白组学分析

发布时间：2019-04-29 11:18

【摘要】：目的:探讨有氧运动对雄性大鼠生精功能的影响,并通过蛋白组学筛选睾丸中有生精调控作用的差异表达蛋白,为进一步研究其分子机制奠定基础。方法:24只SD雄性大鼠随机分为对照组和运动组,对照组不进行运动训练。运动组分为无负荷运动组和3%体重负荷运动组,每天连续游泳60 min,无负荷运动组为无负重游泳,3%体重负荷运动组大鼠负荷3%体重重物游泳,每周6次,运动共9周。大鼠末次运动24 h后进行精子计数、生殖激素测定和睾丸组织学分析,并将3%体重负荷运动组和对照组睾丸组织进行同重同位素相对与绝对定量(iTRAQ)蛋白组学分析。结果:3%体重负荷运动组大鼠精子浓度[(3.54±0.52)×10~6/ml]明显高于对照组[(2.12±0.43)×10~6/ml],P0.01;血清LH[(38.96±1.34)IU/L]和T[(21.36±0.53)nmol/L]也显著高于对照组[LH:(35.99±2.90)IU/L,T:(19.99±0.25)nmol/L,P均0.01];GnRH和FSH两组无显著差异[GnRH:(641.82±42.78)ng/L vs(623.95±41.44)ng/L,FSH:(22.29±2.31)IU/L vs(20.49±2.44)IU/ml,P均0.05]。无负荷运动组精子浓度、生殖激素水平与对照组相比均无显著变化。组织学分析显示3%体重负荷运动组大鼠生精上皮内细胞层数较对照组明显增多,成熟精子数增加。蛋白组学分析筛查到差异蛋白47个,其中上调蛋白37个,下调蛋白10个。经过生物信息学分析筛选出与生殖功能密切相关的显著差异表达蛋白5个,其中Anx A1、GPX3、Rimbp3、Dpy19l2表达上调,CYP17表达下调。KEGG通路富集分析显示,差异蛋白主要参与了细胞外基质-受体相互作用通路、蛋白质消化与吸收通路、黏着斑信号通路等。结论:有氧运动可能主要通过改变精子发生与分化相关的蛋白表达来提高机体的生精功能。
[Abstract]:Aim: to investigate the effect of aerobic exercise on spermatogenic function of male rats, and to screen the differentially expressed proteins with spermatogenic regulation in testis by proteomics, so as to lay a foundation for further study of its molecular mechanism. Methods: 24 male SD rats were randomly divided into two groups: control group and exercise group. The exercise group was divided into two groups: no-load exercise group and 3% weight-loading exercise group. The rats in the 3% weight-load exercise group had no weight-bearing swimming in the 60 min, exercise-free exercise group, and the rats in the 3% weight-load exercise group had a load of 3% weight swimming, six times a week, for a total of 9 weeks. The sperm count, reproductive hormone and testicular histology were measured 24 hours after the last exercise, and the testis tissues of the 3% exercise group and the control group were analyzed with the same weight isotope relative to the absolute quantitative (iTRAQ) proteomics. Results: the sperm concentration in the 3% body weight exercise group [(3.54 卤0.52) 脳 10~6/ml] was significantly higher than that in the control group [(2.12 卤0.43) 脳 10~6/ml], P 0.01; Serum LH [(38.96 卤1.34) IU/L] and T [(21.36 卤0.53) nmol/L] were also significantly higher than those in the control group [LH: (35.99 卤2.90) IU/L,T: (19.99 卤0.25) nmol/L,P]. There was no significant difference between GnRH and FSH groups [GnRH: (641.82 卤42.78) ng/L vs (623.95 卤41.44) ng/L,FSH:, (22.29 卤2.31) IU/L vs (20.49 卤2.44) IU/ml,P]. There were no significant changes in sperm concentration and reproductive hormone level between the non-load exercise group and the control group. Histological analysis showed that the number of intracytoplasmic layers of spermatogenic epithelium and the number of mature spermatozoa in the 3% body weight exercise group were significantly higher than those in the control group. 47 differential proteins were detected by proteomics analysis, including 37 up-regulated proteins and 10 down-regulated proteins. The expression of Anx A1, GPX3, Rimbp3, Dpy19l2 was up-regulated and the expression of CYP17 was down-regulated by bioinformatics analysis. KEGG pathway enrichment analysis showed that the expression of Dpy19l2 was up-regulated and the expression of Dpy19l2 was down-regulated. Differential proteins are mainly involved in extracellular matrix-receptor interaction pathway, protein digestion and absorption pathway, focal adhesion signal pathway and so on. Conclusion: aerobic exercise may improve spermatogenic function by changing the expression of spermatogenesis and differentiation-related proteins.
【作者单位】：北京体育大学运动康复系;安阳师范学院体育学院;黄淮学院生物与食品工程学院;福建农林大学生命科学学院;
【基金】：河南省科技计划(132300410084) 北京体育大学校自主课题(2016BS044)~~
【分类号】：R698.2

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