吸烟小鼠睾丸蛋白组差异研究：PEBP1对吸烟后小鼠睾丸精子发生功能影响的可能机制

发布时间：2018-03-30 18:39

  本文选题：吸烟　切入点：精子发生　出处：《上海交通大学》2012年硕士论文

【摘要】：吸烟被认为是影响精子可育性的重要危险因素,目前尚不清楚吸烟对精子可育性的影响机制。蛋白质组学是研究人员认识细胞功能最直接的工具,特定蛋白分子的鉴定能够帮助我们认识吸烟对精子不育影响的分子机制。 本研究首先通过睾丸的HE染色和精子数量、精子运动等参数的统计研究,发现吸烟后小鼠睾丸中精子数目明显减少,生精小管之间的支持细胞数量较正常小鼠也明显减少;附睾中分离的精子数量在吸烟后小鼠中明显减少,精子的运动能力受吸烟的影响较大。这说明了吸烟可能影响精子发生的功能,从而影响精子质量。 为了进一步探究吸烟对睾丸精子发生等功能影响的分子机制,我们通过比较蛋白质组学技术,发现了吸烟小鼠睾丸和正常小鼠睾丸之间差异表达的蛋白。我们用双向凝胶电泳(2 - DE)和质谱(MALDI - TOF - MS)技术鉴定吸烟小鼠睾丸和正常小鼠睾丸中差异表达的蛋白,发现了35种在吸烟小鼠睾丸中差异表达的蛋白,其中7种蛋白表达上调,其他的28种蛋白表达下调。 为了分析鉴定这35种蛋白,我们选择了DAVID服务器对他们聚类分析。这35种蛋白主要参与代谢,信号转导,细胞骨架组织和细胞运动维持等生物学过程。其中,在吸烟小鼠睾丸中表达下调的PEBP1可能与精子发生相关。我们分别用Western-Blot和qRT  PCR对PEBP1做了进一步分析。PEBP1蛋白和mRNA表达量在吸烟小鼠睾丸中均有所下降,这与前面的双向电泳结果一致。PEBP1是丝裂原激活的蛋白激酶(MAPK )信号转导通路的生理性抑制蛋白,吸烟小鼠睾丸中PEBP1表达的下降,导致MAPK信号途径的活跃。PEBP1可能成为临床上评估吸烟对生殖不良影响的分子标记,并提供吸烟对精子不育性的影响机制。
[Abstract]:Smoking is thought to be an important risk factor for sperm fertility, and it is not clear how smoking affects sperm fertility. Proteomics is the most direct tool for researchers to understand cell function. The identification of specific protein molecules can help us to understand the molecular mechanism of the effect of smoking on sperm sterility. In this study, the number of spermatozoa and Sertoli cells between seminiferous tubules were significantly decreased after smoking by means of HE staining, sperm number and sperm motility in testis of mice after smoking, and the number of Sertoli cells between seminiferous tubules was also significantly decreased compared with that of normal mice. The number of spermatozoa isolated from epididymis decreased significantly after smoking and the motility of spermatozoa was influenced by smoking, which indicated that smoking might affect the function of spermatogenesis and thus the quality of sperm. To further explore the molecular mechanism of the effects of smoking on testicular spermatogenesis, we compared proteomics techniques. The differentially expressed proteins were found between the testis of smoking mice and normal mice. The differential expression proteins were identified by two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MALDI-MS) in the testis of smoking mice and normal mice. 35 differentially expressed proteins were found in the testis of smoking mice, 7 of which were up-regulated and the other 28 were down-regulated. In order to analyze and identify the 35 proteins, we chose the DAVID server to cluster them. These 35 proteins are mainly involved in the biological processes of metabolism, signal transduction, cytoskeleton tissue and cell movement maintenance. The down-regulated expression of PEBP1 in the testis of smoking mice may be related to spermatogenesis. We further analyzed the expression of PEBP1 protein and mRNA in the testis of smoking mice by Western-Blot and qRT PCR. This is consistent with the previous results of two-dimensional electrophoresis. PEBP1 is a physiological inhibitor of mitogen-activated protein kinase (MAPK) signal transduction pathway, and the decrease of PEBP1 expression in the testis of smoking mice. The activation of MAPK signaling pathway. PEBP1 may be a molecular marker to evaluate the adverse effects of smoking on reproduction and provide a mechanism for the effect of smoking on sperm sterility.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

【引证文献】

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1 黄德伦;黄愉淋;付强;张明;;蛋白质组学在哺乳动物睾丸和精子发生中的应用[J];基因组学与应用生物学;2013年02期

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本文编号：1687138