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目的研究热应激引起大鼠附睾早期微环境的变化。方法建立大鼠附睾热应激动物模型,42℃水浴30min,随后提取应激组和正常组附睾头部蛋白质,利用双向凝胶电泳分离、质谱鉴定大鼠附睾受热后差异变化的蛋白质;进一步通过生物信息学、Western blot和免疫组化等方法进行验证。结果蛋白质组学研究二维电泳图谱能检测到92个蛋白,经过质谱分析得出其中有24个蛋白具有意义,其中有12个蛋白上调,12个蛋白下调。热应激组与正常组有显著的差异蛋白质,这些具有意义的差异蛋白质经过免疫组化和Westem blot分析法也得出一致性的结论。结论热应激能导致附睾早期微环境变化,尤其是对附睾蛋白质差异表达以及对精子在附睾中的成熟过程有着显著的影响。
Objective To study the changes of early epididymal microenvironment induced by heat stress in rats. Methods The rat model of epididymal heat stress was established and treated with water bath at 42 ℃ for 30 min. The protein of epididymal head was extracted from the stress group and normal group. The proteins were separated by two-dimensional gel electrophoresis and identified by mass spectrometry. Bioinformatics, Western blot and immunohistochemistry. Results Proteomics Two-dimensional electrophoretograms were able to detect 92 proteins, of which 24 were found to be of significance by mass spectrometry. Twelve were up-regulated and 12 were down-regulated. There were significant differences between the heat stress group and the normal group, and the differences between these proteins were also concordant by immunohistochemistry and Westem blot analysis. Conclusion Heat stress can lead to early changes of epididymal microenvironment, especially the differential expression of epididymal proteins and sperm maturation in the epididymis has a significant impact.