大鼠精索静脉曲张睾丸组织的蛋白质组学研究  

Proteomics study in rat testes of varicocele

作　　者：贺情情 刘小彭[1] 范年丰[2] 杨晓健[1] 张浩[1] 吴晓[1] 张炎[1] 

机构地区：[1]中山大学附属第三医院泌尿外,广州510630 [2]中山大学附属第三医院信息科,广州510630

出　　处：《中华腔镜泌尿外科杂志（电子版）》2014年第2期51-54,共4页Chinese Journal of Endourology(Electronic Edition)

基　　金：国家自然科学基金(30972996)

摘　　要：目的精索静脉曲张是男性不育的首要病因,本课题利用蛋白组学技术筛选精索静脉曲张大鼠睾丸中表达变化显著蛋白,探索精索静脉曲张引起不育的可能分子机制。方法雄性Wistar大鼠16只,随机分为对照组和精索静脉曲张实验组。实验组大鼠施行左肾静脉部分结扎术来诱导产生精索静脉曲张,对照组行假手术作为阴性对照。30 d后,收集全部大鼠的左侧睾丸。采用原位末端脱氧核苷酸转移酶标记(Tunel)法检测两组大鼠生精细胞凋亡情况,进而比较两组生精细胞的凋亡指数。利用双向凝胶电泳技术(2-DE)寻找两组大鼠睾丸中表达有显著性差异的蛋白质点,然后进行基质辅助激光解析离子化飞行时间质谱鉴定(MALDI-TOF MS)。结果双向凝胶电泳结果显示21个蛋白质点在两组间存在显著性差异。在精索静脉曲张大鼠睾丸中有8个蛋白质点表达上升、13个下降,这些蛋白功能涉及蛋白质合成、加工、降解,基因表达调控,信号转导,能量代谢,凋亡,自由基代谢等。Tunel证实实验组大鼠睾丸中生精细胞凋亡指数显著高于对照组,且本实验筛选出14-3-3ε、hnRNPF以及LZTFL1三种蛋白与凋亡相关。结论蛋白组学筛选出精索静脉曲张大鼠睾丸差异表达蛋白,并进一步证实精索静脉曲张促进睾丸生精细胞凋亡,其中相关凋亡蛋白对揭示精索静脉曲张引起男性不育的分子机制提供了可能。Objective Varicocele was the leading cause of male infertility. To explore the potential mechanism of infertility caused by varicocele, we used the proteomics technology to identify the significantly differential proteins in testes of rat with varicocele. Methods Sixteen Wistar rats were randomly divided into two groups, including varicocele group and sham operation group. The partial left renal vein ligation was applied to the rats in varicocele group, and sham operation group acted as the control. 30 days later, all the left testes were harvested for further study. Tunel method was applied to detect the apoptosis of germ cells, and the apoptosis index （AI） of the two groups was compared. Two-dimensional electrophoresis （2-DE） was used to screen the significantly altered proteins spots between the two groups. Following, the protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF MS）. Results AI was significantly different between the two groups and obviously higher in varicocele group. After screening the images of 2-DE, we found twenty-one protein spots were distinct caused by varicocele, and the expression of 8 protein spots were increased while the rest were decreased. These altered proteins were involved in protein synthesis, protein processing, protein degradation, regulation of gene expression, signal transduction, metabolism, apoptosis and free radical metabolism. Apoptosis proteins such as 14-3-3e, hnRNPF and LZTFL1 were screened in our investigation. Conclusions Proteomics technology was a powerful approach to screen the altered protein in testis with varicocele. Furthermore, varicocele was confirmed to cause higher apoptosis index of germ cells. Apoptosis proteins screened in our investigation may assist us to reveal the potential molecular mechanism of infertility.

关 键 词：蛋白质组学 精索静脉曲张 不育 凋亡 

分 类 号：R697.24[医药卫生—泌尿科学]