Department of Urology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Asian J Androl. 2013 Jan;15(1):105-9. doi: 10.1038/aja.2012.64. Epub 2012 Aug 27.
Vasectomy is a simple and reliable method of male contraception. A growing number of men after vasectomy request vasectomy reversal due to various reasons. The pregnancy rate is lower than the patency rate after vasovasostomy and the pregnancy rate is time dependent. In this study, we evaluated the influence of reproductive tract obstruction on expression of epididymal proteins and their restoration after patency. Adult male Wistar rats were studied 30, 60 and 120 days after vasectomy, 30 days after vasovasostomy or after sham operations. Two-dimensional gel electrophoresis, mass-spectrometric technique, multidatabase search, Western blotting and real-time PCR were used to analyze the expression regulation of epididymal proteins. Total integrated intensity and total spot area of autoradiograms showed a consistent downward trend with time after obstruction, and this trend remained after patency. The intensity of the autoradiographic spots in three patency groups showed three trends: a downward trend, similar intensity and an upward trend compared with the correspondent obstruction group, respectively. Further verified experiments on human epididymis 2 (HE2), fertilization antigen-1 (FA-1), clusterin and PH20 demonstrated that compared with the correspondent obstruction group, the translation levels of HE2 and the mRNA transcription levels of HE2 showed an upward trend in patency groups, especially in the groups of obstruction for 60 days where the expression levels of HE2 were significantly upregulated after patency (P<0.05). Reproductive tract obstruction provokes a disregulation of gene expression in the epididymis and this disregulation remained after patency. Successful reversal may recover some proteins and the recovery is time dependent. Obstruction differentially alters mRNA transcription of different proteins and the content of proteins seemed to be easier to be influenced than the gene transcription.
输精管结扎术是一种简单可靠的男性避孕方法。由于各种原因，越来越多的输精管结扎术后男性要求进行输精管复通术。吻合术后的妊娠率低于通畅率，且妊娠率与时间有关。在这项研究中，我们评估了生殖道阻塞对附睾蛋白表达的影响及其再通后的恢复情况。研究雄性 Wistar 大鼠在输精管结扎后 30、60 和 120 天、吻合术后 30 天或假手术后，使用二维凝胶电泳、质谱技术、多数据库搜索、Western blot 和实时 PCR 分析附睾蛋白的表达调控。放射自显影的总积分强度和总斑点面积随阻塞后时间呈一致下降趋势，这种趋势在通畅后仍然存在。三个通畅组的放射自显影斑点强度显示出三种趋势：与相应的阻塞组相比，分别呈下降趋势、相似强度和上升趋势。进一步对人类附睾 2（HE2）、受精抗原 1（FA-1）、簇蛋白和 PH20 的验证实验表明，与相应的阻塞组相比，在通畅组中，HE2 的翻译水平和 HE2 的 mRNA 转录水平均呈上升趋势，尤其是在阻塞 60 天的组中，HE2 的表达水平在通畅后显著上调（P<0.05）。生殖道阻塞引起附睾基因表达失调，这种失调在通畅后仍然存在。成功的复通可能会恢复一些蛋白质，而且这种恢复是时间依赖的。阻塞对不同蛋白质的 mRNA 转录有不同的影响，蛋白质的含量似乎比基因转录更容易受到影响。
