Shiraishi Koji, Naito Katsusuke
Department of Urology, Yamaguchi University School of Medicine, Yamaguchi, Japan.
J Urol. 2007 Sep;178(3 Pt 1):1012-7; discussion 1017. doi: 10.1016/j.juro.2007.05.027. Epub 2007 Jul 16.
Oxidative stress is involved in male infertility. However, little is known about how it impairs spermatogenesis. We investigated the presence of oxidative stress in human testes by studying the generation of 4-hydroxy-2-nonenal modified proteins and expressions of proliferating cell nuclear antigen and p53.
A total of 47 testicular biopsies from patients with varicocele, obstructive azoospermia and idiopathic infertility were included. Localization and generation of 4-hydroxy-2-nonenal modified proteins were determined by immunohistochemistry and Western blotting. The expressions of proliferating cell nuclear antigen and p53 were assessed by Western blotting. The interaction between 4-hydroxy-2-nonenal modified proteins and p53 was examined by immunoprecipitation. Data were compared to clinicopathological parameters.
4-Hydroxy-2-nonenal modified proteins were strongly positive in spermatogonia, primary spermatocytes and Sertoli's cells, and generation was inversely correlated with expression of proliferating cell nuclear antigen. The expression of p53 was increased in testes with varicocele (p <0.01) and obstructive azoospermia (p <0.05), and there was a positive or inverse correlation with 4-hydroxy-2-nonenal modified proteins and proliferating cell nuclear antigen. Immunoprecipitated p53 was detected by anti-4-hydroxy-2-nonenal modified protein antibody.
4-Hydroxy-2-nonenal impairs the proliferation of germ cells through the up-regulation of p53 protein, especially in testes with varicocele. Modification by 4-hydroxy-2-nonenal might alter normal function and stabilization of p53 protein.
氧化应激与男性不育有关。然而，关于其如何损害精子发生却知之甚少。我们通过研究4-羟基-2-壬烯醛修饰蛋白的生成以及增殖细胞核抗原和p53的表达，来调查人类睾丸中氧化应激的存在情况。
纳入了47例精索静脉曲张、梗阻性无精子症和特发性不育患者的睾丸活检样本。通过免疫组织化学和蛋白质印迹法测定4-羟基-2-壬烯醛修饰蛋白的定位和生成情况。通过蛋白质印迹法评估增殖细胞核抗原和p53的表达。通过免疫沉淀法检测4-羟基-2-壬烯醛修饰蛋白与p53之间的相互作用。将数据与临床病理参数进行比较。
4-羟基-2-壬烯醛修饰蛋白在精原细胞、初级精母细胞和支持细胞中呈强阳性，其生成与增殖细胞核抗原的表达呈负相关。精索静脉曲张患者（p<0.01）和梗阻性无精子症患者（p<0.05）的睾丸中p53表达增加，且与4-羟基-2-壬烯醛修饰蛋白和增殖细胞核抗原呈正相关或负相关。用抗4-羟基-2-壬烯醛修饰蛋白抗体检测到免疫沉淀的p53。
4-羟基-2-壬烯醛通过上调p53蛋白损害生殖细胞的增殖，尤其是在精索静脉曲张患者的睾丸中。4-羟基-2-壬烯醛的修饰可能会改变p53蛋白的正常功能和稳定性。
