Biochemical Toxicology, School of pharmaceutical sciences, Showa University, Tokyo, Japan.
J Toxicol Sci. 2011 Oct;36(5):601-11. doi: 10.2131/jts.36.601.
Ethylene glycol monomethyl ether (EGME) induces testicular lesion in rats and human. To investigate miRNAs expression in EGME testicular lesion, miRNA array assay and real-time RT-PCR analysis were conducted by using testis in rats treated with 50 and 2,000 mg/kg EGME for 6 and 24 hr. The expression of corresponding target gene for miRNAs was also examined. At 50 mg/kg, there were no changes in the gene expression and histopathological examination. At 2,000 mg/kg, slight decrease of phacytene spermatocytes with cell shrinkage and nucleus pyknosis at 6 hr and remarkable decrease (or cell death) of phacytene spermatocytes with Sertoli cell vacuolation at 24 hr were observed. After 24 hr, miR-449a and miR-92a decreased obviously and, miR-320, miR-134 and miR-188 increased, while only miR-760-5p increased after 6 hr. Above these miRNAs are reported to have an important role for spermatogenesis. The gene expression of Bcl-2, target for miR-449a, increased and therefore it is considered anti-apoptotic reaction has started in this stage. The expression of high mobility group AT-hook 2 (target of miR-92a) which regulates histone structure, was increased. Furthermore, histone deacethylase 4, targets for miR-320, was also affected. Above prohibiting apoptosis or activating epigenetic genes might be protective reaction to spermatocytes death under the miRNAs regulation in EGME testicular lesion.
乙二醇单甲醚(EGME)可诱导大鼠和人类睾丸损伤。为了研究 EGME 睾丸损伤中的 miRNA 表达,我们通过用 50 和 2000mg/kg EGME 处理大鼠 6 和 24 小时,进行了 miRNA 阵列分析和实时 RT-PCR 分析。还检查了 miRNA 的相应靶基因的表达。在 50mg/kg 时,基因表达和组织病理学检查没有变化。在 2000mg/kg 时,观察到 6 小时时精母细胞减数分裂减少,细胞收缩,细胞核固缩,24 小时时精母细胞减数分裂显著减少(或细胞死亡),并伴有支持细胞空泡化。24 小时后,miR-449a 和 miR-92a 明显减少,miR-320、miR-134 和 miR-188 增加,而只有 miR-760-5p 在 6 小时后增加。据报道,上述 miRNA 对精子发生具有重要作用。Bcl-2 的基因表达增加,其是 miR-449a 的靶基因,因此认为在该阶段已经开始了抗凋亡反应。高迁移率族 AT 钩 2(miR-92a 的靶基因)的表达增加,其调节组蛋白结构。此外,miR-320 的靶基因组蛋白去乙酰化酶 4 也受到影响。上述禁止细胞凋亡或激活表观遗传基因可能是在 EGME 睾丸损伤中 miRNA 调节下精母细胞死亡的保护反应。