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目的通过观察亚慢性铅暴露对小鼠睾丸组织Y染色体基因Ddx3y蛋白表达的影响,为探讨铅的雄性生殖毒作用机制提供新的靶标依据。方法:昆明种小鼠48只,按体重将小鼠随机分为对照组,0.5、1.0和1.5 g/L醋酸铅染毒组。通过自然饮用含不同浓度醋酸铅的方式使小鼠染铅,连续染毒60 d。取小鼠睾丸组织,用苏木素-伊红(HE)染色法进行组织病理学观察,用Western blotting和免疫组化方法检测睾丸组织Ddx3y蛋白的表达及组织分布。结果与对照组比较,染铅组睾丸曲细精管内细胞排列松散、紊乱、层次不清,精原细胞层和初级精母细胞层之间间隙变宽,且随着染铅浓度的增加精子细胞明显减少。Western blotting检测结果显示,染铅组的小鼠睾丸组织中Ddx3y蛋白表达显著低于对照组,且有剂量-反应关系。免疫组化结果显示,随着染铅剂量的增加,睾丸组织中抗Ddx3y蛋白免疫反应明显降低。结论亚慢性铅暴露可抑制小鼠精子的发生、发育和下调睾丸组织Y染色体基因Ddx3y蛋白的表达。
Objective To observe the effects of subchronic lead exposure on the expression of Y chromosome gene Ddx3y in mouse testes and to provide a new target basis for exploring the mechanisms of male reproductive toxicity of lead. Methods: Forty-eight Kunming mice were randomly divided into control group, 0.5, 1.0 and 1.5 g / L lead acetate exposure groups. The mice were exposed to lead by naturally drinking different concentrations of lead acetate for 60 days. The testicular tissue of mice was taken out, the hematoxylin-eosin (HE) staining was used to observe the histopathology, and the expression and distribution of Ddx3y in testis were detected by Western blotting and immunohistochemistry. Results Compared with the control group, the cells in testicular seminiferous tubules of lead-exposed group were loosely arranged, disorganized and unclear. The gap between spermatogonia and primary spermatocyte layer became wider. With the increase of lead concentration, spermatid obviously decrease. The result of Western blotting showed that the expression of Ddx3y protein in testis tissue of the lead-exposed group was significantly lower than that of the control group, and there was a dose-response relationship. Immunohistochemical results showed that with the lead dose increased, testicular tissue anti-Ddx3y protein immunoreactivity was significantly reduced. Conclusion Subchronic lead exposure can inhibit spermatogenesis in mice and develop and down-regulate the expression of Y chromosome gene Ddx3y in testis.