Risk evaluation on endocrine disruptors using some testicular tissue culture systems

首先，为了了解支持细胞的形态和功能特点，细胞质动力蛋白和驱动蛋白在大鼠支持细胞的定位进行了研究，使用免疫组织化学与光镜和电镜。细胞质动力蛋白定位于微管，外质特化，线粒体和内质网。而驱动蛋白的反应区域与细胞质动力蛋白相似。其反应强度以阶段依赖性方式变化。其次，用光镜和电镜观察了内分泌干扰物双酚A对青春期前雄性大鼠睾丸支持细胞的影响。将双酚A分别以10、100和1000 pg/ml的浓度加入细胞培养物中。观察到支持细胞变性数量的剂量和时间依赖性变化。与对照组比较有显著性差异（p<0.05）。睾丸支持细胞在1000 pg/ml浓度时变性最严重，在处理6 d时变性最严重。这一发现表明，双酚-A显示出对培养中的支持细胞的直接影响。第三，进行了双酚A对睾丸影响的体内研究。成年雄性大鼠经口给予双酚A 6天后，在第8天和第36天切除睾丸。光镜观察。结果，在一些曲细精管中精子发生被破坏，表明双酚A对体内睾丸的影响。

First, in order to understand the morphological and functional characteristics of Sertoli cells, the localization of cytoplasmic dynein and kinesin in rat Sertoli cells was examined using immunohistochemistry with light and electron microscopies. Cytoplasmic dynein was localized in the microtubules, ectoplasmic specialization, mitochondria, and endoplasmic reticula. While, kinesin reacted in the regions similar to those of cytoplasmic dynein. Its reaction changed in intensity in a stage-dependent manner. Secondly, effects of bisphenol-A, one of endocrine disruptors, on the Sertoli cell culture from prepubertal male rats were examined by light and electron microscopies. Bisphenol-A was added to the cell culture at the concentration of 10, 100, and 1000 pg/ml, respectively. The dose-and time-dependent changes in the number of degenerated Sertoli cells were observed. They were significantly different from those in the control group (p<0.05). The degenerated Sertoli cells revealed some large vacuoles, electron-dense structures and so on. The maximum degenerative changes of Sertoli cells was observed at the concentration of 1000 pg/ml and at 6 days after treatment. This finding suggests that bisphenol-A shows a direct effect on Sertoli cells in culture. Thirdly, the in vivo study on the effects of bisphenol-A on testes was carried out. After the oral administration of bisphenol-A to adult male rats for 6 days, the testes were excised at 8 days and at 36 days. They were observed by light microscopy. As a result, spermatogenesis was disrupted in some seminiferous tubules, suggesting the influence of bisphenol-A on testes in vivo.