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征，用免疫组织化学方法结合光镜和电子显微镜观察了大鼠支持细胞胞浆动力蛋白和运动蛋白的定位。胞质动力蛋白定位于微管、外质特化、线粒体和内质网。而激动素的反应区域与胞浆动力蛋白相似。其反应强度呈阶段性变化。其次，用光镜和电子显微镜观察了内分泌干扰物双酚A对青春期前雄性大鼠支持细胞培养的影响。在细胞培养中分别加入10、100和1000pg/ml的双酚A。观察到变性支持细胞的数量随剂量和时间的变化。与对照组比较，差异有统计学意义(P&lt；0.05)。变性的支持细胞表现为大的空泡、电子致密结构等。支持细胞变性程度以1000pg/ml浓度组最大，作用6d后最明显。这一发现表明，双酚-A对培养中的Sertoli细胞有直接影响。第三，研究了双酚A对小鼠睾丸的影响。成年雄性大鼠灌胃双酚A 6天后，分别于8天和36天摘除睾丸。光镜下观察。结果显示，部分生精小管生精细胞发生受阻，提示双酚-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.