以原代培养的大鼠睾丸支持细胞为研究对象,选取环境及生物体中均有检出的一种典型的羟基化PBDE——6-HO-BDE-137作为目标化合物,设置0、0.1、1、10μmol·L^-1 4个浓度梯度,应用噻唑蓝(MTT)试验、荧光显微镜观察和AnnexinV-FITC/PI双染流式细胞术,通过检测大鼠睾丸支持细胞增殖活性、细胞形态和细胞凋亡坏死率的变化,探讨其对支持细胞的损伤情况.研究发现,6-HO-BDE-137显著影响支持细胞的增殖活力,暴露24h,10μmol·L^-1浓度组细胞与对照组相比显著增殖(p〈0.05),随暴露时间的延长(至48h),10μmol·L^-1组转为增殖抑制;支持细胞形态也表现出不同程度的改变,10μmol·L^-1浓度组变化最为明显,有大量的细胞变圆飘起;随6-HO-BDE-137暴露浓度的升高,支持细胞死亡率增加,凋亡是支持细胞死亡的主要方式.研究结果提示:6-HO-BDE-137具有生殖毒性.
In order to evaluate the toxicities of 6-HO-BDE-137 to rat testicular Sertoli cells, four concentrations of 6-HO- BDE-137 (0, 0.1, 1, 10μmol·L^-1)had been chosen. The testicular Sertoli cells of SD rats were exposed in vitro for 24h and 48h. Cell death and proliferation were determined by flow cytometric analysis and 3-[4,5-dimethylthiazol-2yl]-2,5- diphenyltetrazolium bromide (MTT)assay, and fluorescent microscopy was used to examine the morphological changes following treatment with 6-HO-BDE-137. Results showed that 6-HO-BDE-137 could affect the cell viability of Sertoli cells. The cell viability was significantly induced when Sertoli cells were treated with 10μmol·L^-1 6-HO-BDE-137 after 24h exposure and was significantly inhibited after 48h exposure. Sertoli cells morphology also changed after 6-HO-BDE- 137 exposure. The morphologic change was most evident at the concentration of 10~mol "L-~ and lots of cells turned round and floated. The percentage of dead Sertoli cells also increased with the increase of exposure concentration. Apoptosis is the main form of the death of Sertoli cells. These results indicate that 6-HO-BDE-137 exposure can cause reproductive toxicity.