摘要:为探究邻苯二甲酸丁基苄酯(butylbenzylphthalate,BBP)对小鼠神经的毒性作用,进行了小鼠体外毒理学研究。首先用不同浓度的邻苯二甲酸丁基苄酯染毒神经模型细胞-N2a神经瘤细胞,通过噻唑蓝比色法(MTT),Hoechst33258染色实验评价邻苯二甲酸丁基苄酯的细胞毒效应;通过对染毒细胞氧自由基(ROS)、丙二醛(MDA)、还原型谷胱甘肽(GSH)含量的检测来探究BBP对小鼠神经瘤细胞的氧化损伤效应。随着BBP浓度的不断增高,细胞的MTT值逐渐变小,当BBP的浓度达到10g·L-1时,MTT实验结果与对照组出现显著性差异;Hoechst33258染色结果显示:高浓度的BBP导致细胞核呈现出不规则状态,出现了凋亡小体;随着BBP染毒浓度的升高N2a细胞中的ROS水平和MDA含量逐渐上升,分别在0.16g·L-1和10g·L-1开始与对照组相比出现了显著性的差异p〈0.05);而GSH系数呈现下降趋势,在0.32g·L。时开始出现显著性差异p〈0.05)。实验结果表明高浓度的邻苯二甲酸丁基苄酯可以导致神经瘤细胞的凋亡,并产生氧化损伤效应。
To study the toxicity of butyl benzyl phthalate (BBP) on nerve cells. Mouse neuroblastoma (N2a) cells were exposed in vitro to different concentrations of BBP. Then the toxicity of BBP on N2a cells were evaluated by MTT, Hoechst 33258 test. Furthermore the reactive oxygen species (ROS) level, malondialdehyde (MDA) and reduced glutathione (GSH) contents were measured for studying on the effects of oxidative dam- age induced by BBP. It was showed that the (OD) value of MTT assay decreased with the increasing of BBP concentrations and was significantly difference from the control at dose of 10 g·L-1. It was found that the nu- cleus irregular degree increased and appeared apoptotic bodies at high dose of BBP by Hoeehst 33258 staining. Along with the increasing of BBP concentrations, the enhancement of ROS level and MDA content at N2a cells were observed. There was significantly difference at dose of 0.16 g·L-1 and 10 g·L-1 respectively (p 〈 0.05). And GSH content decreased as exposure of BBP and was significantly difference from the control at dose of 0.32 g·L-1. These results suggested that high concentration of BBP could trigger off apoptosis and induce oxidative damage to N2a cells.