丙烯酰胺对原代培养大鼠大脑皮质神经元毒性作用的研究

Study on the toxic effect of acrylamide on primary cultured rat cerebral cortical neurons

目的:研究丙烯酰胺(acrylamide,ACR)对体外原代培养的大鼠大脑皮质神经元的毒性作用。方法:原代培养新生SD大鼠大脑皮质神经元,将培养至第7天的神经元分为空白对照组,丙烯酰胺不同浓度组。染毒后,四甲基偶氮唑盐比色法(MTT)检测各组皮质神经元的存活率,倒置显微镜观察各组皮质神经元的形态学变化,Hoechst33258染色法观察皮质神经元核形态的变化。结果:MTT结果显示各剂量ACR组的神经元存活率与对照组相比都显著降低,并且呈时间和剂量依赖性。ACR可造成皮质神经元突起萎缩,胞体肿胀,并可以诱导皮质神经元发生核固缩、碎裂,出现典型的凋亡特征,且随着浓度的升高,毒性作用加重。结论:ACR可对原代培养的大鼠大脑皮质神经元产生毒性损伤作用。它能抑制皮质神经元的存活,诱导神经元形态结构的改变、凋亡的发生。

Objective： To investigate the toxic effect of acrylamide on primary cultured rat cerebral cortex neurons.Methods： Cerebral cortex neurons were separated and cultured from new born Sprague-Dawley rats. When cultured at 7days,the cultured neurons were divided into control group and experimental groups which were treated with different concentrations of acrylamide. After treatment with acrylamide,the neuronal viability was measured by MTT assay,the morphological changes of neurons were observed under inverted microscope,and the morphological changes of nuclei were observed by Hoechst33258 assay. Results： The results of MTT assay showed that acrylamide significantly decreased the viability of cortex neurons in a time-dose-dependent manner compared with the control group. Acrylamide induced neurite atrophy and the swelling of the cell body. It also induced the karyopyknosis and karyorrhexis of cortex neurons,showing characteristics of apoptosis. The injury severity increased with the increasing of acrylamide concentration. Conclusion：Acrylamide could induce the toxic effects on the primary cultured rat cerebral cortex neurons. As a consequence,acrylamide inhibited the growth of cortex neurons and resulted in morphological changes of cortex neurons as well as apoptosis.