脱氧雪腐镰刀菌烯醇对仔猪海马神经细胞凋亡及线粒体膜电位的影响

[目的]本试验旨在研究脱氧雪腐镰刀菌烯醇（deoxynivalenol，DON）对仔猪海马神经细胞的凋亡及其信号通路的影响。[方法]体外培养仔猪海马神经细胞至对数生长期，以不同质量浓度的DON（0、125、250、500、1 000和2 000 ng？mL-1）进行处理，采用倒置显微镜和透射电镜观察细胞的形态学变化及线粒体结构，CCK-8试剂盒检测细胞的存活率，流式细胞仪检测线粒体膜电位的变化，ELISA试剂盒检测caspase-3酶活性，荧光定量PCR（RT-qPCR）检测Bcl-2和Bax mRNA的表达水平。[结果]DON可显著抑制仔猪海马神经细胞存活率并诱导细胞发生明显的形态学变化，且具有剂量和时间效应；DON可降低线粒体膜电位，增强caspase-3酶活力，并通过上调Bax mRNA和下调Bcl-2 mRNA的表达，增加Bax/Bcl-2值。[结论]DON可通过引起线粒体功能障碍，激活caspases-3并调节Bcl-2家族基因，从而诱导仔猪海马神经细胞凋亡。
[Objectives] The aim of this experiment was to study the effects of deoxynivalenol(DON)on apoptosis and its signaling pathway in piglet hippocampal nerve cells. [Methods] Hippocampal nerve cells were cultured to logarithmic phage, then were exposed to DON at different concentrations(0, 125, 250, 500, 1 000, 2 000 ng？mL-1). The cellular morphological changes and mitochondria changes were determined by inverted biological microscope and electron microscope. The cellular viability and mitochondrial membrane potential were measured by ELISA and flow cytometry. The key regulators of cell apoptosis, including caspase-3 activity, Bcl-2 and Bax mRNA expression, were determined by ELISA and fluorescence quantitative PCR(RT-qPCR), respectively. [Results] DON treatment could inhibit cell proliferation, and induce significant morphological changes with dose and time dependence. DON caused mitochondrial damage by decreasing the mitochondrial membrane potential. Furthermore, DON also induced apoptosis by promoting the activation of caspases-3 and Bax/Bcl-2 ratio, as well as down-regulated Bcl-2 mRNA level and up-regulated Bax mRNA level. [Conclusions] DON-induced apoptosis in hippocampal nerve cells of piglets was caused by mitochondrial dysfunction and successive activation of caspases-3, and the regulation of Bcl-2 family genes