研究报告
江平.节球藻毒素诱导鲫鱼巨噬细胞凋亡的机理研究[J].环境科学学报,2014,34(5):1344-1350
节球藻毒素诱导鲫鱼巨噬细胞凋亡的机理研究
- Apoptosis induction of nodularin in Carassius auratus macrophages
- 摘要：选用杂食性鲤科鲫鱼（Carassius auratus）为材料，采用离体细胞培养诱导方法，研究了节球藻毒素在低剂量暴露条件下对鲫鱼巨噬细胞的毒效应.流式细胞仪结果证实，节球藻毒素可以诱导细胞出现凋亡峰，阻滞细胞G0/G1期和S期，扰乱细胞周期.1、10、100 μg·L-1节球藻毒素诱导12 h后，细胞凋亡率分别达到17.37%、27.59%、61.64%，而空白组的仅为3.72%.结果表明，100 μg·L-1的节球藻毒素处理12 h后，胞内钙离子浓度比空白组提高了76.9%，而线粒体膜电位则下降了35.8%，氧化应激产物活性氧（ROS）和丙二醛（MDA）含量则分别为空白组的1.86、2.94倍，Caspase-3和Caspase-9酶活性升高为空白组的2.89、3.73倍，而Caspase-8的酶活性仅比空白组的升高了33.3%，上述细胞凋亡线粒体通路指标均呈现明显的剂量-效应特征.综上所述，节球藻毒素可以诱导鲫鱼巨噬细胞发生凋亡，表现为线粒体依赖型，进而可能影响鱼类免疫系统.
- Abstract：The apoptotic responses induced by low dose nodularin on isolated macrophages from Carassius auratus were explored. Flow cytometric results showed that nodularin-treated macrophages had sub-G1 DNA peaks. Nodularin could block cells in G0/G1 and S phase and disturb cell cycle. The percentages of apoptotic cells incubated with 1, 10 and 100 μg·L-1 nodularin for 12 h reached 17.37%, 27.59% and 61.64%, respectively, while controls showed low rates of apoptosis (3.72%). The concentrations of intracellular Ca2+ for 100 μg·L-1 exposure group increased by 76.9% in comparison with the control group, while the mitochondrial transmembrane potential (MMP) decreased by 35.8%. The doses of reactive oxygen species (ROS) and malondialdehyde (MDA) for 100 μg·L-1 exposure group were 1.86 and 2.94 times that of the control group. After 100 μg·L-1 nodularin exposed for 12 h, the activity of Caspase-3 and Caspase-9 were 2.89 and 3.73 times that of the control group, respectively. The activity of Caspase-8 in 100μg·L-1 nodularin induced group increased by 33.3%, compared to the control group. All the biochemical apoptosis indicators analyzed in this study clearly showed dose-dependent manners. In summary, all the results suggested that nodularin could induce macrophages apoptosis via the mitochondrial apoptotic pathway and destroy the immune response of fish.
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