Acute Maneb Exposure Significantly Alters Both Glycolysis and Mitochondrial Function in Neuroblastoma Cells
Acute Maneb Exposure Significantly Alters Both Glycolysis and Mitochondrial Function in Neuroblastoma Cells
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DOI：
10.1093/toxsci/kfy116
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发表时间：
2018-09-01
影响因子：
3.8
通讯作者：
Roede, James R.
中科院分区：
文献类型：
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作者：
Anderson, Colin C.;Aivazidis, Stefanos;Roede, James R.
农药百草枯（PQ）和马涅布（MB）已被描述为帕金森病（PD）的环境危险因素，其机制与线粒体功能障碍和活性氧的产生有关。PQ和MB在小鼠模型和神经母细胞瘤细胞中的联合暴露已被用于进一步了解PD表型。MB作为一种氧化还原调节剂，通过烷基化蛋白质硫醇，并已被表征为抑制电子传递链的复合物III和解耦线粒体质子梯度。本研究的目的是利用海马细胞外通量平台分析人类神经母细胞瘤细胞中atp相关的呼吸和糖酵解。本研究采用急性亚毒性暴露的MB，揭示了MB介导的线粒体耗氧量在基线和最大呼吸时的降低，并抑制ATP合成和偶联效率。此外，mb处理的细胞显示出非线粒体呼吸和质子泄漏的增加。对线粒体燃料柔韧性的进一步研究显示，所有3种主要底物的燃料柔韧性都消失了，丙酮酸容量和谷氨酰胺依赖性都下降了。糖酵解功能分析显示，乳酸出口导致糖酵解酸化显著减少。在低至6 μ M的MB剂量滴定后，也观察到这种糖酵解参数的抑制作用，并且似乎依赖于二硫代氨基甲酸酯官能团，锰可能增强了这种作用。对细胞ATP和NAD水平的进一步研究显示，MB处理的细胞急剧下降。总之，MB显著影响好氧和厌氧能量的产生；因此，进一步表征MB对细胞能量学的影响可能有助于了解PD对多巴胺能神经元的特异性。
The pesticides paraquat (PQ) and maneb (MB) have been described as environmental risk factors for Parkinson's disease (PD), with mechanisms associated with mitochondrial dysfunction and reactive oxygen species generation. A combined exposure of PQ and MB in murine models and neuroblastoma cells has been utilized to further advance understanding of the PD phenotype. MB acts as a redox modulator through alkylation of protein thiols and has been previously characterized to inhibit complex III of the electron transport chain and uncouple the mitochondrial proton gradient. The purpose of this study was to analyze ATP-linked respiration and glycolysis in human neuroblastoma cells utilizing the Seahorse extracellular flux platform. Employing an acute, subtoxic exposure of MB, this investigation revealed a MB-mediated decrease in mitochondrial oxygen consumption at baseline and maximal respiration, with inhibition of ATP synthesis and coupling efficiency. Additionally, MB-treated cells showed an increase in nonmitochondrial respiration and proton leak. Further investigation into mitochondrial fuel flex revealed an elimination of fuel flexibility across all 3 major substrates, with a decrease in pyruvate capacity as well as glutamine dependency. Analyses of glycolytic function showed a substantial decrease in glycolytic acidification caused by lactic acid export. This inhibition of glycolytic parameters was also observed after titrating the MB dose as low as 6 mu M, and appears to be dependent on the dithiocarbamate functional group, with manganese possibly potentiating the effect. Further studies into cellular ATP and NAD levels revealed a drastic decrease in cells treated with MB. In summary, MB significantly impacted both aerobic and anaerobic energy production; therefore, further characterization of MB's effect on cellular energetics may provide insight into the specificity of PD to dopaminergic neurons.