Maneb alters central carbon metabolism and thiol redox status in a toxicant model of Parkinson's disease.
Maneb alters central carbon metabolism and thiol redox status in a toxicant model of Parkinson's disease.
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DOI：
10.1016/j.freeradbiomed.2020.11.028
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发表时间：
2021-01
影响因子：
7.4
通讯作者：
Roede JR
中科院分区：
文献类型：
--
作者：
Anderson CC;Marentette JO;Rauniyar AK;Prutton KM;Khatri M;Matheson C;Reisz JA;Reigan P;D'Alessandro A;Roede JR
二硫代氨基甲酸盐杀菌剂代森锰引起了人们的兴趣，由于农药的负面健康影响，以及其与帕金森病（PD）的关联越来越多的关注。我们的实验室以前曾报道过暴露于急性亚毒性水平MB的神经母细胞瘤细胞的不同表型变化，包括线粒体呼吸减少，乳酸动力学改变和代谢应激。在这项研究中，我们的目的是通过比较几种含巯基化合物及其对细胞能量代谢和巯基氧化还原节点的影响，进一步确定MB毒性的具体分子机制。采用细胞外通量分析和稳定同位素标记的示踪代谢组学来评估SK-N-AS人神经母细胞瘤细胞在急性暴露于一系列化合物后能量代谢的改变，所述化合物包括二硫代氨基甲酸盐（代森锰、代森钠、代森锌）和其他含巯基的小分子（谷胱甘肽、N-乙酰半胱氨酸）。这些研究表明，MB及其甲基化形式（MeDTC）是独特的毒物，对线粒体呼吸、增殖和糖酵解有显著影响。我们观察到MB通过氧化细胞谷胱甘肽和改变急性暴露后过氧化物氧还蛋白3（Prx 3，线粒体）的巯基氧化还原状态来显著影响细胞巯基氧化还原状态。氧化还原蛋白质印迹法揭示了MB的细胞Prx 3的特异性修饰，加强了MB可以优先靶向含有反应性半胱氨酸巯基的线粒体酶的论点。此外，稳定同位素示踪代谢组学证实了我们的能量评估，并表明，甲基溴暴露的结果在急性紊乱的中央碳代谢。具体来说，我们观察到细胞葡萄糖分流到戊糖磷酸途径和减少TCA中间体来自葡萄糖和谷氨酰胺。此外，我们报告了新的乳酸利用三氯乙酸富集和谷胱甘肽合成MB暴露后。总之，我们的结果进一步证实了MB通过巯基修饰发挥其毒性作用，并显著改变中心碳代谢。
The dithiocarbamate fungicide maneb (MB) has attracted interest due to increasing concern of the negative health effects of pesticides, as well as its association with Parkinson’s disease (PD). Our laboratory has previously reported distinct phenotypic changes of neuroblastoma cells exposed to acute, sub-toxic levels of MB, including decreased mitochondrial respiration, altered lactate dynamics, and metabolic stress. In this study, we aimed to further define the specific molecular mechanisms of MB toxicity through the comparison of several thiol-containing compounds and their effects on cellular energy metabolism and thiol redox nodes. Extracellular flux analyses and stable isotope labeled tracer metabolomics were employed to evaluate alterations in energy metabolism of SK-N-AS human neuroblastoma cells after acute exposure of an array of compounds, including dithiocarbamates (maneb, nabam, zineb) and other thiol-containing small molecules (glutathione, N-acetylcysteine). These studies revealed MB and its methylated form (MeDTC) as unique toxicants with significant alterations to mitochondrial respiration, proliferation, and glycolysis. We observed MB to significantly impact cellular thiol redox status by oxidizing cellular glutathione and altering the thiol redox status of peroxiredoxin 3 (Prx3, mitochondrial) after acute exposure. Redox Western blotting revealed a MB-specific modification of cellular Prx3, strengthening the argument that MB can preferentially target mitochondrial enzymes containing reactive cysteine thiols. Further, stable isotope tracer metabolomics confirmed our energetics assessments, and demonstrated that MB exposure results in acute derangement of central carbon metabolism. Specifically, we observed shunting of cellular glucose into the pentose-phosphate pathway and reduction of TCA intermediates derived from glucose and glutamine. Also, we report novel lactate utilization for TCA enrichment and glutathione synthesis after MB exposure. In summary, our results further confirm that MB exerts its toxic effects via thiol modification, and significantly transforms central carbon metabolism.
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影响因子：
3.3
作者：
Li, H;Dryhurst, G
通讯作者：
Dryhurst, G
影响因子：
13.6
作者：
Baron, S;Caplanusi, A;Smets, I
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