Université de Technologie de Compiègne, Centre de Recherche de Royallieu, Compiègne Cedex, France.
Toxicol Sci. 2013 Mar;132(1):8-20. doi: 10.1093/toxsci/kfs230. Epub 2012 Jul 27.
We used the recently introduced "metabolomics-on-a-chip" approach to test secondary drug toxicity in bioartificial organs. Bioartificial organs cultivated in microfluidic culture conditions provide a beneficial environment, in which the cellular cytoprotective mechanisms are enhanced, compared with Petri dish culture conditions. We investigated the metabolic response of HepG2/C3a cells exposed to flutamide, an anticancer prodrug, and hydroxyflutamide (HF), its active metabolite, in a microfluidic biochip. The cellular response was analyzed by (1)H nuclear magnetic resonance spectroscopy to identify cell-specific molecule-response markers. The metabolic response to flutamide results in a disruption of glucose homeostasis and in mitochondrial dysfunctions. This flutamide-specific metabolic response was illustrated by a reduction of the extracellular glucose and fructose consumptions and a general reduction of the tricarboxylic acid cycle activity leading to the reduction of the consumption of several amino acids. We also found a higher production of 3-hydroxybutyrate and lactate, and the reduction of the albumin production compared with controls. The toxic metabolic signature associated with the active metabolite HF was illustrated by a high-energy demand and an increase in several amino acid metabolism. Finally, for both molecules, the hepatotoxicity was correlated to the glutathione (GSH) metabolism illustrated by the levels of the 2-hydroxybutyrate and pyroglutamate productions and the increase of the glutamate and glycine productions. Thus, the entire set of results contributed to extract specific mechanistic toxic signatures and their relation to hepatotoxicity, which appeared consistent with literature reports. As new finding of HepG2/C3a cells hepatotoxicity, we propose a metabolic network with a related list of metabolite variations to describe the GSH depletion when followed by a cell death for the HepG2/C3a cells cultivated in our polydimethylsiloxane microfluidic biochips. Our findings illustrate the potential of metabolomics-on-a-chip as an in vitro alternative method for predictive toxicology.
我们使用最近引入的“芯片上代谢组学”方法来测试生物人工器官中的二次药物毒性。与 Petri 盘培养条件相比，在微流控培养条件下培养的生物人工器官提供了有益的环境，其中细胞细胞保护机制得到增强。我们研究了暴露于氟他胺（一种抗癌前药）和其活性代谢物羟基氟他胺（HF）的 HepG2/C3a 细胞在微流控生物芯片中的代谢反应。通过（1）H 核磁共振波谱分析鉴定细胞特异性分子反应标志物来分析细胞反应。氟他胺的代谢反应导致葡萄糖稳态紊乱和线粒体功能障碍。这种氟他胺特异性代谢反应表现为细胞外葡萄糖和果糖消耗减少，三羧酸循环活性普遍降低，导致几种氨基酸消耗减少。我们还发现 3-羟基丁酸和乳酸的产量增加，白蛋白的产量与对照相比降低。与对照相比，与活性代谢物 HF 相关的毒性代谢特征表现为高能量需求和几种氨基酸代谢增加。最后，对于这两种分子，肝毒性与谷胱甘肽（GSH）代谢相关，表现为 2-羟基丁酸和焦谷氨酸产量增加以及谷氨酸和甘氨酸产量增加。因此，整套结果有助于提取特定的机制毒性特征及其与肝毒性的关系，这与文献报道一致。作为 HepG2/C3a 细胞肝毒性的新发现，我们提出了一个代谢网络，其中包括代谢物变化的相关列表，以描述当 HepG2/C3a 细胞在我们的聚二甲基硅氧烷微流控生物芯片中培养时 GSH 耗竭后随之发生的细胞死亡。我们的研究结果表明，代谢组学在芯片上作为一种替代预测毒理学的体外方法具有潜力。
