Metabolomics profiling of steatosis progression in HepaRG ® cells using sodium valproate
Metabolomics profiling of steatosis progression in HepaRG ® cells using sodium valproate
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DOI：
10.1016/j.toxlet.2017.12.015
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发表时间：
2018-04-01
影响因子：
3.5
通讯作者：
Covaci, Adrian
中科院分区：
文献类型：
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作者：
Cuykx, Matthias;Claes, Leen;Covaci, Adrian
非酒精性脂肪性肝病（NAFLD）是一种常见的药物性肝损伤（DILI）。虽然这个阶段的疾病是可逆的，但它可能导致由非酒精性脂肪性肝炎（NASH），纤维化和肝硬化引起的不可逆损伤。因此，NAFLD的评估是新药候选药物毒理学筛选的首要目标。在本研究中，采用液-液萃取法，随后采用液相色谱法和精确质谱法（LC-AM/MS），获得了参比毒物丙戊酸钠（NaVPA）在4种给药方案下在HepaRG（R）细胞中诱导的NAFLD代谢组学指纹图。严格的实验设计与稳健的检测方法相结合，应用于丙戊酸钠，验证了非靶向代谢组学在肝脏毒理学研究中的可能性，一致观察到暴露和对照细胞之间的独特模式，多变量分析选择了多达200个感兴趣的特征，揭示了标志性NAFLD生物标志物，如二酰甘油和甘油三酯积累和肉毒碱缺乏。初始毒性反应显示S-腺苷甲硫氨酸和单乙酰亚精胺水平升高，甘油三酯仅中度升高。已经观察到新的特异性毒性标志物，如亚精胺、肌酸和乙酰胆碱。所描述的实验设计为毒理学危害的机制研究提供了有价值的代谢组学平台，并确定了脂肪变性进展的新标志物。
Non-alcoholic Fatty Liver Disease (NAFLD) is a frequently encountered Drug-Induced Liver Injury (DILI). Although this stage of the disease is reversible, it can lead to irreversible damage provoked by non-alcoholic steatohepatitis (NASH), fibrosis and cirrhosis. Therefore, the assessment of NAFLD is a paramount objective in toxicological screenings of new drug candidates. In this study, a metabolomic fingerprint of NAFLD induced in HepaRG (R) cells at four dosing schemes by a reference toxicant, sodium valproate (NaVPA), was obtained using liquid-liquid extraction followed by liquid chromatography and accurate mass-mass spectrometry (LC-AM/MS). The combination of a strict design of experiment with a robust detection method, applied on sodium valproate, validated the possibilities of untargeted metabolomics in hepatic toxicological research.Distinctive patterns between exposed and control cells were consistently observed, multivariate analyses selected up to 200 features of interest, revealing hallmark NAFLD-biomarkers, such as diacylglycerol and triglyceride accumulation and carnitine deficiency. Initial toxic responses show increased levels of S-adenosylmethionine and mono-acetylspermidine in combination with only a moderate increase in triglycerides. New specific markers of toxicity have been observed, such as spermidines, creatine, and acetylcholine. The described design of experiment provides a valuable metabolomics platform for mechanistic research of toxicological hazards and identified new markers for steatotic progression.