Dept. Food Safety, German Federal Institute for Risk Assessment, Max-Dohrn-Str. 8-10, 10589, Berlin, Germany.
Dept. Pesticides Safety, German Federal Institute for Risk Assessment, Max-Dohrn-Str. 8-10, 10589, Berlin, Germany.
Arch Toxicol. 2019 Feb;93(2):369-383. doi: 10.1007/s00204-018-2360-3. Epub 2018 Nov 22.
The antihistaminic drug methapyrilene was withdrawn from the market in 1979 because of hepatocarcinogenicity in rats. Since then, the drug has been used as a model hepatotoxin especially for transcriptomic analyses using material from in vivo studies. Much less transcriptomics data are available from in vitro studies, and no studies have investigated proteomic effects of methapyrilene in vitro. Thus, the present study was aimed to characterize the proteomic response of primary rat hepatocytes to methapyrilene, to broaden our knowledge on the molecular mechanisms of methapyrilene toxicity, and to compare the results of collagen sandwich-cultured hepatocytes to in vivo data. In vitro methapyrilene concentrations (0.39 µM, 6.25 µM, and 100 µM) were chosen to cover an in vivo-relevant range. Based on published pharmacokinetic data they correspond to concentrations in portal vein blood for previously in vivo-tested doses of methapyrilene, up to a concentration showing slight cytotoxicity. Analysis of proteomic alterations by two-dimensional gel electrophoresis and mass-spectrometric protein identification demonstrated consistent and concentration-dependent effects of methapyrilene, in particular on mitochondrial proteins. Data suggest substantial deregulation of amino acid and ammonia metabolism and effects on mitochondrial energy supply pathways. The effects identified in vitro concur well with into previous in vivo observations. Several effects, for example, the influence of methapyrilene on S-adenosylmethionine metabolism, have not been described previously. The data suggest that already non-toxic concentrations of methapyrilene alter components of the intermediary metabolism, such as branched-chain amino acid metabolism, as well as urea and tricarboxylic cycle enzymes. In summary, data substantially add to our knowledge on molecular mechanisms of methapyrilene hepatotoxicity at the protein level.
抗组胺药甲吡戊烯因在大鼠中具有致癌性而于 1979 年从市场上撤出。此后，该药物一直被用作肝毒素模型，特别是用于体内研究的转录组分析。来自体外研究的转录组学数据要少得多，并且没有研究调查甲吡戊烯在体外的蛋白质组学效应。因此，本研究旨在表征原代大鼠肝细胞中甲吡戊烯的蛋白质组学反应，拓宽我们对甲吡戊烯毒性的分子机制的认识，并将胶原三明治培养的肝细胞的结果与体内数据进行比较。选择体外甲吡戊烯浓度（0.39µM、6.25µM 和 100µM）来涵盖体内相关范围。根据已发表的药代动力学数据，它们对应于以前在体内测试的甲吡戊烯剂量的门静脉血液中的浓度，直到出现轻微细胞毒性的浓度。通过二维凝胶电泳和质谱蛋白质鉴定分析蛋白质组学变化表明，甲吡戊烯具有一致的、浓度依赖性的作用，特别是对线粒体蛋白。数据表明氨基酸和氨代谢的大量失调以及对线粒体能量供应途径的影响。体外识别的效果与之前的体内观察结果非常吻合。一些效应，例如甲吡戊烯对 S-腺苷甲硫氨酸代谢的影响，以前没有描述过。这些数据表明，即使是甲吡戊烯的非毒性浓度也会改变中间代谢物的成分，如支链氨基酸代谢以及尿素和三羧酸循环酶。总之，数据在蛋白质水平上大大增加了我们对甲吡戊烯肝毒性的分子机制的认识。
