Bourdin-Pintueles Alexandra, Galineau Laurent, Nadal-Desbarats Lydie, Dupuy Camille, Bodard Sylvie, Busson Julie, Lefèvre Antoine, Emond Patrick, Mavel Sylvie
UMR 1253 iBrain, Université de Tours, Inserm, Tours 37000, France.
CHRU de Tours, Service de Médecine Nucléaire In Vitro, Tours 37000, France.
J Proteome Res. 2021 Aug 6;20(8):3853-3864. doi: 10.1021/acs.jproteome.1c00145. Epub 2021 Jul 20.
Studying the metabolome of specific gestational compartments is of growing interest in the context of fetus developmental disorders. However, the metabolomes of the placenta and amniotic fluid (AF) are poorly characterized. Therefore, we present the validation of a fingerprinting methodology. Using pregnant rats, we performed exhaustive and robust extractions of metabolites in the AF and lipids and more polar metabolites in the placenta. For the AF, we compared the extraction capabilities of methanol (MeOH), acetonitrile (ACN), and a mixture of both. For the placenta, we compared (i) the extraction capabilities of dichloromethane, methyl -butyl ether (MTBE), and butanol, along with (ii) the impact of lyophilization of the placental tissue. Analyses were performed on a C18 and hydrophilic interaction liquid chromatography combined with high-resolution mass spectrometry. The efficiency and the robustness of the extractions were compared based on the number of the features or metabolites (for untargeted or targeted approach, respectively), their mean total intensity, and their coefficient of variation (% CV). The extraction capabilities of MeOH and ACN on the AF metabolome were equivalent. Lyophilization also had no significant impact and usefulness on the placental tissue metabolome profiling. Considering the placental lipidome, MTBE extraction was more informative because it allowed extraction of a slightly higher number of lipids, in higher concentration. This proof-of-concept study assessing the metabolomics and lipidomics of the AF and the placenta revealed changes in both metabolisms, at two different stages of rat gestation, and allowed a detailed prenatal metabolic fingerprinting.
在胎儿发育障碍的背景下，研究特定妊娠隔室的代谢组越来越受到关注。然而，胎盘和羊水（AF）的代谢组特征尚不明确。因此，我们展示了一种指纹图谱方法的验证。我们使用怀孕大鼠，对羊水中的代谢物以及胎盘中的脂质和极性更强的代谢物进行了全面而可靠的提取。对于羊水，我们比较了甲醇（MeOH）、乙腈（ACN）以及二者混合物的提取能力。对于胎盘，我们比较了（i）二氯甲烷、甲基叔丁基醚（MTBE）和丁醇的提取能力，以及（ii）胎盘组织冻干的影响。分析采用C18和亲水相互作用液相色谱结合高分辨率质谱进行。根据特征或代谢物的数量（分别针对非靶向或靶向方法）、它们的平均总强度及其变异系数（%CV）比较提取的效率和稳健性。甲醇和乙腈对羊水代谢组的提取能力相当。冻干对胎盘组织代谢组分析也没有显著影响和作用。考虑胎盘脂质组，MTBE提取提供的信息更多，因为它能提取出数量略多、浓度更高的脂质。这项评估羊水和胎盘代谢组学和脂质组学的概念验证研究揭示了大鼠妊娠两个不同阶段两种代谢的变化，并实现了详细的产前代谢指纹图谱分析。
