Joré Céline, Loup Benoît, Garcia Patrice, Paris Anne-Christelle, Popot Marie-Agnès, Audran Michel, Bonnaire Yves, Varlet-Marie Emmanuelle, Bailly-Chouriberry Ludovic
Institut des Biomolécules Max Mousseron (IBMM), 15 Av. Charles Flahault, 34093 Montpellier, France; Laboratoire des Courses Hippiques (LCH), 15 rue de Paradis, 91370 Verrières-le-Buisson, France.
Laboratoire des Courses Hippiques (LCH), 15 rue de Paradis, 91370 Verrières-le-Buisson, France.
J Chromatogr A. 2017 Oct 27;1521:90-99. doi: 10.1016/j.chroma.2017.09.029. Epub 2017 Sep 14.
Erythropoiesis Stimulating Agents (ESAs) were developed for therapeutic purposes to stimulate red blood cell (RBC) production. Consequently, tissue oxygenation is enhanced as athlete's endurance and ESAs misuse now benefits doping. Our hypothesis is that most of ESAs should have similar mechanisms and thus have the same effects on metabolism. Studying the metabolome variations could allow suspecting the use of any ESAs with a single method by targeting their effects. In this objective, a metabolomic study was carried out on 3 thoroughbred horses with a single administration of 4.2μg/kg of Mircera, also called Continuous Erythropoiesis Receptor Activator (CERA). Blood and urine samples were collected from D to D and haematological parameters were followed throughout the study as plasmatic CERA concentration (ELISA). Urine and plasma metabolic fingerprints were recorded by Liquid Chromatography coupled to High Resolution Mass Spectrometry (LC-HRMS) in positive and negative mode. After preprocessing steps, normalized data were analyzed by multivariate statistics to build OPLS models. Hemoglobin concentration and hematocrit showed a significant increase after CERA administration unlike reticulocytes. CERA concentration showed a high intensity peak and then a slow decrease until becoming undetectable after D. Models built with multivariate statistics allow a discrimination between pre and post-administration plasma and urine samples until 74days after administration, i.e. 43days longer than ELISA method. By reducing and studying variables (ions), some potential candidate biomarkers were found.
促红细胞生成素（ESAs）是为治疗目的而开发的，用于刺激红细胞（RBC）生成。因此，随着运动员耐力的增强，组织氧合得到改善，而ESAs的滥用现在有利于兴奋剂使用。我们的假设是，大多数ESAs应该具有相似的机制，因此对新陈代谢有相同的影响。通过研究代谢组变化，有可能通过针对其作用，用单一方法怀疑任何ESAs的使用情况。为了实现这一目标，对3匹纯种马进行了一项代谢组学研究，单次给予4.2μg/kg的Mircera，也称为持续促红细胞生成素受体激活剂（CERA）。在给药后的不同时间点（D）采集血液和尿液样本，并在整个研究过程中跟踪血液学参数，同时检测血浆CERA浓度（酶联免疫吸附测定法）。通过液相色谱与高分辨率质谱联用（LC-HRMS）在正离子和负离子模式下记录尿液和血浆代谢指纹图谱。经过预处理步骤后，对归一化数据进行多变量统计分析以建立正交偏最小二乘法（OPLS）模型。与网织红细胞不同，给予CERA后血红蛋白浓度和血细胞比容显著增加。CERA浓度显示出一个高强度峰值，然后缓慢下降，直到给药后D天变得无法检测到。用多变量统计建立的模型能够区分给药前和给药后的血浆和尿液样本，直到给药后74天，即比酶联免疫吸附测定法长43天。通过减少和研究变量（离子），发现了一些潜在的候选生物标志物。
