Univ Rennes, INSERM, INRAE, CHU Rennes, Institut NuMeCan (Nutrition, Metabolism and Cancer), PREVITOX Network, F-35033, Rennes, France; Rennes University Hospital, Forensic Toxicology Laboratory, F-35033, Rennes, France.
Univ Rennes, INSERM, INRAE, CHU Rennes, Institut NuMeCan (Nutrition, Metabolism and Cancer), PREVITOX Network, F-35033, Rennes, France.
Toxicol Lett. 2021 Aug 1;346:1-6. doi: 10.1016/j.toxlet.2021.04.006. Epub 2021 Apr 16.
Amanitin poisonings are among the most life-threatening mushroom poisonings, and are mainly caused by the genus Amanita. Hepatotoxicity is the hallmark of amanitins, powerful toxins contained in these mushrooms, and can require liver transplant. Among amatoxins, α-amanitin is the most studied. However, the hypothesis of a possible metabolism of amanitins is still controversial in this pathophysiology. Therefore, there is a need of clarification using cutting-edge tools allowing metabolism study. Molecular network has emerged as powerful tool allowing metabolism study through organization and representation of untargeted tandem mass spectrometry (MS/MS) data in a graphical form. The aim of this study is to investigate amanitin metabolism using molecular networking. In vivo (four positive amanitin urine samples) and in vitro (differentiated HepaRG cells supernatant incubated with α-amanitin 2 μM for 24 h) samples were extracted and analyzed by LC-HRMS/MS using a Q Exactive™ Orbitrap mass spectrometer. Using molecular networking on both in vitro and in vivo, we have demonstrated that α-amanitin does not undergo metabolism in human. Thus, we provide solid evidence that a possible production of amanitin metabolites cannot be involved in its toxicity pathways. These findings can help to settle the debate on amanitin metabolism and toxicity.
鹅膏蕈碱中毒是最具致命性的蘑菇中毒之一，主要由鹅膏菌属引起。鹅膏蕈碱是这些蘑菇中含有的强效毒素，其肝毒性是其特征，可能需要进行肝移植。在鹅膏蕈碱中，α-鹅膏蕈碱研究最多。然而，在这种病理生理学中，关于鹅膏蕈碱可能代谢的假设仍存在争议。因此，需要使用先进的工具来阐明这一问题，这些工具允许进行代谢研究。分子网络已成为一种强大的工具，可以通过将靶向串联质谱（MS/MS）数据以图形形式组织和表示，来进行代谢研究。本研究旨在使用分子网络研究鹅膏蕈碱的代谢。对来自体内（四个阳性鹅膏蕈碱尿液样本）和体外（分化的 HepaRG 细胞上清液在 2 μM α-鹅膏蕈碱孵育 24 小时）的样本进行提取，使用 Q Exactive ™ Orbitrap 质谱仪通过 LC-HRMS/MS 进行分析。通过对体内和体外样本进行分子网络分析，我们证明了α-鹅膏蕈碱在人体内不会发生代谢。因此，我们提供了确凿的证据，证明鹅膏蕈碱代谢物的可能产生不能参与其毒性途径。这些发现有助于解决关于鹅膏蕈碱代谢和毒性的争论。
