Understanding of the nephrotoxicity induced by drug candidates is vital to drug discovery and development. Herein, an in situ metabolomics method based on air flow-assisted desorption electrospray ionization mass spectrometry imaging (AFADESI-MSI) was established for direct analysis of metabolites in renal tissue sections. This method was subsequently applied to investigate spatially resolved metabolic profile changes in rat kidney after the administration of aristolochic acid I, a known nephrotoxic drug, aimed to discover metabolites associated with nephrotoxicity. As a result, 38 metabolites related to the arginine–creatinine metabolic pathway, the urea cycle, the serine synthesis pathway, metabolism of lipids, choline, histamine, lysine, and adenosine triphosphate were significantly changed in the group treated with aristolochic acid I. These metabolites exhibited a unique distribution in rat kidney and a good spatial match with histopathological renal lesions. This study provides new insights into the mechanisms underlying aristolochic acids nephrotoxicity and demonstrates that AFADESI-MSI-based in situ metabolomics is a promising technique for investigation of the molecular mechanism of drug toxicity.

了解药物候选物引起的肾毒性对药物发现和开发至关重要。在这里，原位建立了基于气流辅助解吸电喷雾电离质谱成像（AFADESI-MSI）的代谢组学方法，可直接分析肾脏组织切片中的代谢物。该方法随后被用于研究马兜铃酸I（一种已知的肾毒性药物）的给药后在大鼠肾脏中空间分辨的代谢谱变化，其目的是发现与肾毒性相关的代谢产物。结果，在用马兜铃酸I处理的组中，与精氨酸-肌酐代谢途径，尿素循环，丝氨酸合成途径，脂质，胆碱，组胺，赖氨酸和三磷酸腺苷代谢有关的38种代谢物发生了显着变化。代谢物在大鼠肾脏中表现出独特的分布，并且与组织病理学肾脏病变具有良好的空间匹配性。原位代谢组学是一种研究药物毒性分子机制的有前途的技术。