Metabolomic study of mouse embryonic fibroblast cells in response to autophagy based on high resolution gas chromatography-mass spectrometry

Autophagy is a lysosome-based degradation process, which is vital for cellular homeostasis. In recent years, its dysfunction has been reported to be closely related with various human diseases. Even though studies about autophagy from genomics and proteomics have been extensively conducted, the downstream metabolites and metabolic pathways associated with autophagy are rarely elucidated. In this study, we developed a high resolution gas chromatography-quadrupole orbitrap mass spectrometry (GCQ Orbitrap MS) method to conduct the metabolomic analysis of mouse embryonic fibroblast cells (MEFs) in response to autophagy by comparing wide type MEFs (WT MEFs) with Atg7-knockout MEFs (Atg7(-/-)MEFs) under starvation. After data acquisition and multivariate statistical analysis, 32 metabolites with significant difference (p < 0.05, VIP >1) were successfully identified by the library search and retention index (RI) calculation. Further metabolic pathway analysis proved that 34 metabolic pathways were significantly altered, which were mainly involved in carbohydrate, lipid, amino acid and other metabolism. Our results not only proved that carbohydrate metabolisms play an important role during autophagy along with amino acid metabolism and lipid metabolism, but also provided more autophagy-related target molecules, which contributes to the mechanism study of autophagy. (C) 2018 Elsevier B.V. All rights reserved.