Study on the mechanism of NRK-52E cell injured by high glucose using LC-MS metabolomics

Objective To explore the injury mechanism of rat renal tubular epithelial cell (NRK-52E) induced by high glucose by analyzing the alteration of endogenous metabolites using liquid chromatography tandem with mass spectrometry (LC-MS) metabolomics. Methods NRK-52E cell that has been cultured for 12 h in normal condition were randomly divided into low glucose group (LG, 5.56 mmol/L, n=6) and high glucose group (HG, 25 mmol/L, n=6). After being cultured for 48 h, the metabolites from NRK-52E cell of LG and HG group were analyzed by LC-MS and the cell morphology was observed by optical microscope . The principal component analysis (PCA) and partial least squares to latent structure-discriminant analysis (PLS-DA) were performed using Mass Profile Professional (MPP) software to discriminant potential biomarkers. The structures of potential biomarkers were identified using METLIN data. Results The morphology of NRK-52E cells showed that the high- glucose environment was cytotoxic to NRK-52E cells. The results of metabolomics showed that the levels of leucine, hypoxanthine, ceramide (d18:0/12:0, d20:0/16:0, d18:0/16:0, t18:0/16:0, d18:0/14:0), p hosphatidylserine (16:0/16:0, 14:0/12:0, 20:0/0:0), phosphatidylinositol (17:2/20:2), lyso-phosphatidylethanolamine (0:0/20:5, 0:0/20:2) and phosphatidylethanolamine (16:0/0:0) in NRK-52E cells of HG group were significantly increased when comparing with the LG group . Conclusion The injury of NRK-52E induced by high glucose are closely with the metabolism of amino acid, purine and phospholipid in the cells