Abstract
Diabetic nephropathy is characterized by the chronic loss of kidney function due to high glucose renal levels. HK-2 proximal tubular cells are good candidates to study this disease. The aim of this work was to study an in vitro model of high glucose-induced metabolic alterations in HK-2 cells to contribute to the pathogenesis of this diabetic complication. An untargeted metabolomics strategy based on CE-MS was developed to find metabolites affected under high glucose conditions. Intracellular and extracellular fluids from HK-2 cells treated with 25 mM glucose (high glucose group), with 5.5 mM glucose (normal glucose group), and with 5.5 mM glucose and 19.5 mM mannitol (osmotic control group) were analyzed. The main changes induced by high glucose were found in the extracellular medium where increased levels of four amino acids were detected. Three of them (alanine, proline, and glutamic acid) were exported from HK-2 cells to the extracellular medium. Other affected metabolites include Amadori products and cysteine, which are more likely cause and consequence, respectively, of the oxidative stress induced by high glucose in HK-2 cells. The developed CE-MS platform provides valuable insight into high glucose-induced metabolic alterations in proximal tubular cells and allows identifying discriminative molecules of diabetic nephropathy.
Highlights
The International Diabetes Federation reported in 2017 that 451 million people have diabetes mellitus and estimated 5 million deaths worldwide as a result of its complications
The purpose of this work was to develop a non-targeted metabolomic platform based on capillary electrophoresis-mass spectrometry (CE-MS)
Since arginase hydrolyses l-arginine, the decreased content in l-arginine we have found in proximal tubular HK-2 cells exposed to high glucose might be pathologically relevant because it might be due to increased arginase-2 activity
Summary
The International Diabetes Federation reported in 2017 that 451 million people have diabetes mellitus and estimated 5 million deaths worldwide as a result of its complications. According to WHO, diabetes mellitus encompasses metabolic alterations of several etiologies characterized by chronic hyperglycemia and disorders in the metabolism of proteins, carbohydrates, and fats, due to the defects in insulin secretion [2]. To find new strategies for the prevention, early detection, or the study of diabetic nephropathy progress, it is necessary to know the discriminative molecules in order to identify the molecular mechanisms involved in this disease [4]. Metabolomics, especially in the non-targeted mode, offers great possibilities for studying the metabolome and the mechanisms implicated using different analytical techniques such as liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), or capillary electrophoresis-mass spectrometry (CE-MS)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
