A Non-Targeted Capillary Electrophoresis-Mass Spectrometry Strategy to Study Metabolic Differences in an In Vitro Model of High-Glucose Induced Changes in Human Proximal Tubular HK-2 Cells
Autores
Bernardo Bermejo, Samuel; Sánchez López, Elena; Castro Puyana, María; Benito Martínez, Selma; Lucio Cazaña, Francisco Javier de; [et al.]Identificadores
Enlace permanente (URI): http://hdl.handle.net/10017/41090DOI: doi:10.3390/molecules25030512
ISSN: 1420-3049
Fecha de publicación
2020Filiación
Universidad de Alcalá. Departamento de Cirugía, Ciencias Médicas y Sociales; Universidad de Alcalá. Departamento de Biología de Sistemas; Universidad de Alcalá. Departamento de Química Analítica, Química Física e Ingeniería QuímicaCita bibliográfica
Molecules, 2020, v. 25, n. 3, p. 512-
Palabras clave
diabetic nephropathy
human proximal tubular HK-2 cells
capillary electrophoresis-mass spectrometry
metabolomics
multivariate analysis
Tipo de documento
info:eu-repo/semantics/article
Versión
info:eu-repo/semantics/publishedVersion
Derechos
Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
Derechos de acceso
info:eu-repo/semantics/openAccess
Resumen
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.
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