Cargando…
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
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 con...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7037647/ https://www.ncbi.nlm.nih.gov/pubmed/31991659 http://dx.doi.org/10.3390/molecules25030512 |
_version_ | 1783500472733138944 |
---|---|
author | Bernardo-Bermejo, Samuel Sánchez-López, Elena Castro-Puyana, María Benito-Martínez, Selma Lucio-Cazaña, Francisco Javier Marina, María Luisa |
author_facet | Bernardo-Bermejo, Samuel Sánchez-López, Elena Castro-Puyana, María Benito-Martínez, Selma Lucio-Cazaña, Francisco Javier Marina, María Luisa |
author_sort | Bernardo-Bermejo, Samuel |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7037647 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-70376472020-03-11 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 Bernardo-Bermejo, Samuel Sánchez-López, Elena Castro-Puyana, María Benito-Martínez, Selma Lucio-Cazaña, Francisco Javier Marina, María Luisa Molecules Article 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. MDPI 2020-01-24 /pmc/articles/PMC7037647/ /pubmed/31991659 http://dx.doi.org/10.3390/molecules25030512 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Bernardo-Bermejo, Samuel Sánchez-López, Elena Castro-Puyana, María Benito-Martínez, Selma Lucio-Cazaña, Francisco Javier Marina, María Luisa 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 |
title | 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 |
title_full | 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 |
title_fullStr | 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 |
title_full_unstemmed | 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 |
title_short | 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 |
title_sort | 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 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7037647/ https://www.ncbi.nlm.nih.gov/pubmed/31991659 http://dx.doi.org/10.3390/molecules25030512 |
work_keys_str_mv | AT bernardobermejosamuel anontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT sanchezlopezelena anontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT castropuyanamaria anontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT benitomartinezselma anontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT luciocazanafranciscojavier anontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT marinamarialuisa anontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT bernardobermejosamuel nontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT sanchezlopezelena nontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT castropuyanamaria nontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT benitomartinezselma nontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT luciocazanafranciscojavier nontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells AT marinamarialuisa nontargetedcapillaryelectrophoresismassspectrometrystrategytostudymetabolicdifferencesinaninvitromodelofhighglucoseinducedchangesinhumanproximaltubularhk2cells |