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An untargeted metabolomic strategy based on liquid chromatography-mass spectrometry to study high glucose-induced changes in HK-2 cells
- Source :
- e_Buah Biblioteca Digital Universidad de Alcalá, instname
- Publication Year :
- 2019
- Publisher :
- Elsevier BV, 2019.
-
Abstract
- https://www.sciencedirect.com/science/article/pii/S002196731930247X?via%3Dihub<br />Diabetes mellitus is a major health concern nowadays. It is estimated that 40 % of diabetics are affected by diabetic nephropathy, one of the complications derived from high glucose blood levels which can lead to chronic loss of kidney function. It is now clear that the renal proximal tubule plays a critical role in the progression of diabetic nephropathy but research focused on studying the molecular mechanisms involved is still needed. The aim of this work was to develop a liquid chromatography-mass spectrometry platform to carry out, for the first time, the untargeted metabolomic analysis of high glucose-induced changes in cultured human proximal tubular HK-2 cells. In order to find the metabolites which were affected by high glucose and to expand the metabolite coverage, intra- and extracellular fluid from HK-2 cells exposed to high glucose (25 mM), normal glucose (5.5 mM) or osmotic control (5.5 mM glucose + 19.5 mM mannitol) were analyzed by two complementary chromatographic modes: hydrophilic interaction and reversed-phase liquid chromatography. Non-supervised principal components analysis showed a good distribution among the three groups of samples. Statistically significant variables were chosen for further metabolite identification. Different metabolic pathways were affected mainly those derived from amino acidic, polyol, and nitrogenous bases metabolism.
- Subjects :
- Cells
Metabolite
010402 general chemistry
01 natural sciences
Biochemistry
Cell Line
Analytical Chemistry
Diabetic nephropathy
chemistry.chemical_compound
Metabolomics
Tandem Mass Spectrometry
Liquid chromatography–mass spectrometry
Diabetes mellitus
Extracellular fluid
medicine
HK-2 cells
Humans
liquid chromatography-mass spectrometry
Chromatography, Reverse-Phase
Chromatography
diabetic nephropathy
010401 analytical chemistry
Organic Chemistry
Química
General Medicine
Metabolism
medicine.disease
metabolomics
0104 chemical sciences
Chemistry
multivariate analysis
Glucose
chemistry
Mannitol
Hydrophobic and Hydrophilic Interactions
Metabolic Networks and Pathways
Chromatography, Liquid
medicine.drug
Subjects
Details
- ISSN :
- 00219673
- Volume :
- 1596
- Database :
- OpenAIRE
- Journal :
- Journal of Chromatography A
- Accession number :
- edsair.doi.dedup.....e0d7f8d06558a184505b2a6eef890625
- Full Text :
- https://doi.org/10.1016/j.chroma.2019.03.009