An untargeted metabolomic strategy based on liquid chromatography-mass spectrometry to study high glucose-induced changes in HK-2 cells

Bernardo-Bermejo, Samuel; Sánchez-López, Elena; Castro-Puyana, María; Benito, S.; Lucio-Cazaña, Javier; Marina, Marı́a Luisa

doi:10.1016/j.chroma.2019.03.009

Cited by 21 publications

(24 citation statements)

References 44 publications

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“…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. The other metabolite affected by high glucose in the intracellular fluid was pyroglutamic acid, which is a product of amino acid catabolism, which was also identified in our previous metabolomics study in HK-2 cells by LC-MS [7].…”

Section: Biological Interpretation Of the Resultssupporting

confidence: 63%

“…In metabolomics studies by CE-MS, correction to IS is common because of larger variability inherent to this technique [32,33]. Data were further normalized to the protein content as in previous contributions [7,34,35].…”

Section: Non-targeted Metabolomics Analysis Of An In Vitro Model Of Hmentioning

confidence: 99%

“…Thus, the human proximal tubular HK-2 cell line is a good candidate for studying how high glucose-induced changes contribute to the genesis and progression of diabetic nephropathy. In fact, our research group has previously performed the first untargeted metabolomics analysis of HK-2 cells, which enabled us to obtain relevant information about diabetic nephropathy using LC-MS [7]. In addition, So et al carried out a study about diabetes on HK-2 cells using a proteomic approach [8].…”

Section: Introductionmentioning

confidence: 99%

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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

et al. 2020

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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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Section: Biological Interpretation Of the Resultssupporting

confidence: 63%

Section: Non-targeted Metabolomics Analysis Of An In Vitro Model Of Hmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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

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“…edu/) was used for peak exaction, data baseline filtering, baseline calibration, peak alignment, deconvolution analysis, peak identification, and peak integration 47,48 . The metabolites were filtered by removing exogenous compounds, such as drugs or compounds of plant origin, on the basis of their likelihood to be present in biological samples 49 .…”

Section: Scientific Reports |mentioning

confidence: 99%

Integrated LC–MS and GC–MS-based untargeted metabolomics studies of the effect of azadirachtin on Bactrocera dorsalis larvae

You

Qin

Zhang

et al. 2020

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Azadirachtin exhibits excellent bioactivities against several hundred arthropods. However, current knowlege of its biochemical effect on B. dorsalis larvae is not deep enough. In this study, integrated LC-MS and GC-MS-based untargeted metabolomics were used to analyze the changes of endogenous metabolites and the biochemical effects of azadirachtin on B. dorsalis larvae. Azadirachtin has excellent bioactivities against B. dorsalis larvae in this study, leading to a longer developmental duration, lower survival rate, and low pupa weight. The effect of azadirachtin was investigated on a total of 22 and 13 differentially abundant metabolites in the LC-MS and GC-MS-based metabolomics results, are selected respectively. Pathway analysis indicated that 14 differentially enriched metabolic pathways, including seven influential pathways, are worthy of attention. Further integrated key metabolic pathway analysis showed that histidine metabolism, d-glutamine and d-glutamate metabolism, biotin metabolism, ascorbate and aldarate metabolism, pentose and glucuronate interconversions, and alanine, aspartate and glutamate metabolism in B. dorsalis larvae are significantly relevant pathways affected by azadirachtin. Although extrapolating the bioactivity results in this study to the practical project of B. dorsalis pest management in the field has limitations, it was found that azadirachtin has a significant effect on the primary metabolism of B. dorsalis larvae.Bactrocera dorsalis is a destructive polyphagous and invasive insect pest of tropical and subtropical fruits and vegetables; this oriental fruit fly has been found to attack many types of commercial fruits and a wide variety of agricultural products 1 . Azadirachtin exhibits excellent bioactivities against agricultural, forestry, medical, and veterinary arthropods 2-4 . However, studies on the effects of azadirachtin on B. dorsalis are scarce. Azadirachtin is the main active ingredient in neem. It was reported that neem derivatives are ineffective when used as toxic bait for tephritid fruit flies 5 . Several studies reported that neem seed kernel extracts and azadirachtin deters oviposition of B. dorsalis adults 6,7 . Neem leaf dust significantly reduced the longevity and fertility of B. dorsalis adults by blocking ovarian development 8 . Neem extract could effectively reduce the fecundity, fertility, and post-embryonic development of freshly emerged B. dorsalis flies 9 . However, we found no previous studies in the literature on the activity of azadirachtin against the larvae of B. dorsalis.The biological effects of azadirachtin include impacts on egg-laying behavior, feeding behavior, growth and metamorphosis, reproduction, activity, and histopathology 10 . The mode of action of azadirachtin against lepidopteran insects can be explained, in part, by effects on digestive enzymes, NADPH cytochrome reductase, and cholinesterase 11 . The physiological effects of azadirachtin include direct inhibition of cell division and protein synthesis 12 . The indirect effects of blocki...

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“…Therefore, water containing 0.2 % formic acid with 10 mM ammonium formate (solvent A) and acetonitrile containing 0.2 % formic acid with 2 mM ammonium formate (solvent B) were employed using the same elution gradient. These mobile phases also showed to give good metabolite coverage for food and biological samples in previous works by our research team [27,28]. Under these conditions, the ionization was successful, and 280 and 223 molecular features were detected in positive and negative modes, respectively.…”

Section: Metabolomics Analysis Of Coffee Samples By Hilic-msmentioning

confidence: 58%

Untargeted HILIC-MS-Based Metabolomics Approach to Evaluate Coffee Roasting Process: Contributing to an Integrated Metabolomics Multiplatform

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An untargeted metabolomics strategy using hydrophilic interaction chromatography-mass spectrometry (HILIC-MS) was developed in this work enabling the study of the coffee roasting process. Green coffee beans and coffee beans submitted to three different roasting degrees (light, medium, and strong) were analyzed. Chromatographic separation was carried out using water containing 10 mM ammonium formate with 0.2 % formic acid (mobile phase A) and acetonitrile containing 10 mM ammonium formate with 0.2 % formic acid (mobile phase B). A total of 93 molecular features were considered from which 31 were chosen as the most statistically significant using variable in the projection values. 13 metabolites were tentatively identified as potential biomarkers of the coffee roasting process using this metabolomic platform. Results obtained in this work were complementary to those achieved using orthogonal techniques such as reversed-phase liquid chromatography-mass spectrometry (RPLC-MS) and capillary electrophoresis-mass spectrometry (CE-MS) since only one metabolite was found to be common between HILIC-MS and RPLC-MS platforms (caffeoylshikimic acid isomer) and other between HILIC-MS and CE-MS platforms (choline). On the basis of these results, an untargeted metabolomics multiplatform is proposed in this work based on the integration of the three orthogonal techniques as a powerful tool to expand the coverage of the roasted coffee metabolome.

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An untargeted metabolomic strategy based on liquid chromatography-mass spectrometry to study high glucose-induced changes in HK-2 cells

Cited by 21 publications

References 44 publications

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

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

Integrated LC–MS and GC–MS-based untargeted metabolomics studies of the effect of azadirachtin on Bactrocera dorsalis larvae

Untargeted HILIC-MS-Based Metabolomics Approach to Evaluate Coffee Roasting Process: Contributing to an Integrated Metabolomics Multiplatform

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