Exploratory Metabolomic Analysis Based on Reversed-Phase Liquid Chromatography–Mass Spectrometry to Study an In Vitro Model of Hypoxia-Induced Metabolic Alterations in HK-2 Cells
Oxygen deficiency in cells, tissues, and organs can not only prevent the proper development of biological functions but it can also lead to several diseases and disorders. In this sense, the kidney deserves special attention since hypoxia can be considered an important factor in the pathophysiology...
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MDPI AG
2021-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/14/7399 |
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| author | Samuel Bernardo-Bermejo Elena Sánchez-López Lei Tan Selma Benito-Martínez Zhengjin Jiang María Castro-Puyana Francisco Javier Lucio-Cazaña María Luisa Marina |
| author_facet | Samuel Bernardo-Bermejo Elena Sánchez-López Lei Tan Selma Benito-Martínez Zhengjin Jiang María Castro-Puyana Francisco Javier Lucio-Cazaña María Luisa Marina |
| author_sort | Samuel Bernardo-Bermejo |
| collection | DOAJ |
| description | Oxygen deficiency in cells, tissues, and organs can not only prevent the proper development of biological functions but it can also lead to several diseases and disorders. In this sense, the kidney deserves special attention since hypoxia can be considered an important factor in the pathophysiology of both acute kidney injury and chronic kidney disease. To provide better knowledge to unveil the molecular mechanisms involved, new studies are necessary. In this sense, this work aims to study, for the first time, an in vitro model of hypoxia-induced metabolic alterations in human proximal tubular HK-2 cells because renal proximal tubules are particularly susceptible to hypoxia. Different groups of cells, cultivated under control and hypoxia conditions at 0.5, 5, 24, and 48 h, were investigated using untargeted metabolomic approaches based on reversed-phase liquid chromatography–mass spectrometry. Both intracellular and extracellular fluids were studied to obtain a large metabolite coverage. On the other hand, multivariate and univariate analyses were carried out to find the differences among the cell groups and to select the most relevant variables. The molecular features identified as affected metabolites were mainly amino acids and Amadori compounds. Insights about their biological relevance are also provided. |
| first_indexed | 2024-03-10T09:37:24Z |
| format | Article |
| id | doaj.art-826eab45dd1544b1b81e60605d3e5d8e |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T09:37:24Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-826eab45dd1544b1b81e60605d3e5d8e2023-11-22T03:57:54ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-012214739910.3390/ijms22147399Exploratory Metabolomic Analysis Based on Reversed-Phase Liquid Chromatography–Mass Spectrometry to Study an In Vitro Model of Hypoxia-Induced Metabolic Alterations in HK-2 CellsSamuel Bernardo-Bermejo0Elena Sánchez-López1Lei Tan2Selma Benito-Martínez3Zhengjin Jiang4María Castro-Puyana5Francisco Javier Lucio-Cazaña6María Luisa Marina7Departamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871 Madrid, SpainDepartment of Human Genetics, Leiden University Medical Center, 2333 ZA Leiden, The NetherlandsDepartamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871 Madrid, SpainDepartamento de Biología de Sistemas, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871 Madrid, SpainInstitute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou 510632, ChinaDepartamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871 Madrid, SpainDepartamento de Biología de Sistemas, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871 Madrid, SpainDepartamento de Química Analítica, Química Física e Ingeniería Química, Universidad de Alcalá, Ctra. Madrid-Barcelona Km. 33.600, 28871 Madrid, SpainOxygen deficiency in cells, tissues, and organs can not only prevent the proper development of biological functions but it can also lead to several diseases and disorders. In this sense, the kidney deserves special attention since hypoxia can be considered an important factor in the pathophysiology of both acute kidney injury and chronic kidney disease. To provide better knowledge to unveil the molecular mechanisms involved, new studies are necessary. In this sense, this work aims to study, for the first time, an in vitro model of hypoxia-induced metabolic alterations in human proximal tubular HK-2 cells because renal proximal tubules are particularly susceptible to hypoxia. Different groups of cells, cultivated under control and hypoxia conditions at 0.5, 5, 24, and 48 h, were investigated using untargeted metabolomic approaches based on reversed-phase liquid chromatography–mass spectrometry. Both intracellular and extracellular fluids were studied to obtain a large metabolite coverage. On the other hand, multivariate and univariate analyses were carried out to find the differences among the cell groups and to select the most relevant variables. The molecular features identified as affected metabolites were mainly amino acids and Amadori compounds. Insights about their biological relevance are also provided.https://www.mdpi.com/1422-0067/22/14/7399HK-2 cellshypoxialiquid chromatography–mass spectrometryuntargeted metabolomicsmultivariate analysis |
| spellingShingle | Samuel Bernardo-Bermejo Elena Sánchez-López Lei Tan Selma Benito-Martínez Zhengjin Jiang María Castro-Puyana Francisco Javier Lucio-Cazaña María Luisa Marina Exploratory Metabolomic Analysis Based on Reversed-Phase Liquid Chromatography–Mass Spectrometry to Study an In Vitro Model of Hypoxia-Induced Metabolic Alterations in HK-2 Cells International Journal of Molecular Sciences HK-2 cells hypoxia liquid chromatography–mass spectrometry untargeted metabolomics multivariate analysis |
| title | Exploratory Metabolomic Analysis Based on Reversed-Phase Liquid Chromatography–Mass Spectrometry to Study an In Vitro Model of Hypoxia-Induced Metabolic Alterations in HK-2 Cells |
| title_full | Exploratory Metabolomic Analysis Based on Reversed-Phase Liquid Chromatography–Mass Spectrometry to Study an In Vitro Model of Hypoxia-Induced Metabolic Alterations in HK-2 Cells |
| title_fullStr | Exploratory Metabolomic Analysis Based on Reversed-Phase Liquid Chromatography–Mass Spectrometry to Study an In Vitro Model of Hypoxia-Induced Metabolic Alterations in HK-2 Cells |
| title_full_unstemmed | Exploratory Metabolomic Analysis Based on Reversed-Phase Liquid Chromatography–Mass Spectrometry to Study an In Vitro Model of Hypoxia-Induced Metabolic Alterations in HK-2 Cells |
| title_short | Exploratory Metabolomic Analysis Based on Reversed-Phase Liquid Chromatography–Mass Spectrometry to Study an In Vitro Model of Hypoxia-Induced Metabolic Alterations in HK-2 Cells |
| title_sort | exploratory metabolomic analysis based on reversed phase liquid chromatography mass spectrometry to study an in vitro model of hypoxia induced metabolic alterations in hk 2 cells |
| topic | HK-2 cells hypoxia liquid chromatography–mass spectrometry untargeted metabolomics multivariate analysis |
| url | https://www.mdpi.com/1422-0067/22/14/7399 |
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