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Published (Version of Record)CC BY V4.0, Open Access
Journal article
1 H NMR spectroscopic characterisation of HepG2 cells as a model metabolic system for toxicology studies
Toxicology in vitro, Vol.99, 105881
2024
PMID: 38906200
Appears in Open Access via Read & Publish Agreements
Abstract
The immortalised human hepatocellular HepG2 cell line is commonly used for toxicology studies as an alternative to animal testing due to its characteristic liver-distinctive functions. However, little is known about the baseline metabolic changes within these cells upon toxin exposure. We have applied 1H Nuclear Magnetic Resonance (NMR) spectroscopy to characterise the biochemical composition of HepG2 cells at baseline and post-exposure to hydrogen peroxide (H2O2). Metabolic profiles of live cells, cell extracts, and their spent media supernatants were obtained using 1H high-resolution magic angle spinning (HR-MAS) NMR and 1H NMR spectroscopic techniques. Orthogonal partial least squares discriminant analysis (O-PLS-DA) was used to characterise the metabolites that differed between the baseline and H2O2 treated groups. The results showed that H2O2 caused alterations to 10 metabolites, including acetate, glutamate, lipids, phosphocholine, and creatine in the live cells; 25 metabolites, including acetate, alanine, adenosine diphosphate (ADP), aspartate, citrate, creatine, glucose, glutamine, glutathione, and lactate in the cell extracts, and 22 metabolites, including acetate, alanine, formate, glucose, pyruvate, phenylalanine, threonine, tryptophan, tyrosine, and valine in the cell supernatants. At least 10 biochemical pathways associated with these metabolites were disrupted upon toxin exposure, including those involved in energy, lipid, and amino acid metabolism. Our findings illustrate the ability of NMR-based metabolic profiling of immortalised human cells to detect metabolic effects on central metabolism due to toxin exposure. The established data sets will enable more subtle biochemical changes in the HepG2 model cell system to be identified in future toxicity testing.
Details
- Title
- 1 H NMR spectroscopic characterisation of HepG2 cells as a model metabolic system for toxicology studies
- Authors/Creators
- Maren JinksEmily C DaviesBerin A BoughtonSamantha LodgeGarth L Maker
- Publication Details
- Toxicology in vitro, Vol.99, 105881
- Publisher
- Elsevier Ltd.
- Number of pages
- 10
- Grant note
This paper is financially supported by an Australian Government Research Training Program (RTP) Scholarship to Maren Jinks. We also gratefully acknowledge the technical support for nuclear magnetic resonance analyses from Dr. Philipp Nitschke and Dr. Drew Hall at the Australian National Phenome Centre, Murdoch, Western Australia. The graphical abstract and Fig. 4 were created with BioRender.com
- Identifiers
- 991005707062507891
- Copyright
- © 2024 The Author(s).
- Murdoch Affiliation
- School of Medical, Molecular and Forensic Sciences; Centre for Computational and Systems Medicine
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- Citation topics
- 2 Chemistry
- 2.211 Mass Spectrometry
- 2.211.990 Metabolomics
- Web Of Science research areas
- Toxicology
- ESI research areas
- Pharmacology & Toxicology