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1H NMR spectroscopic characterisation of HepG2 cells as a model metabolic system for toxicology studies.
Jinks, Maren; Davies, Emily C; Boughton, Berin A; Lodge, Samantha; Maker, Garth L.
Afiliación
  • Jinks M; Australian National Phenome, Health Futures Institute, Harry Perkins Building, Murdoch University, Perth, WA 6150, Australia; Centre for Computational and Systems Medicine, Health Futures Institute, Harry Perkins Building, Murdoch University, Perth, WA 6150, Australia; Medical, Molecular and Forensi
  • Davies EC; Centre for Computational and Systems Medicine, Health Futures Institute, Harry Perkins Building, Murdoch University, Perth, WA 6150, Australia; Medical, Molecular and Forensic Sciences, Murdoch University, 90 South Street, Murdoch, WA 6150, Australia.
  • Boughton BA; Australian National Phenome, Health Futures Institute, Harry Perkins Building, Murdoch University, Perth, WA 6150, Australia; Centre for Computational and Systems Medicine, Health Futures Institute, Harry Perkins Building, Murdoch University, Perth, WA 6150, Australia; La Trobe Institute for Sustain
  • Lodge S; Australian National Phenome, Health Futures Institute, Harry Perkins Building, Murdoch University, Perth, WA 6150, Australia; Centre for Computational and Systems Medicine, Health Futures Institute, Harry Perkins Building, Murdoch University, Perth, WA 6150, Australia.
  • Maker GL; Centre for Computational and Systems Medicine, Health Futures Institute, Harry Perkins Building, Murdoch University, Perth, WA 6150, Australia; Medical, Molecular and Forensic Sciences, Murdoch University, 90 South Street, Murdoch, WA 6150, Australia. Electronic address: g.maker@murdoch.edu.au.
Toxicol In Vitro ; 99: 105881, 2024 Aug.
Article en En | MEDLINE | ID: mdl-38906200
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectroscopía de Protones por Resonancia Magnética / Peróxido de Hidrógeno Límite: Humans Idioma: En Revista: Toxicol In Vitro Asunto de la revista: TOXICOLOGIA Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectroscopía de Protones por Resonancia Magnética / Peróxido de Hidrógeno Límite: Humans Idioma: En Revista: Toxicol In Vitro Asunto de la revista: TOXICOLOGIA Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido