Hyperglycemia Stimulates the Irreversible Catabolism of Branched-Chain Amino Acids and Generation of Ketone Bodies by Cultured Human Astrocytes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Cell Culture
2.3. 1H-NMR Analysis
Cell Lysate Preparation
2.4. Protein Estimation
2.5. Cell Survival
2.6. Enzymatic Estimation of 3-Hydroxybutyrate Level
2.7. Statistical Analysis
3. Results
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Beard, E.; Lengacher, S.; Dias, S.; Magistretti, P.J.; Finsterwald, C. Astrocytes as Key Regulators of Brain Energy Metabolism: New Therapeutic Perspectives. Front. Physiol. 2022, 12, 825816. [Google Scholar] [CrossRef] [PubMed]
- Bélanger, M.; Allaman, I.; Magistretti, P.J. Brain Energy Metabolism: Focus on Astrocyte-Neuron Metabolic Cooperation. Cell Metab. 2011, 14, 724–738. [Google Scholar] [CrossRef] [PubMed]
- Bonvento, G.; Bolaños, J.P. Astrocyte-Neuron Metabolic Cooperation Shapes Brain Activity. Cell Metab. 2021, 33, 1546–1564. [Google Scholar] [CrossRef] [PubMed]
- Hamprecht, B.; Verleysdonk, S.; Wiesinger, H. Enzymes of Carbohydrate and Energy Metabolism. In Neuroglia; Kettenmann, H., Ransom, B.R., Eds.; Oxford University Press: Oxford, UK, 2004; ISBN 978-0-19-515222-7. [Google Scholar]
- Verkhratsky, A.; Semyanov, A. The Great Astroglial Metabolic Revolution: Mitochondria Fuel Astrocyte Homeostatic Support and Neuroprotection. Cell Calcium 2022, 104, 102583. [Google Scholar] [CrossRef] [PubMed]
- Daneman, R.; Prat, A. The Blood–Brain Barrier. Cold Spring Harb. Perspect. Biol. 2015, 7, a020412. [Google Scholar] [CrossRef]
- Pardridge, W.M. Brain Metabolism: A Perspective from the Blood-Brain Barrier. Physiol. Rev. 1983, 63, 1481–1535. [Google Scholar] [CrossRef]
- Wong, A.D.; Ye, M.; Levy, A.F.; Rothstein, J.D.; Bergles, D.E.; Searson, P.C. The Blood-Brain Barrier: An Engineering Perspective. Front. Neuroeng. 2013, 6, 7. [Google Scholar] [CrossRef]
- Smith, Q.R. Transport of Glutamate and Other Amino Acids at the Blood-Brain Barrier. J. Nutr. 2000, 130, 1016S–1022S. [Google Scholar] [CrossRef] [PubMed]
- Zaragozá, R. Transport of Amino Acids Across the Blood-Brain Barrier. Front. Physiol. 2020, 11, 973. [Google Scholar] [CrossRef]
- Yudkoff, M.; Daikhin, Y.; Nissim, I.; Horyn, O.; Luhovyy, B.; Lazarow, A.; Nissim, I. Brain Amino Acid Requirements and Toxicity: The Example of Leucine. J. Nutr. 2005, 135, 1531S–1538S. [Google Scholar] [CrossRef]
- Conway, M.E.; Hutson, S.M. BCAA Metabolism and NH3 Homeostasis. In The Glutamate/GABA-Glutamine Cycle; Schousboe, A., Sonnewald, U., Eds.; Advances in Neurobiology; Springer International Publishing: Cham, Switzerland, 2016; Volume 13, pp. 99–132. ISBN 978-3-319-45094-0. [Google Scholar]
- Griffin, J.W.D.; Bradshaw, P.C. Amino Acid Catabolism in Alzheimer’s Disease Brain: Friend or Foe? Oxidative Med. Cell. Longev. 2017, 2017, 1–15. [Google Scholar] [CrossRef] [PubMed]
- Sperringer, J.E.; Addington, A.; Hutson, S.M. Branched-Chain Amino Acids and Brain Metabolism. Neurochem. Res. 2017, 42, 1697–1709. [Google Scholar] [CrossRef] [PubMed]
- He, W.; Wu, G. Metabolism of Amino Acids in the Brain and Their Roles in Regulating Food Intake. In Amino Acids in Nutrition and Health; Wu, G., Ed.; Advances in Experimental Medicine and Biology; Springer International Publishing: Cham, Switzerland, 2020; Volume 1265, pp. 167–185. ISBN 978-3-030-45327-5. [Google Scholar]
- Salcedo, C.; Andersen, J.V.; Vinten, K.T.; Pinborg, L.H.; Waagepetersen, H.S.; Freude, K.K.; Aldana, B.I. Functional Metabolic Mapping Reveals Highly Active Branched-Chain Amino Acid Metabolism in Human Astrocytes, Which Is Impaired in iPSC-Derived Astrocytes in Alzheimer’s Disease. Front. Aging Neurosci. 2021, 13, 553. [Google Scholar] [CrossRef] [PubMed]
- Suryawan, A.; Hawes, J.W.; Harris, R.A.; Shimomura, Y.; Jenkins, A.E.; Hutson, S.M. A Molecular Model of Human Branched-Chain Amino Acid Metabolism. Am. J. Clin. Nutr. 1998, 68, 72–81. [Google Scholar] [CrossRef] [PubMed]
- Fernstrom, J.D. Branched-Chain Amino Acids and Brain Function. J. Nutr. 2005, 135, 1539S–1546S. [Google Scholar] [CrossRef]
- Bixel, M.G.; Engelmann, J.; Willker, W.; Hamprecht, B.; Leibfritz, D. Metabolism of [U-(13)C]Leucine in Cultured Astroglial Cells. Neurochem. Res. 2004, 29, 2057–2067. [Google Scholar] [CrossRef] [PubMed]
- Murín, R.; Mohammadi, G.; Leibfritz, D.; Hamprecht, B. Glial Metabolism of Isoleucine. Neurochem. Res. 2009, 34, 194–204. [Google Scholar] [CrossRef]
- Murín, R.; Mohammadi, G.; Leibfritz, D.; Hamprecht, B. Glial Metabolism of Valine. Neurochem. Res. 2009, 34, 1195–1203. [Google Scholar] [CrossRef] [PubMed]
- Bixel, M.G.; Hamprecht, B. Generation of Ketone Bodies from Leucine by Cultured Astroglial Cells. J. Neurochem. 1995, 65, 2450–2461. [Google Scholar] [CrossRef]
- Bixel, M.G.; Hamprecht, B. Immunocytochemical Localization of Beta-Methylcrotonyl-CoA Carboxylase in Astroglial Cells and Neurons in Culture. J. Neurochem. 2000, 74, 1059–1067. [Google Scholar] [CrossRef]
- Murín, R.; Verleysdonk, S.; Rapp, M.; Hamprecht, B. Immunocytochemical Localization of 3-Methylcrotonyl-CoA Carboxylase in Cultured Ependymal, Microglial and Oligodendroglial Cells. J. Neurochem. 2006, 97, 1393–1402. [Google Scholar] [CrossRef]
- Murín, R.; Schaer, A.; Kowtharapu, B.S.; Verleysdonk, S.; Hamprecht, B. Expression of 3-Hydroxyisobutyrate Dehydrogenase in Cultured Neural Cells. J. Neurochem. 2008, 105, 1176–1186. [Google Scholar] [CrossRef] [PubMed]
- Garcia-Serrano, A.M.; Duarte, J.M.N. Brain Metabolism Alterations in Type 2 Diabetes: What Did We Learn From Diet-Induced Diabetes Models? Front. Neurosci. 2020, 14, 229. [Google Scholar] [CrossRef] [PubMed]
- Hwang, J.J.; Jiang, L.; Hamza, M.; Rangel, E.S.; Dai, F.; Belfort-DeAguiar, R.; Parikh, L.; Koo, B.B.; Rothman, D.L.; Mason, G.; et al. Blunted Rise in Brain Glucose Levels during Hyperglycemia in Adults with Obesity and T2DM. JCI Insight 2017, 2, e95913. [Google Scholar] [CrossRef]
- Sonneville, R.; den Hertog, H.M.; Güiza, F.; Gunst, J.; Derese, I.; Wouters, P.J.; Brouland, J.-P.; Polito, A.; Gray, F.; Chrétien, F.; et al. Impact of Hyperglycemia on Neuropathological Alterations during Critical Illness. J. Clin. Endocrinol. Metab. 2012, 97, 2113–2123. [Google Scholar] [CrossRef] [PubMed]
- Sickmann, H.M.; Waagepetersen, H.S. Effects of Diabetes on Brain Metabolism—Is Brain Glycogen a Significant Player? Metab. Brain Dis. 2015, 30, 335–343. [Google Scholar] [CrossRef]
- Zhu, H.; Bai, M.; Xie, X.; Wang, J.; Weng, C.; Dai, H.; Chen, J.; Han, F.; Lin, W. Impaired Amino Acid Metabolism and Its Correlation with Diabetic Kidney Disease Progression in Type 2 Diabetes Mellitus. Nutrients 2022, 14, 3345. [Google Scholar] [CrossRef]
- Gondáš, E.; Kráľová Trančíková, A.; Baranovičová, E.; Šofranko, J.; Hatok, J.; Kowtharapu, B.S.; Galanda, T.; Dobrota, D.; Kubatka, P.; Busselberg, D.; et al. Expression of 3-Methylcrotonyl-CoA Carboxylase in Brain Tumors and Capability to Catabolize Leucine by Human Neural Cancer Cells. Cancers 2022, 14, 585. [Google Scholar] [CrossRef] [PubMed]
- Gondáš, E.; Kráľová Trančíková, A.; Šofranko, J.; Majerová, P.; Lučanský, V.; Dohál, M.; Kováč, A.; Murín, R. The Presence of Pyruvate Carboxylase in the Human Brain and Its Role in the Survival of Cultured Human Astrocytes. Physiol. Res. 2023, 72, 403–414. [Google Scholar] [CrossRef]
- Schousboe, A.; Westergaard, N.; Sonnewald, U.; Petersen, S.B.; Huang, R.; Peng, L.; Hertz, L. Glutamate and Glutamine Metabolism and Compartmentation in Astrocytes. Dev. Neurosci. 1993, 15, 359–366. [Google Scholar] [CrossRef]
- Magistretti, P.J. Neuron-Glia Metabolic Coupling and Plasticity. J. Exp. Biol. 2006, 209, 2304–2311. [Google Scholar] [CrossRef] [PubMed]
- Magistretti, P.J.; Sorg, O.; Yu, N.; Martin, J.-L.; Pellerin, L. Neurotransmitters Regulate Energy Metabolism in Astrocytes: Implications for the Metabolic Trafficking between Neural Cells. Dev. Neurosci. 1993, 15, 306–312. [Google Scholar] [CrossRef] [PubMed]
- Magistretti, P.J.; Allaman, I. Lactate in the Brain: From Metabolic End-Product to Signalling Molecule. Nat. Rev. Neurosci. 2018, 19, 235–249. [Google Scholar] [CrossRef]
- Swanson, R.A.; Benington, J.H. Astrocyte Glucose Metabolism under Normal and Pathological Conditions in Vitro. Dev. Neurosci. 1996, 18, 515–521. [Google Scholar] [CrossRef] [PubMed]
- Wiesinger, H.; Hamprecht, B.; Dringen, R. Metabolic Pathways for Glucose in Astrocytes. Glia 1997, 21, 22–34. [Google Scholar] [CrossRef]
- Walz, W.; Mukerji, S. Lactate Production and Release in Cultured Astrocytes. Neurosci. Lett. 1988, 86, 296–300. [Google Scholar] [CrossRef]
- Pellerin, L.; Pellegri, G.; Bittar, P.G.; Charnay, Y.; Bouras, C.; Martin, J.-L.; Stella, N.; Magistretti, P.J. Evidence Supporting the Existence of an Activity-Dependent Astrocyte-Neuron Lactate Shuttle. Dev. Neurosci. 1998, 20, 291–299. [Google Scholar] [CrossRef] [PubMed]
- Horvat, A.; Zorec, R.; Vardjan, N. Lactate as an Astroglial Signal Augmenting Aerobic Glycolysis and Lipid Metabolism. Front. Physiol. 2021, 12, 735532. [Google Scholar] [CrossRef] [PubMed]
- Schousboe, A.; Scafidi, S.; Bak, L.K.; Waagepetersen, H.S.; McKenna, M.C. Glutamate Metabolism in the Brain Focusing on Astrocytes. In Glutamate and ATP at the Interface of Metabolism and Signaling in the Brain; Parpura, V., Schousboe, A., Verkhratsky, A., Eds.; Advances in Neurobiology; Springer International Publishing: Cham, Switzerland, 2014; Volume 11, pp. 13–30. ISBN 978-3-319-08893-8. [Google Scholar]
- Rabah, Y.; Francés, R.; Minatchy, J.; Guédon, L.; Desnous, C.; Plaçais, P.-Y.; Preat, T. Glycolysis-Derived Alanine from Glia Fuels Neuronal Mitochondria for Memory in Drosophila. Nat. Metab. 2023, 5, 2002–2019. [Google Scholar] [CrossRef]
- Schousboe, A.; Sonnewald, U.; Waagepetersen, H.S. Differential Roles of Alanine in GABAergic and Glutamatergic Neurons. Neurochem. Int. 2003, 43, 311–315. [Google Scholar] [CrossRef]
- Kumar, A.; Bachhawat, A.K. Pyroglutamic Acid: Throwing Light on a Lightly Studied Metabolite. Curr. Sci. 2012, 102, 288–297. [Google Scholar]
- Jensen, N.J.; Wodschow, H.Z.; Nilsson, M.; Rungby, J. Effects of Ketone Bodies on Brain Metabolism and Function in Neurodegenerative Diseases. Int. J. Mol. Sci. 2020, 21, 8767. [Google Scholar] [CrossRef]
- García-Rodríguez, D.; Giménez-Cassina, A. Ketone Bodies in the Brain Beyond Fuel Metabolism: From Excitability to Gene Expression and Cell Signaling. Front. Mol. Neurosci. 2021, 14, 732120. [Google Scholar] [CrossRef] [PubMed]
- Buser, D.P.; Ritz, M.-F.; Moes, S.; Tostado, C.; Frank, S.; Spiess, M.; Mariani, L.; Jenö, P.; Boulay, J.-L.; Hutter, G. Quantitative Proteomics Reveals Reduction of Endocytic Machinery Components in Gliomas. EBioMedicine 2019, 46, 32–41. [Google Scholar] [CrossRef] [PubMed]
- Pavel, M.; Renna, M.; Park, S.J.; Menzies, F.M.; Ricketts, T.; Füllgrabe, J.; Ashkenazi, A.; Frake, R.A.; Lombarte, A.C.; Bento, C.F.; et al. Contact Inhibition Controls Cell Survival and Proliferation via YAP/TAZ-Autophagy Axis. Nat. Commun. 2018, 9, 2961. [Google Scholar] [CrossRef] [PubMed]
Parameter (Unit) | Glucose | |
---|---|---|
5 mM | 25 mM | |
Protein content (mg) | 0.11 ± 0.03 | 0.09 ± 0.03 |
Cell survival (%) | 70 ± 5 | 67 ± 6 |
as [LDH] (U/g) | 266 ± 29 | 231 ± 50 |
Concentration (µM) | Medium | Glucose | |
---|---|---|---|
5 mM | 25 mM | ||
lactate | 801 ± 38 | 2195 ± 176 ** | 1862 ± 60 |
pyruvate | 917 ± 26 | 341 ± 27 ** | 430 ± 27 |
citrate | 18.5 ± 0.5 | 25 ± 1 | 26 ± 1 |
acetate | 90 ± 12 | 136 ± 14 | 128 ± 40 |
acetone | 29 ± 2 | 174 ± 78 | 253 ± 98 |
3-hydroxybutyrate | 0 | 56 ± 2 | 62 ± 5 |
glutamine | 5045 ± 180 | 3878 ± 136 * | 3716 ± 50 |
leucine | 793 ± 15 | 714 ± 18 | 727 ± 7 |
α-ketoisocaproate | 22 ± 4 | 61 ± 5 ** | 45 ± 3 |
isoleucine | 751 ± 19 | 663 ± 24 | 674 ± 12 |
α-keto-methylvalerate | 14 ± 2 | 53 ± 4 *** | 39 ± 2 |
valine | 773 ± 10 | 745 ± 14 | 748 ± 16 |
α-ketoisovalerate | 7 ± 1 | 48 ± 4 ** | 35 ± 2 |
alanine | 148 ± 2 | 206 ± 6 ** | 190 ± 3 |
histidine | 464 ± 8 | 467 ± 12 | 465 ± 11 |
lysine | 867 ± 40 | 1024 ± 21 * | 980 ± 68 |
methionine | 209 ± 3 | 222 ± 5 ** | 210 ± 2 |
phenylalanine | 470 ± 6 | 487 ± 8 | 482 ± 8 |
threonine | 1016 ± 34 | 1043 ± 41 | 1014 ± 41 |
tryptophan | 21 ± 1 | 23 ± 2 | 23 ± 1 |
tyrosine | 367 ± 6 | 381 ± 7 * | 371 ± 6 |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Gondáš, E.; Baranovičová, E.; Šofranko, J.; Murín, R. Hyperglycemia Stimulates the Irreversible Catabolism of Branched-Chain Amino Acids and Generation of Ketone Bodies by Cultured Human Astrocytes. Biomedicines 2024, 12, 1803. https://doi.org/10.3390/biomedicines12081803
Gondáš E, Baranovičová E, Šofranko J, Murín R. Hyperglycemia Stimulates the Irreversible Catabolism of Branched-Chain Amino Acids and Generation of Ketone Bodies by Cultured Human Astrocytes. Biomedicines. 2024; 12(8):1803. https://doi.org/10.3390/biomedicines12081803
Chicago/Turabian StyleGondáš, Eduard, Eva Baranovičová, Jakub Šofranko, and Radovan Murín. 2024. "Hyperglycemia Stimulates the Irreversible Catabolism of Branched-Chain Amino Acids and Generation of Ketone Bodies by Cultured Human Astrocytes" Biomedicines 12, no. 8: 1803. https://doi.org/10.3390/biomedicines12081803