Quipazine treatment exacerbates oxidative stress in glutamate-induced HT-22 neuronal cells

Abstract

Objectives: Quipazine is a serotonin agonist. It is known that serotonin, an important neurotransmitter, contributes to the etiology of psychiatric and many neurodegenerative diseases. However, the effect of the serotonin agonist quipazine on HT-22 cells in glutamate-induced cytotoxicity is unknown. This study aims to investigate the effect of quipazine on increased oxidative stress (OS) as a result of glutamate-induced cytotoxicity in HT-22 cells.

Methods: The cells were divided into 4 groups, Control group: no treatment was applied, Glutamate group: glutamate was incubated at 10 mM for 24 h, Quipazine group: incubated with different doses of quipazine for 24 h, Quipazine+Glutamate group were pre-treated with various concentrations (25, 50, 100 and 200 µM) of quipazine for 1 h and then exposed to 10 mM glutamate for 24 h. Cell viability rate between groups was measured by the XTT assay. OS and antioxidant levels were measured with the Total Oxidant Status (TOS) and Total Antioxidant Status (TAS) Elisa kits, and Caspase-3 levels were also examined in caspase activity. 

Results: Quipazine at different concentrations showed significant differences in cell viability in HT-22 cells. An appropriate dose of 25 µM was accepted for quipazine in the study. Quipazine treatment with glutamate-toxicity in the cells further reduced TAS levels and significantly increased TOS levels. It was also observed that the Caspase-3 level increased more in the Quipazine + Glutamate group according to the Glutamate group. 

Conclusions: The results determined that the use of quipazine is an agent that will further increase the neurodegeneration caused by glutamate toxicity. 

Keywords

• Quipazine
• glutamate-toxicity
• oxidative stress
• HT-22 cell
• cell viability

References

1. 1. Dong XX, Wang Y, Qin ZH. Molecular mechanisms of excitotoxicity and their relevance to pathogenesis of neurodegenerative diseases. Acta Pharmacol Sin 2009;30:379-87.
2. 2. Olney J W. Brain lesions, obesity, and other disturbances in mice treated with monosodium glutamate. Science 1969;164:719-21.
3. 3. Barnes GN, Slevin JT. Ionotropic glutamate receptor biology: effect on synaptic connectivity and function in neurological disease. Curr Med Chem 2003;10:2059-72.
4. 4. Danbolt NC. Glutamate uptake. Prog Neurobiol 2001;65:1-105.
5. 5. Pereira CF, de Oliveira CR. Oxidative glutamate toxicity involves mitochondrial dysfunction and perturbation of intracellular Ca2+ homeostasis. Neurosci Res 2000;37:227-36.
6. 6. Wang W, Zhang F, Li L, Tang F, Siedlak SL, Fujioka H, et al. MFN2 couples glutamate excitotoxicity and mitochondrial dysfunction in motor neurons. J Biol Chem 2015;290:168-82.
7. 7. Atlante A, Calissano P, Bobba A, Giannattasio S, Marra E, Passarella S. Glutamate neurotoxicity, oxidative stress and mitochondria. FEBS Lett 2001;497:1-5.
8. 8. Lewerenz J, Maher P. Chronic glutamate toxicity in neurodegenerative diseases – what is the evidence? Front Neurosci 2015;9:469.

9. 9. Lancelot E, Beal MF. Glutamate toxicity in chronic neurodegenerative disease. Prog Brain Res 1998;116:331-47.
10. 10. Kritis AA, Stamoula EG, Paniskaki KA, Vavilis TD. Researching glutamate-induced cytotoxicity in different cell lines: a comparative/collective analysis/study. Front Cell Neurosci 2015;9:91.
11. 11. Swann-Thomsen HE, Viall DD, Brumley MR. Acute intrathecal administration of quipazine elicits air-stepping behavior. Behav Pharmacol 2021;32:259-64.
12. 12. Liang H, Liu N, Wang R, Zhang Y, Chen J, Dai Z, et al. N-acetyl serotonin alleviates oxidative damage by activating nuclear factor erythroid 2-related factor 2 signaling in porcine enterocytes. Antioxidants (Basel) 2020;9:303.
13. 13. Nocito A, Dahm F, Jochum W, Jang JH, Georgiev P, Bader M, et al. Serotonin mediates oxidative stress and mitochondrial toxicity in a murine model of nonalcoholic steatohepatitis. Gastroenterology 2007;133:608-18.
14. 14. Vasicek O, Lojek A, Ciz M. Serotonin and its metabolites reduce oxidative stress in murine RAW264.7 macrophages and prevent inflammation. J Physiol Biochem 2020;76:49-60.
15. 15. Banerjee E, Nandagopal K. Does serotonin deficit mediate susceptibility to ADHD? Neurochem Int 2015;82:52-68.
16. 16. Goodwin GM, Green AR. A behavioural and biochemical study in mice and rats of putative selective agonists and antagonists for 5-HT1 and 5-HT2 receptors. Br J Pharmacol 1985;84:743-53.
17. 17. Round A, Wallis D. Further studies on the blockade of 5-HT depolarizations of rabbit vagal afferent and sympathetic ganglion cells by MDL 72222 and other antagonists. Neuropharmacol 1987;26:39-48.
18. 18. Davis JB, Maher P. Protein kinase C activation inhibits glutamate-induced cytotoxicity in a neuronal cell line. Brain Res 1994;652:169-73.
19. 19. Taskiran AS, Ergul M. The effect of salmon calcitonin against glutamate-induced cytotoxicity in the C6 cell line and the roles the inflammatory and nitric oxide pathways play. Metab Brain Dis 2021;36:1985-93.
20. 20. Kennaway DJ, Rowe S, Ferguson S. Serotonin agonists mimic the phase shifting effects of light on the melatonin rhythm in rats. Brain Res 1996;737:301-7.
21. 21. Kohler M, Kalkowski A, Wollnik F. Serotonin agonist quipazine induces photic-like phase shifts of the circadian activity rhythm and c-Fos expression in the rat suprachiasmatic nucleus. J Biol Rhythms 1999;14:131-40.
22. 22. Shinagawa S. Serotonin protects C6 glioma cells from glutamate toxicity. Neurosci 1994;59:1043-50.
23. 23. Menezes AC, Carvalheiro M, Ferreira de Oliveira JMP, Ascenso A, Oliveira H. Cytotoxic effect of the serotonergic drug 1-(1-Naphthyl)piperazine against melanoma cells. Toxicol In Vitro 2018;47:72-8.
24. 24. Yoo JM, Lee BD, Lee SJ, Ma JY, Kim MR. Anti-apoptotic effect of N-palmitoyl serotonin on glutamate-mediated apoptosis through secretion of BDNF and activation of TrkB/CREB pathway in HT-22 cells. Eur J Lipid Sci Technol 2018;120:1700397.
25. 25. Doğan M, Yildizhan K. Investigation of the effect of paracetamol against glutamate-induced cytotoxicity in C6 glia cells. Cumhuriyet Sci J 2021:42;789-94.
26. 26. Wang J, Wang F, Mai D, Qu S. Molecular mechanisms of glutamate toxicity in parkinson's disease. Front Neurosci 2020;14:585584.
27. 27. Jiang T, Cheng H, Su J, Wang X, Wang Q, Chu J, et al. Gastrodin protects against glutamate-induced ferroptosis in HT-22 cells through Nrf2/HO-1 signaling pathway. Toxicol In Vitro 2020;62:104715.
28. 28. Sillapachaiyaporn C, Rangsinth P, Nilkhet S, Ung AT, Chuchawankul S, Tencomnao T. Neuroprotective effects against glutamate-induced HT-22 hippocampal cell damage and caenorhabditis elegans lifespan/healthspan enhancing activity of auricularia polytricha mushroom extracts. Pharmaceuticals (Basel) 2021;14:1001.
29. 29. Sabogal-Guaqueta AM, Hobbie F, Keerthi A, Oun A, Kortholt A, Boddeke E, et al. Linalool attenuates oxidative stress and mitochondrial dysfunction mediated by glutamate and NMDA toxicity. Biomed Pharmacother 2019;118:109295.
30. 30. Annunziato L, Cataldi M, Pignataro G, Secondo A, Molinaro P. Glutamate-independent calcium toxicity: introduction. Stroke 2007;38(2 Suppl):661-4.
31. 31. Lee HJ, Spandidos DA, Tsatsakis A, Margina D, Izotov BN, Yang SH. Neuroprotective effects of Scrophularia buergeriana extract against glutamate-induced toxicity in SH-SY5Y cells. Int J Mol Med 2019;43:2144-52.
32. 32. Tonsomboon A, Prasanth MI, Plaingam W, Tencomnao T. Kaempferia parviflora rhizome extract inhibits glutamate-induced toxicity in HT-22 mouse hippocampal neuronal cells and extends longevity in Caenorhabditis elegans. Biology (Basel) 2021;10:264.
33. 33. Lau A, Tymianski M. Glutamate receptors, neurotoxicity and neurodegeneration. Pflugers Arch 2010;460:525-42.
34. 34. Hynd MR, Scott HL, Dodd PR. Glutamate-mediated excitotoxicity and neurodegeneration in Alzheimer's disease. Neurochem Int 2004;45:583-95.
35. 35. Gulcin I. Measurement of antioxidant ability of melatonin and serotonin by the DMPD and CUPRAC methods as trolox equivalent. J Enzyme Inhib Med Chem 2008;23:871-6.