Protective Effect Of Low Dose Of Methamphetamine On The Amount Of Extracellular Glutamine In Primary Fetal Human Astrocytes Induced By Amyloid Beta

Document Type : Original Article

Authors

1 Department of Biology, College of Science, Damghan Branch, Islamic Azad University, Damghan, Iran

2 Department of Biology, Faculty of Science, University of Zanjan, Zanjan, Iran

3 Laboratory of Neuro-Organic Chemistry, Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran

Abstract

Objective: Change in astrocytes is one of the first pathological symptoms of Alzheimer’s disease (AD). Understanding
the signaling pathways in astrocytes can be a great help in treating of AD. This study aimed to investigate signaling pathway relations between low dose of methamphetamine (METH), the apoptosis, cell cycle, and glutamine (Gln) pathways in the activated astrocyte.
Materials and Methods: In this experimental study, the activated astrocyte cells were exposed to a low dose of METH
(12.5 µM) which was determined by Thiazolyl blue tetrazolium bromide (MTT) method. The groups were: group 1 cells with Aβ, group 2 cells with METH, group 3 cells with METH after 24 hours of adding Aβ (Aβ+METH, treated group), group 4 cells with Aβ after 24 hours of adding METH (METH+Aβ, prevention group), and group 5 as the control. The Gln was assayed by high-performance liquid chromatography (HPLC), and also the apoptosis, and cell cycle and BAX, BCL-X expression was evaluated.
Results: The amount of Gln was increased, and the value of late and early apoptosis was reduced in the treatment groups, and necrosis is decreased in the prevention group (group 4 compared to group 1). Moreover, it was revealed through cell cycle analysis that G2 in group 4 was reduced compared to group 1 and the expression of BAX, BAX/ BCL-X, and BCL-X in group 3 and group 4, was decreased and increased, respectively compared to group 1.
Conclusion: These findings suggest that perhaps a non-toxic dosage of METH (low dose) can reduce the amount of apoptosis and BAX expression and increase the expression of BCL-X. Furthermore, the cells are arrested in the G2 phase and can raise the amount of extracellular glutamine, which has a protective role in neuron cells. These findings
may provide a new perspective to design a new drug with less toxic results. 

Keywords


  1. Sofroniew MV, Vinters HV. Astrocytes: biology and pathology. Acta Neuropathol. 2010; 119(1): 7-35.
  2. Back A, Tupper KY, Bai T, Chiranand P, Goldenberg FD, Frank JI, et al. Ammonia-induced brain swelling and neurotoxicity in an organotypic slice model. Neurol Res. 2011; 33(10): 1100-1108.
  3. Du J, Song D, Li Y, Liu J, Huang X, Li B, et al. Saikosaponin-D mitigates oxidation in SH-SY5Y cells stimulated by glutamate through activation of Nrf2 pathway: involvement of PI3K. Neurotox Res. 2022; 40(1): 230-240.
  4. Rau TF, Kothiwal AS, Rova AR, Brooks DM, Rhoderick JF, Poulsen AJ, et al. Administration of low dose methamphetamine 12 h after a severe traumatic brain injury prevents neurological dysfunction and cognitive impairment in rats. Exp Neurol. 2014; 253: 31-40.
  5. Liddelow SA, Barres BA. Reactive astrocytes: production, function, and therapeutic potential. Immunity. 2017; 46(6): 957-967.
  6. Olabarria M, Noristani HN, Verkhratsky A, Rodríguez JJ. Agedependent decrease in glutamine synthetase expression in the hippocampal astroglia of the triple transgenic Alzheimer’s disease mouse model: mechanism for deficient glutamatergic transmission? Mol Neurodegener. 2011; 6: 55.
  7. Han X, Zhang T, Liu H, Mi Y, Gou X. Astrocyte senescence and Alzheimer’s disease: a review. Front Aging Neurosci. 2020; 12: 148.
  8. Pajarillo E, Rizor A, Lee J, Aschner M, Lee E. The role of astrocytic glutamate transporters GLT-1 and GLAST in neurological disorders: Potential targets for neurotherapeutics. Neuropharmacology. 2019; 161: 107559.
  9. Bellaver B, Souza DG, Souza DO, Quincozes-Santos A. Hippocampal astrocyte cultures from adult and aged rats reproduce changes in glial functionality observed in the aging brain. Mol Neurobiol. 2017; 54(4): 2969-2985.
  10. Acosta C, Anderson HD, Anderson CM. Astrocyte dysfunction in Alzheimer disease. J Neurosci Res. 2017; 95(12): 2430-2447.
  11. Howland DS, Liu J, She Y, Goad B, Maragakis NJ, Kim B, et al. Focal loss of the glutamate transporter EAAT2 in a transgenic rat model of SOD1 mutant-mediated amyotrophic lateral sclerosis (ALS). Proc Natl Acad Sci USA. 2002; 99(3): 1604-1609.
  12. Chen J, Herrup K. Glutamine acts as a neuroprotectant against DNA damage, beta-amyloid and H2O2-induced stress. PLoS One. 2012; 7(3): e33177.
  13. Borgan FR, Jauhar S, McCutcheon RA, Pepper FS, Rogdaki M, Lythgoe DJ, et al. Glutamate levels in the anterior cingulate cortex in un-medicated first episode psychosis: a proton magnetic resonance spectroscopy study. Sci Rep. 2019; 9(1): 8685.
  14. Yao M, Nguyen TV, Pike CJ. Beta-amyloid-induced neuronal apoptosis involves c-Jun N-terminal kinase-dependent downregulation of Bcl-w. J Neurosci. 2005; 25(5): 1149-1158.
  15. Garwood CJ, Pooler AM, Atherton J, Hanger DP, Noble W. Astrocytes are important mediators of Aβ-induced neurotoxicity and tau phosphorylation in primary culture. Cell Death Dis. 2011; 2(6): e167.
  16. Jana A, Pahan K. Fibrillar amyloid-beta-activated human astroglia kill primary human neurons via neutral sphingomyelinase: implications for Alzheimer’s disease. J Neurosci. 2010; 30(38): 12676- 12689.
  17. Prakash MD, Tangalakis K, Antonipillai J, Stojanovska L, Nurgali K, Apostolopoulos V. Methamphetamine: effects on the brain, gut and immune system. Pharmacol Res. 2017; 120: 60-67.
  18. Ghahghaei A, Bathaie SZ, Kheirkhah H, Bahraminejad E. The protective effect of crocin on the amyloid fibril formation of Aβ42 peptide in vitro. Cell Mol Biol Lett. 2013; 18(3): 328-339.
  19. Rau T, Ziemniak J, Poulsen D. The neuroprotective potential of low-dose methamphetamine in preclinical models of stroke and traumatic brain injury. Prog Neuropsychopharmacol Biol Psychiatry. 2016; 64: 231-236.
  20. Rau TF, Kothiwal AS, Rova AR, Brooks DM, Poulsen DJ. Treatment with low-dose methamphetamine improves behavioral and cognitive function after severe traumatic brain injury. J Trauma Acute Care Surg. 2012; 73(2 Suppl 1): S165-172.
  21. Thrash B, Karuppagounder SS, Uthayathas S, Suppiramaniam V, Dhanasekaran M. Neurotoxic effects of methamphetamine. Neurochem Res. 2010; 35(1): 171-179.
  22. Charbonneau JR, Furtak T, Lefebvre J, Gauthier ER. Bcl-xL expression interferes with the effects of L-glutamine supplementation on hybridoma cultures. Biotechnol Bioeng. 2003; 81(3): 279-290.
  23. Miguel-Hidalgo JJ. The role of glial cells in drug abuse. Curr Drug Abuse Rev. 2009; 2(1): 76-82.
  24. Matos M, Augusto E, Oliveira CR, Agostinho P. Amyloid-beta peptide decreases glutamate uptake in cultured astrocytes: involvement of oxidative stress and mitogen-activated protein kinase cascades. Neuroscience. 2008; 156(4): 898-910.
  25. Sharif A, Prevot V. Isolation and culture of human astrocytes. Methods Mol Biol. 2012; 814: 137-151.
  26. White JA, Manelli AM, Holmberg KH, Van Eldik LJ, Ladu MJ. Differential effects of oligomeric and fibrillar amyloid-beta 1-42 on astrocyte- mediated inflammation. Neurobiol Dis. 2005; 18(3): 459-465.
  27. Bahuguna A, Khan I, Bajpai V K,Kang S C. MTT assay to evaluate the cytotoxic potential of a drug. Bangladesh Journal of Pharmacology. 2017; 12(2).
  28. Shakibaie M, Vaezjalali M, Rafii-Tabar H, Sasanpour P. Phototherapy alters the oncogenic metabolic activity of breast cancer cells. Photodiagnosis Photodyn Ther. 2020; 30: 101695.
  29. Riccardi C, Nicoletti I. Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc. 2006; 1(3): 1458-1461.
  30. Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008; 3(6): 1101-1108.
  31. Antuono PG, Jones JL, Wang Y, Li SJ. Decreased glutamate + glutamine in Alzheimer’s disease detected in vivo with (1)H-MRS at 0.5 T. Neurology. 2001; 56(6): 737-742.
  32. Rau TF, Kothiwal A, Zhang L, Ulatowski S, Jacobson S, Brooks DM, et al. Low dose methamphetamine mediates neuroprotection through a PI3K-AKT pathway. Neuropharmacology. 2011; 61(4): 677-686.
  33. El Ayadi A, Zigmond MJ. Low concentrations of methamphetamine can protect dopaminergic cells against a larger oxidative stress injury: mechanistic study. PLoS One. 2011; 6(10): e24722.
  34. Baghbaderani S, Hashemi M, Ebrahimi-Ghiri M, Zarrindast MR, Nasehi M, Entezari M. Curcumin prevents cognitive deficits in the bile duct ligated rats. Psychopharmacology (Berl). 2020; 237(12): 3529-3537.
  35. Currais A, Hortobágyi T, Soriano S. The neuronal cell cycle as a mechanism of pathogenesis in Alzheimer’s disease. Aging (Albany NY). 2009; 1(4): 363-371.
  36. Bajić VP, Su B, Lee HG, Kudo W, Siedlak SL, Zivković L, et al. Mislocalization of CDK11/PITSLRE, a regulator of the G2/M phase of the cell cycle, in Alzheimer disease. Cell Mol Biol Lett. 2011; 16(3): 359-372.
  37. Chen SD, Yang JL, Lin YC, Chao AC, Yang DI. Emerging roles of inhibitor of differentiation-1 in Alzheimer’s disease: cell cycle reentry and beyond. Cells. 2020; 9(7): 1746.
  38. Bennecib M, Gong CX, Grundke-Iqbal I, Iqbal K. Role of protein phosphatase-2A and -1 in the regulation of GSK-3, cdk5 and cdc2 and the phosphorylation of tau in rat forebrain. FEBS Lett. 2000; 485(1): 87-93.
  39. Willette AA, Bendlin BB, Starks EJ, Birdsill AC, Johnson SC, Christian BT, et al. Association of insulin resistance with cerebral glucose uptake in late middle-aged adults at risk for alzheimer disease. JAMA Neurol. 2015; 72(9): 1013-1020.
  40. Abdul Muneer PM, Alikunju S, Szlachetka AM, Haorah J. Methamphetamine inhibits the glucose uptake by human neurons and astrocytes: stabilization by acetyl-L-carnitine. PLoS One. 2011; 6(4): e19258.