Postmortem Study of Molecular and Histological Changes in the CA1 Hippocampal Region of Chronic Methamphetamine User

Document Type : Research article

Authors

1 Hearing Disorders Research Center, Loghman Hakim Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

2 Laser Application in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

3 Proteomics Research Center, Faculty of Paramedicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

4 Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

5 Cellular and Molecular Research Center, Faculty of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.

Abstract

Methamphetamine (Meth) is recognized as one of the most important new distributed
abused drug that causes severe damage to the different parts of the brain, especially
hippocampus. Previous studies have demonstrated that Meth can induce apoptosis and cell
death in the brain. In this study, we evaluated the long-term effects of Meth abuse in the
CA1 region of postmortem hippocampus. Postmortem molecular and histological analysis
was performed for five non-addicted subjects and five Meth addicted ones. Iba-1 (microglia)
and glial fibrillary acidic protein, GFAP (astrocytes) expression were assayed by western
blotting and immunohistochemistry (IHC) methods. Histopathological assessment was done
with stereological counts of hippocampal cells stained with hematoxylin and eosin (H and E).
Tunel staining was used to detect DNA damage in human brains. In addition, protein-protein
interaction analysis network was investigated. Western blotting and immunohistochemistry
assay showed overexpression of GFAP and Iba-1 protein in the CA1 hippocampal region
of Meth users’ brain. Stereological analysis in the CA1 region revealed increased neuron
degeneration. Furthermore, significant apoptosis and cell death were confirmed by Tunel assay
in the hippocampus. The prominent role of TLR4, IL1B, CASP1, and NLRP3 in the molecular
mechanism of Meth was highlighted via PPI network analysis. Chronic Meth use can induce
GFAP and Iba-1 upregulation and neuronal apoptosis in the CA1 region of the postmortem
hippocampus.

Keywords

Main Subjects


References
Urbina A and Jones K. Crystal methamphetamine, its
analogues, and HIV infection: medical and psychiatric
aspects of a new epidemic. Clin. Infect. Dis. (2004)
38: 890–4.
Freese TE, Obert J, Dickow A, Cohen J and Lord
RH. Methamphetamine abuse: issues for special
populations. J. Psychoactive Drugs (2000) 32: 177–82.
Snider SE, Vunck SA, van den Oord EJ, Adkins
DE, McClay JL and Beardsley PM. The glial cell
modulators, ibudilast and its amino analog, AV1013,
attenuate methamphetamine locomotor activity and
its sensitization in mice. Eur. J. Pharmacol. (2012)
679: 75–80.
Krasnova IN, Justinova Z, Ladenheim B, Jayanthi S,
McCoy MT, Barnes C, Warner JE, Goldberg SR and
(1)
(2)
(3)
(4)
Postmortem Study of Molecular and Histological Changes in the Hippocampus
2081
Cadet JL. Methamphetamine self-administration is
associated with persistent biochemical alterations in
striatal and cortical dopaminergic terminals in the rat.
PLoS One (2010) 5: e8790.
North A, Swant J, Salvatore MF, Gamble-George
J, Prins P, Butler B, Mittal MK, Heltsley R, Clark
JT and Khoshbouei H. Chronic methamphetamine
exposure produces a delayed, long-lasting memory
deficit. Synapse (2013) 67: 245-57.
Souza DO, dos Santos Sales V, de Souza Rodrigues
CK, de Oliveira LR, Lemos IC, de Araújo Delmondes
G, Monteiro ÁB and do Nascimento EP. Phytochemical
analysis and central effects of Annona muricata
Linnaeus: possible involvement of the gabaergic and
monoaminergic systems. Iran. J. Pharm. Res. (2018)
17: 1306-17.
Zahari Z, Lee CS, Ibrahim MA, Musa N, Yasin
MA, Lee YY, Tan SC, Mohamad N and Ismail R.
Relationship between serum methadone concentration
and cold pressor pain sensitivity in patients undergoing
methadone maintenance therapy. Iran. J. Pharm. Res.
(2018) 17 (Suppl): 8-16.
Hanson JE, Birdsall E, Seferian KS, Crosby MA,
Keefe KA, Gibb JW, Hanson GR and Fleckenstein
AE. Methamphetamine-induced dopaminergic deficits
and refractoriness to subsequent treatment. Eur. J.
Pharmacol. (2009) 607: 68-73.
Kuhn DM, Francescutti-Verbeem DM and Thomas
DM. Dopamine disposition in the presynaptic process
regulates the severity of methamphetamine-induced
neurotoxicity. Ann. NY. Acad. Sci. (2008) 1139: 118.
Cadet JL and Bisagno V. Glial-neuronal ensembles:
partners in drug addiction-associated synaptic
plasticity. Front. Pharmacol. (2014) 5: 204.
Cappon GD, Morford LL and Vorhees CV. Ontogeny
of methamphetamine-Induced neurotoxicity and
associated hyperthermic response. Brain Res. Dev.
(1997) 103: 155–62.
Fukumura M, Cappon GD, Pu C, Broening HW and
Vorhees CV. A single dose model of methamphetamineinduced neurotoxicity in rats: effects on neostriatal
monoamines and glial fibrillary acidic protein. Brain
Res. (1998) 806: 1-7.
Pu C and Vorhees CV. Developmental dissociation of
methamphetamine-induced depletion of dopaminergic
terminals and astrocyte reaction in rat striatum. Brain
Res. Dev. (1993) 72: 325–8.
O′Callaghan JP and Miller DB. Neurotoxicity profiles
of substituted amphetamines in the C57BL/6J mouse.
J. Pharmacol. Exp. Ther. (1994) 270: 741-51.
Xu W, Zhu JP and Angulo JA. Induction of striatal
pre- and postsynaptic damage by methamphetamine
requires the dopamine receptors. Synapse (2005) 58:
110–21.
Aschner M, Guilarte TR, Schneider JS and Zheng
W. Manganese: recent advances in understanding its
transport and neurotoxicity. Toxicol. Appl. Pharmacol.
(2007) 221: 131–47.
Pu C and Vorhees CV. Protective effects of MK-801 on
methamphetamine-induced depletion of dopaminergic
and serotonergic terminals and striatal astrocytic
response: An immunohistochemical study. Synapse
(1995) 19: 97-104.
Fujita Y, Kunitachi S, Iyo M and Hashimoto
K. The antibiotic minocycline prevents
methamphetamineinduced rewarding effects in mice.
Pharmacol. Biochem. Behav. (2012) 101: 303–6.
Narita M, Miyatake M, Shibasaki M, Shindo K,
Nakamura A, Kuzumaki N, Nagumo Y and Suzuki
T. Direct evidence of astrocytic modulation in the
development of rewarding effects induced by drugs of
abuse. Neuropsychopharmacology (2006) 31: 2476–
88.
Zhang L, Kitaichi K, Fujimoto Y, Nakayama H, Shimizu
E, Iyo M and Hashimoto K. Protective effects of
minocycline on behavioral changes and neurotoxicity
in mice after administration of methamphetamine.
Prog. Neuropsychopharmacol. Biol. Psychiatry (2006)
30: 1381–93.
Snider SE, Vunck SA, van den Oord EJ, Adkins
DE, McClay JL and Beardsley PM. The glial cell
modulators, ibudilast and it′s amino analog, AV1013,
attenuate Methamphetamine locomotor activity and
its sensitization in mice. Eur. J. Pharmacol. (2012)
679: 75-80.
Sriram K, Miller DB and O′Callaghan JP. Minocycline
attenuates microglial activation but fails to mitigate
striatal dopaminergic neurotoxicity: role of tumor
necrosis factor-alpha. J. Neurochem. (2006) 96: 706–
18.
Chen H, Uz T and Manev H. Minocycline affects
cocaine sensitization in mice. Neurosci. Lett. (2009)
452: 258–61.
Saadati F, Mahdikia H, Abbaszadeh HA, Abdollahifar
MA, Khoramgah MS and Shokri B.Comparison of
direct and indirect cold atmospheric-pressure plasma
methods in the B16F10 melanoma cancer cells
treatment. Sci. Rep. (2018) 8: 7689.
Darabi S, Tiraihi T, Delshad A, Sadeghizadeh M,
Khalil W and Taheri T. In-vitro non-viral murine proneurotrophin 3 gene transfer into rat bone marrow
stromal cells. J. Neurol. Sci. (2017) 375: 137-45.
Noorafshan A, Asadi-Golshan R, Abdollahifar MA
and Karbalay-Doust S. Protective role of curcumin
against sulfite-induced structural changes in rats′
medial prefrontal cortex. Nutr. Neurosci. (2015) 18:
248-55.
Noorafshan A, Abdollahifar MA and KarbalayDoust S. Stress changes the spatial arrangement of
neurons and glial cells of medial prefrontal cortex and
sertraline and curcumin prevent it. Psychiatry Investig.
(2015) 12: 73-80.
Sofroniew MV and Vinters HV. Astrocytes: biology
and pathology. Acta Neuropathol. (2010) 119: 7-35.
Yang X, Wang Y, Li Q, Zhong Y and Chen L. The main
molecular mechanisms underlying methamphetamineinduced neurotoxicity and implications for
pharmacological treatment. Front. Mol. Neurosci.
(5)
(6)
(7)
(8)
(9)
(10)
(11)
(12)
(13)
(14)
(15)
(16)
(17)
(18)
(19)
(20)
(21)
(22)
(23)
(24)
(25)
(26)
(27)
(28)
(29)
 Mahmoudiasl Ghr et al. / IJPR (2019), 18 (4): 2067-2082
2082
(2018) 11: 186.
Anezaki T, Ishiguro H, Hozumi I, Inuzuka T, Hiraiwa
M, Kobayashi H, Yuguchi T, Wanaka A, Uda Y and
Miyatake T. Expression of growth inhibitory factor
(GIF) in normal and injured rat brains. Neurochem.
Int. (1995) 27: 89-94.
Abbott NJ, Rönnbäck L and Hansson E. Astrocyteendothelial interactions at the blood-brain barrier. Nat.
Rev. Neurosci. (2006) 7: 41-53.
Goldstein GW. Endothelial cell-astrocyte interactions.
A cellular model of the blood-brain barrier. Ann. NY.
Acad. Sci. (1988) 529: 31-9.
Sharma HS and Kiyatkin EA. Rapid morphological
brain abnormalities during acute methamphetamine
intoxication in the rat: an experimental study using
light and electron microscopy. J. Chem. Neuroanat.
(2009) 37: 18-32.
Bowyer JF and Joseph PH. Amphetamine and
methamphetamine-induced hyperthermia: Implications
of the effects produced in brain vasculature and
peripheral organs to forebrain neurotoxicity.
Temperature (Austin) (2014) 1: 172–82.
Almalki AH, Das SC, Alshehri FS, Althobaiti YS
and Sari Y. Effects of sequential ethanol exposure
and repeated high-dose methamphetamine on striatal
and hippocampal dopamine, serotonin and glutamate
tissue content in Wistar rats. Neurosci. Lett. (2018)
665: 61-6.
Shih SC, Prag G, Smitha AF, Myra AS, James HH
and Linda H. A ubiquitin-binding motif required for
intramolecular monoubiquitylation, the CUE domain.
EMBO J. (2003) 22: 1273–81.
Chen RZ, Akbarian S, Tudor M and Jaenisch R.
Deficiency of methyl-CpG binding protein-2 in CNS
neurons results in a Rett-like phenotype in mice. Nat.
Genet. (2001) 27: 327-31.
Jirapa C, Sujira M, Rachneekorn S, Piyarat G and
Banthit C. Role of melatonin in reducing amphetamineinduced degeneration in substantia nigra of rats via
calpain and calpastatin interaction. J. Exp. Neurosci.
(2017) 11: 117.
Suzuki K, Sugihara G, Ouchi Y, Nakamura K,
Futatsubashi M, Takebayashi K, Yoshihara Y, Omata
K, Matsumoto K, Tsuchiya KJ, Iwata Y, Tsujii M,
Sugiyama T and Mori N. Microglial activation in
young adults with autism spectrum disorder. JAMA
Psychiatry (2013) 70: 49-58.
Reichel CM, Schwendt M, McGinty JF, Olive MF
and See RE. Neuropsychopharmacology (2011) 36:
782-92.
Halpin LE and Yamamoto BK. Peripheral ammonia
as a mediator of methamphetamine neurotoxicity. J.
Neurosci. (2012) 32: 13155–63.
Halpin LE, Northrop NA and Yamamoto BK.
Ammonia mediates methamphetamine-induced
increases in glutamate and excitotoxicity.
Neuropsychopharmacology (2014) 39: 1031-8.
Northrop NA, Halpin LE and Yamamoto BK. Peripheral
ammonia and blood brain barrier structure and function
after methamphetamine. Neuropharmacology (2016)
107: 18-26.
Virmani A, Gaetani F, Imam S, Binienda Z and
Ali S. Possible mechanism for the neuroprotective
effects of L-carnitine on methamphetamine-evoked
neurotoxicity. Ann. NY. Acad. Sci. (2003) 993: 197-
207.
Racette BA, Aschner M, Guilarte TR, Dydak U,
Criswell SR and Zheng W. Pathophysiology of
manganese-associated neurotoxicity. Neurotoxicology
(2012) 33: 881-6
Szepesi Z, Manouchehrian O, Bachiller S and Deierborg
T. Bidirectional microglia-neuron communication in
health and disease. Front. Cell Neurosci. (2018) 12:
323.
Cisneros IE and Ghorpade A. HIV-1, methamphetamine
and astrocyte glutamate regulation: combined
excitotoxic implications for neuro-AIDS. Curr. HIV
Res. (2012) 10: 392-406.
Kitamura O. Detection of methamphetamine
neurotoxicity in forensic autopsy cases. Leg. Med.
(Tokyo) (2009) 11 (Suppl 1): S63-5.
Farina M, Aschner M and Rocha JBT. Environmental
chemicals and neurotoxicity oxidative stress in MeHginduced neurotoxicity. Toxicol. Appl. Pharmacol.
(2011) 256: 405–17.
Han B, Zhang Y, Zhang Y, Bai Y, Chen X and
Huang R. Novel insight into circular RNA HECTD1
in astrocyte activation via autophagy by targeting
MIR142-TIPARP: implications for cerebral ischemic
stroke. Autophagy (2018) 14: 1164-84.
Niknazar S, Nahavandi A, Peyvandi AA, Peyvandi H,
Roozbahany NA and Abbaszadeh HA. Hippocampal
NR3C1 DNA methylation can mediate part of
preconception paternal stress effects in rat offspring.
Behav. Brain Res. (2017) 324: 71-6.
Enquan X, Jianuo L, Han L, Xiaobei W and Huangui
X. Role of microglia in methamphetamine-induced
neurotoxicity. Int. J. Physiol. Pathophysiol. Pharmacol.
(2017) 9: 84–100.
Arezoomandan R, Moradi M, Attarzadeh-Yazdi G,
Tomaz C and Haghparast A. Administration of activated
glial condition medium in the nucleus accumbens
extended extinction and intensified reinstatement
of Methamphetamine-induced conditioned place
preference. Brain Res. Bull. (2016) 125: 106-16.
Ho E and Pekny M. Glial fibrillary acidic protein
(GFAP) and the astrocyte intermediate filament system
in diseases of the central nervous system. Curr. Opin.
Cell Biol. (2015) 32: 121-30.
Kuczenski R, Everall IP, Crews L, Adame A, Grant
I and Masliah E. Escalating dose-multiple binge
methamphetamine exposure results in degeneration
of the neocortex and limbic system in the rat. Exp.
Neurol. (2007) 207: 42-51.