Elsevier

Free Radical Biology and Medicine

Volume 131, 1 February 2019, Pages 408-431
Free Radical Biology and Medicine

Original article
Astrocytic mobilization of glutathione peroxidase-1 contributes to the protective potential against cocaine kindling behaviors in mice via activation of JAK2/STAT3 signaling

https://doi.org/10.1016/j.freeradbiomed.2018.12.027Get rights and content

Highlights

  • GPx-1 overexpression protects against oxidative burdens induced by cocaine.

  • Anti-convulsive potentials of GPx-1 require JAK2/STAT3/GFAP signaling process.

  • The activation of GFAP requires GPx-1 overexpression gene.

  • AG490 or astrocytic inhibition increased Iba-1-labeld microglial activity.

  • AG490 or astrocytic inhibition counteracted GPx-1-mediated protective potentials.

Abstract

Compelling evidence indicates that oxidative stress contributes to cocaine neurotoxicity. The present study was performed to elucidate the role of the glutathione peroxidase-1 (GPx-1) in cocaine-induced kindling (convulsive) behaviors in mice. Cocaine-induced convulsive behaviors significantly increased GPx-1, p-IkB, and p-JAK2/STAT3 expression, and oxidative burdens in the hippocampus of mice. There was no significant difference in cocaine-induced p-IkB expression between non-transgenic (non-TG) and GPx-1 overexpressing transgenic (GPx-1 TG) mice, but significant differences were observed in cocaine-induced p-JAK2/STAT3 expression and oxidative stress between non-TG and GPx-1 TG mice. Cocaine-induced glial fibrillary acidic protein (GFAP)-labeled astrocytic level was significantly higher in the hippocampus of GPx-1 TG mice. Triple-labeling immunocytochemistry indicated that GPx-1-, p-STAT3-, and GFAP-immunoreactivities were co-localized in the same cells. AG490, a JAK2/STAT3 inhibitor, but not pyrrolidone dithiocarbamate, an NFκB inhibitor, significantly counteracted GPx-1-mediated protective potentials (i.e., anticonvulsant-, antioxidant-, antiapoptotic-effects). Genetic overexpression of GPx-1 significantly attenuated proliferation of Iba-1-labeled microglia induced by cocaine in mice. However, AG490 or astrocytic inhibition (by GFAP antisense oligonucleotide and α-aminoadipate) significantly increased Iba-1-labeled microglial activity and M1 phenotype microglial mRNA levels, reflecting that proinflammatory potentials were mediated by AG490 or astrocytic inhibition. This microglial activation was less pronounced in GPx-1 TG than in non-TG mice. Furthermore, either AG490 or astrocytic inhibition significantly counteracted GPx-1-mediated protective potentials. Therefore, our results suggest that astrocytic modulation between GPx-1 and JAK2/STAT3 might be one of the underlying mechanisms for protecting against convulsive neurotoxicity induced by cocaine.

Introduction

Cocaine has been shown to possess convulsant effects in both rodents [1], [2] and human [3], [4]. The doses required to induce convulsant activity in rodents are quite high [5], [6], [7], and are comparable to doses of cocaine clinically associated with a drug overdose [8]. It is probable that a significant number of the clinical reports of cocaine-induced seizures are the result of the acute administration of high doses of cocaine [9], [10]. Studies indicated that 30% of chronic cocaine users exhibit memory impairments [10], and 7.8% of them experience cocaine-associated seizures [11]. Another retrospective survey documented that 65% of emergency room admissions for illicit drug-induced seizures were associated with cocaine abuse [12]. Furthermore, the repeated and intermittent administration of cocaine potentiated susceptibility to cocaine-induced seizures [5], [13]. This sensitization to the convulsant effects of cocaine has been termed “cocaine kindling” [5], [14] because of its similarities to other pharmacological and electric kindling processes. Previous report showed that reactive oxygen species (ROS) were produced after a single and repetitive administration of cocaine at low doses, indicating that oxidative burdens are involved in cocaine-mediated pharmacological action [15].

Glutathione peroxidase (GPx) is the most important component in the glutathione-mediated antioxidant system. GPx catalyzes the degradation of hydrogen peroxide (H2O2) and other hydroperoxides into water and alcohols, respectively [16]. Since catalase is mainly expressed in the peroxisomes and its activity is very low in the brain [17], GPx is regarded as the major antioxidant enzyme in the brain [18]. A selenium-dependent GPx-1 is the most abundant all of GPx isoforms (from GPx-1 to GPx-8) that have been identified [19].

GSH is a physiological substrate for GPx-1 in coping with ROS [20]. The GSH system is central to lipid peroxide removal [21]. We demonstrated that induction of GSH system is essential for preventing convulsive behaviors [22]. Interestingly, compensative induction of nuclear factor E2-related factor-2 (Nrf-2) gene induced by cocaine was correlated with antioxidant activity, while inhibition of Nrf-2 caused cell death through decreased GSH [23]. Therefore, it is plausible that impaired Nrf-1/GPx-1/GSH-system is important for inducing oxidative burdens elicited by cocaine.

Nuclear factor kappa B (NFκB) is a sensor of oxidative stress. ROS are critical intermediates for various NFκB-activating signals; when activated, NFκB promotes expression of proteins that participate as central regulators of immune, inflammatory, and apoptotic processes [24]. Previous reports indicate that NF-kB signaling might be involved in cocaine-induced memory impairment [25] and behavioral plasticity [26], suggesting that cocaine treatment induces oxidative burdens in the brain.

It is well-known that the signal transducers and activators of transcription 3 (STAT3) plays an important role in regulating gene expression, specifically increasing the expression levels of genes may be critical for cell survival (e.g., Bcl-2, and Bcl-xL), cell proliferation (e.g., c-Myc), cell-cycle progression (e.g., cyclin D1), angiogenesis (e.g., VEGF), and epileptogenesis [27]. Activation of the Janus kinase (JAK)/STAT pathway and specific phosphorylation of STAT3 were observed after the seizures induced by kainate [28].

Evidence has showed that repeated cocaine administration leads to reactive astrocytosis, as labeled by glial fibrillary acidic protein (GFAP) [29], suggesting that astrocytes are involved in the compensative mechanism against cocaine abuse. In addition, Little et al. [30] demonstrated that the number of activated microglia is significantly higher among cocaine users.

Therefore, the present study was designed to elucidate the role of GPx-1 gene in the cocaine-induced kindling (convulsive) behaviors, to investigate whether GPx-1 gene affects NFκB or JAK2/STAT3 signaling, and to investigate glial modulation in the cocaine-induced convulsive behaviors. We propose here that interactive modulation between JAK2/STAT3 signaling and astrocytic modulation of GPx-1 is critical for anticonvulsive/protective potential against cocaine insult.

Section snippets

Animal

All animals were treated in accordance with the National Institutes of Health (NIH) Public Health Service Policy on Humane Care and Use of Laboratory Animals (2015 Edition; grants.nih.gov/grants/olaw/references/PHSPolicyLabAnimals.pdf) and in accordance with the Institute for Laboratory Animal Research (ILAR) Guidelines for the Care and Use of Laboratory Animals (8th Edition; grants.nih.gov/grants/olaw/Guide-for-the-care-and-use-of-laboratory-animals.pdf). Glutathione peroxidase-1

Repeated treatment with cocaine resulted in significant increases in GPx-1 expression

As shown in Fig. 1A of the experimental design, we examined GPx-1 expression 1 h, 6 h, 12 h, 1 d, 3 d, 7 d, and 14 d after the final cocaine administration in the hippocampus of non-TG and GPx-1 TG mice. Treatment with cocaine resulted in an initial increase in GPx-1 expression 1 h post-cocaine (p < 0.05 vs. respective saline). Cocaine-induced increase in GPx-1 expression is the most prominent 1d post-cocaine in non-TG (p < 0.01 vs. respective saline control) and GPx-1 TG (p < 0.01 vs.

Discussion

The investigation of cocaine-kindling behaviors has been suggested for studying the toxicity associated with cocaine abuse. Cocaine-induced kindling profile in this study is comparable to that described by Miller et al. [5]. We propose here that cocaine-induced compensative induction of GPx-1 may be an important defensive mechanism, and that genetic overexpression of GPx-1 protects against cocaine-induced neurotoxic damage, suggesting that GPx-1 gene affords potential protection against cocaine

Acknowledgements

This study was supported by a grant (14182MFDS979) from the Korea Food and Drug Administration, Republic of Korea, and by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT, Republic of Korea (#NRF-2017R1A2B1003346 and #NRF-2016R1A1A1A05005201). Huynh Nhu Mai and Naveen Sharma were supported by the BK21 PLUS program, National Research Foundation of Korea, Republic of Korea. The English in this manuscript has been

Conflict of interest

The authors declare no conflict of interest.

References (96)

  • H.L. Grabenstatter et al.

    The effect of STAT3 inhibition on status epilepticus and subsequent spontaneous seizures in the pilocarpine model of acquired epilepsy

    Neurobiol. Dis.

    (2014)
  • J.S. Choi et al.

    Upregulation of gp130 and differential activation of STAT and p42/44 MAPK in the rat hippocampus following kainic acid-induced seizures

    Brain Res. Mol. Brain Res.

    (2003)
  • K.Y. Little et al.

    Decreased brain dopamine cell numbers in human cocaine users

    Psychiatry Res.

    (2009)
  • W.H. Cheng et al.

    Overexpression of cellular glutathione peroxidase does not affect expression of plasma glutathione peroxidase or phospholipid hydroperoxide glutathione peroxidase in mice offered diets adequate or deficient in selenium

    J. Nutr.

    (1997)
  • E.J. Shin et al.

    Genetic overexpression of glutathione peroxidase-1 attenuates microcystin-leucine-arginine-induced memory impairment in mice

    Neurochem. Int.

    (2018)
  • B.K. Kim et al.

    IL-6 attenuates trimethyltin-induced cognitive dysfunction via activation of JAK2/STAT3, M1 mAChR and ERK signaling network

    Cell Signal.

    (2013)
  • T.H. Tran et al.

    Repeated exposure to far infrared ray attenuates acute restraint stress in mice via inhibition of JAK2/STAT3 signaling pathway by induction of glutathione peroxidase-1

    Neurochem. Int.

    (2016)
  • D.K. Dang et al.

    PKCdelta-dependent p47phox activation mediates methamphetamine-induced dopaminergic neurotoxicity

    Free Radic. Biol. Med.

    (2018)
  • H.N. Mai et al.

    Exposure to far-infrared rays attenuates methamphetamine-induced recognition memory impairment via modulation of the muscarinic M1 receptor, Nrf2, and PKC

    Neurochem. Int.

    (2018)
  • P.T. Nguyen et al.

    Role of dopamine D1 receptor in 3-fluoromethamphetamine-induced neurotoxicity in mice

    Neurochem. Int.

    (2018)
  • H.Y. Tran et al.

    Protective potential of IL-6 against trimethyltin-induced neurotoxicity in vivo

    Free Radic. Biol. Med.

    (2012)
  • D.K. Dang et al.

    Genetic or pharmacological depletion of cannabinoid CB1 receptor protects against dopaminergic neurotoxicity induced by methamphetamine in mice

    Free Radic. Biol. Med.

    (2017)
  • J.A. Miller et al.

    Repeated exposure to low doses of kainic acid activates nuclear factor kappa B (NF-kappaB) prior to seizure in transgenic NF-kappaB/EGFP reporter mice

    Neurotoxicology

    (2014)
  • M.E. Hamby et al.

    Reactive astrocytes as therapeutic targets for CNS disorders

    Neurotherapeutics

    (2010)
  • F. Besnard et al.

    Multiple interacting sites regulate astrocyte-specific transcription of the human gene for glial fibrillary acidic protein

    J. Biol. Chem.

    (1991)
  • L.M. Shaffer et al.

    Amyloid beta protein (A beta) removal by neuroglial cells in culture

    Neurobiol. Aging

    (1995)
  • T. Wyss-Coray et al.

    Inflammation in neurodegenerative disease-a double-edged sword

    Neuron

    (2002)
  • R.M. Post

    Do the epilepsies, pain syndromes, and affective disorders share common kindling-like mechanisms?

    Epilepsy Res.

    (2002)
  • J. Wetherington et al.

    Astrocytes in the epileptic brain

    Neuron

    (2008)
  • O. Devinsky et al.

    Glia and epilepsy: excitability and inflammation

    Trends Neurosci.

    (2013)
  • A. Varin et al.

    Alternative activation of macrophages: immune function and cellular biology

    Immunobiology

    (2009)
  • E. Castilla-Ortega et al.

    The impact of cocaine on adult hippocampal neurogenesis: potential neurobiological mechanisms and contributions to maladaptive cognition in cocaine addiction disorder

    Biochem. Pharmacol.

    (2017)
  • A. Mansour et al.

    A comparison of D1 receptor binding and mRNA in rat brain using receptor autoradiographic and in situ hybridization techniques

    Neuroscience

    (1992)
  • Y.J. Lee et al.

    Increased expression of glial cell line-derived neurotrophic factor (GDNF) in the brains of scrapie-infected mice

    Neurosci. Lett.

    (2006)
  • C.J. Messer et al.

    Role for GDNF in biochemical and behavioral adaptations to drugs of abuse

    Neuron

    (2000)
  • M. Niwa et al.

    An inducer for glial cell line-derived neurotrophic factor and tumor necrosis factor-alpha protects against methamphetamine-induced rewarding effects and sensitization

    Biol. Psychiatry

    (2007)
  • C.C. Chao et al.

    Neuroprotective mechanism of glial cell line-derived neurotrophic factor on dopamine neurons: role of antioxidation

    Neuropharmacology

    (1999)
  • F. Cao et al.

    Conditional deletion of Stat3 promotes neurogenesis and inhibits astrogliogenesis in neural stem cells

    Biochem. Biophys. Res. Commun.

    (2010)
  • F.W. Wang et al.

    Roles of activated astrocyte in neural stem cell proliferation and differentiation

    Stem Cell Res.

    (2011)
  • X. Chen et al.

    Long-acting cocaine hydrolase for addiction therapy

    Proc. Natl. Acad. Sci. USA

    (2016)
  • R.H. Purcell et al.

    Effects of an epilepsy-causing mutation in the SCN1A sodium channel gene on cocaine-induced seizure susceptibility in mice

    Psychopharmacolog

    (2013)
  • T.R. Chang et al.

    Cocaine use as an independent predictor of seizures after aneurysmal subarachnoid hemorrhage

    J. Neurosurg.

    (2016)
  • K.A. Miller et al.

    Pharmacological and behavioral characterization of cocaine-kindled seizures in mice

    Psychopharmacology

    (2000)
  • C.U. Nnadi et al.

    Neuropsychiatric effects of cocaine use disorders

    J. Natl. Med. Assoc.

    (2005)
  • M. Vonmoos et al.

    Cognitive dysfunctions in recreational and dependent cocaine users: role of attention-deficit hyperactivity disorder, craving and early age at onset

    Br. J. Psychiatry

    (2013)
  • N. Majlesi et al.

    Cocaine-associated seizures and incidence of status epilepticus

    West. J. Emerg. Med.

    (2010)
  • D.H. Moak et al.

    Alcohol-related seizures and the kindling effect of repeated detoxifications: the influence of cocaine

    Alcohol Alcoholism

    (1996)
  • R.M. Kaminski et al.

    Effects of cocaine-kindling on the expression of NMDA receptors and glutamate levels in mouse brain

    Neurochem. Res.

    (2011)
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