L-Ascorbate attenuates methamphetamine neurotoxicity through enhancing the induction of endogenous heme oxygenase-1

Ya Ni Huang, Jiz Yuh Wang, Ching Tien Lee, Chih Hung Lin, Chien Cheng Lai, Jia Yi Wang

Research output: Contribution to journalArticle

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Abstract

Methamphetamine (METH) is a drug of abuse which causes neurotoxicity and increased risk of developing neurodegenerative diseases. We previously found that METH induces heme oxygenase (HO)-1 expression in neurons and glial cells, and this offers partial protection against METH toxicity. In this study, we investigated the effects of l-ascorbate (vitamin C, Vit. C) on METH toxicity and HO-1 expression in neuronal/glial cocultures. Cell viability and damage were evaluated by 3-(4,5-dimethylthianol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) release, respectively. Neuronal and glial localization of HO-1 were identified by double immunofluorescence staining. Reactive oxygen species (ROS) production was measured using the fluorochrome 2',7'-dichlorofluorescin diacetate. HO-1 mRNA and protein expression were examined by RT-qPCR and Western blotting, respectively. Results show that Vit. C induced HO-1 mRNA and protein expressions in time- and concentration-dependent manners. Inhibition of p38 mitogen-activated protein kinase (MAPK) but not extracellular signal-regulated kinase (ERK) significantly blocked induction of HO-1 by Vit. C. HO-1 mRNA and protein expressions were significantly elevated by a combination of Vit. C and METH, compared to either Vit. C or METH alone. Pretreatment with Vit. C enhanced METH-induced HO-1 expression and attenuated METH-induced ROS production and neurotoxicity. Pharmacological inhibition of HO activity abolished suppressive effects of Vit. C on METH-induced ROS production and attenuated neurotoxicity. We conclude that induction of HO-1 expression contributes to the attenuation of METH-induced ROS production and neurotoxicity by Vit. C. We suggest that HO-1 induction by Vit. C may serve as a strategy to alleviate METH neurotoxicity.

Original languageEnglish
Pages (from-to)241-252
Number of pages12
JournalToxicology and Applied Pharmacology
Volume265
Issue number2
DOIs
Publication statusPublished - Dec 1 2012

Fingerprint

Heme Oxygenase-1
Methamphetamine
Ascorbic Acid
Reactive Oxygen Species
Neuroglia
Messenger RNA
Toxicity
Neurodegenerative diseases
Heme Oxygenase (Decyclizing)
Proteins
Extracellular Signal-Regulated MAP Kinases
Street Drugs
p38 Mitogen-Activated Protein Kinases
Coculture Techniques
Bromides
Fluorescent Dyes
L-Lactate Dehydrogenase
Neurodegenerative Diseases
Neurons
Fluorescent Antibody Technique

Keywords

  • Heme oxygenase-1
  • Methamphetamine
  • P38 MAPK
  • ROS
  • Vitamin C

ASJC Scopus subject areas

  • Pharmacology
  • Toxicology

Cite this

L-Ascorbate attenuates methamphetamine neurotoxicity through enhancing the induction of endogenous heme oxygenase-1. / Huang, Ya Ni; Wang, Jiz Yuh; Lee, Ching Tien; Lin, Chih Hung; Lai, Chien Cheng; Wang, Jia Yi.

In: Toxicology and Applied Pharmacology, Vol. 265, No. 2, 01.12.2012, p. 241-252.

Research output: Contribution to journalArticle

Huang, Ya Ni ; Wang, Jiz Yuh ; Lee, Ching Tien ; Lin, Chih Hung ; Lai, Chien Cheng ; Wang, Jia Yi. / L-Ascorbate attenuates methamphetamine neurotoxicity through enhancing the induction of endogenous heme oxygenase-1. In: Toxicology and Applied Pharmacology. 2012 ; Vol. 265, No. 2. pp. 241-252.
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AB - Methamphetamine (METH) is a drug of abuse which causes neurotoxicity and increased risk of developing neurodegenerative diseases. We previously found that METH induces heme oxygenase (HO)-1 expression in neurons and glial cells, and this offers partial protection against METH toxicity. In this study, we investigated the effects of l-ascorbate (vitamin C, Vit. C) on METH toxicity and HO-1 expression in neuronal/glial cocultures. Cell viability and damage were evaluated by 3-(4,5-dimethylthianol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) release, respectively. Neuronal and glial localization of HO-1 were identified by double immunofluorescence staining. Reactive oxygen species (ROS) production was measured using the fluorochrome 2',7'-dichlorofluorescin diacetate. HO-1 mRNA and protein expression were examined by RT-qPCR and Western blotting, respectively. Results show that Vit. C induced HO-1 mRNA and protein expressions in time- and concentration-dependent manners. Inhibition of p38 mitogen-activated protein kinase (MAPK) but not extracellular signal-regulated kinase (ERK) significantly blocked induction of HO-1 by Vit. C. HO-1 mRNA and protein expressions were significantly elevated by a combination of Vit. C and METH, compared to either Vit. C or METH alone. Pretreatment with Vit. C enhanced METH-induced HO-1 expression and attenuated METH-induced ROS production and neurotoxicity. Pharmacological inhibition of HO activity abolished suppressive effects of Vit. C on METH-induced ROS production and attenuated neurotoxicity. We conclude that induction of HO-1 expression contributes to the attenuation of METH-induced ROS production and neurotoxicity by Vit. C. We suggest that HO-1 induction by Vit. C may serve as a strategy to alleviate METH neurotoxicity.

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