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L-Ascorbate Protects Against Methamphetamine-Induced Neurotoxicity of Cortical Cells via Inhibiting Oxidative Stress, Autophagy, and Apoptosis

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Abstract

Methamphetamine (METH)-induced cell death contributes to the pathogenesis of neurotoxicity; however, the relative roles of oxidative stress, apoptosis, and autophagy remain unclear. L-Ascorbate, also called vitamin (Vit.) C, confers partial protection against METH neurotoxicity via induction of heme oxygenase-1. We further investigated the role of Vit. C in METH-induced oxidative stress, apoptosis, and autophagy in cortical cells. Exposure to lower concentrations (0.1, 0.5, 1 mM) of METH had insignificant effects on ROS production, whereas cells exposed to 5 mM METH exhibited ROS production in a time-dependent manner. We confirmed METH-induced apoptosis (by nuclear morphology revealed by Hoechst 33258 staining and Western blot showing the protein levels of pro-caspase 3 and cleaved caspase 3) and autophagy (by Western blot showing the protein levels of Belin-1 and conversion of microtubule-associated light chain (LC)3-I to LC3-II and autophagosome staining by monodansylcadaverine). The apoptosis as revealed by cleaved caspase-3 expression marked an increase at 18 h after METH exposure while both autophagic markers, Beclin 1 and LC3-II, marked an increase in cells exposed to METH for 6 and 24 h, respectively. Treating cells with Vit. C 30 min before METH exposure time-dependently attenuated the production of ROS. Vitamin C also attenuated METH-induced Beclin 1 and LC3-II expression and METH toxicity. Treatment of cells with Vit. C before METH exposure attenuated the expression of cleaved caspase-3 and reduced the number of METH-induced apoptotic cells. We suggest that the protective effect of Vit. C against METH toxicity might be through attenuation of ROS production, autophagy, and apoptosis.

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Acknowledgments

This work was supported in part by a grant (03C0720004A) to YNH from Taipei Medical University, Taiwan, and grants MOST-103-2321-B-038-002 and MOST-104-2320-B-038-057-MY3 from the Ministry of Sciences and Technology and MOHW104-TDU-B-212-124-001 from the Ministry of Health and Welfare surcharge of tobacco products, Taiwan, to JYW.

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Authors and Affiliations

  1. Department of Nursing, Hsin Sheng Junior College of Medical Care and Management, Taoyuan, Taiwan

    Ya-Ni Huang

  2. Graduate Institute of Medical Sciences and Department of Physiology, College of Medicine, Taipei Medical University, 250 Wu-Hsing Street, Taipei, 110, Taiwan

    Ling-Yu Yang, Jing-Ya Wang & Jia-Yi Wang

  3. Division of Orthopedics, Department of Surgery, Far Eastern Memorial Hospital, New Taipei City, Taiwan

    Chien-Cheng Lai

  4. Department of Neurosurgery, En Chu Kong Hospital, New Taipei City, Taiwan

    Chien-Tsai Chiu

  5. Comprehensive Cancer Center, Taipei Medical University, Taipei, Taiwan

    Jia-Yi Wang

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Huang, YN., Yang, LY., Wang, JY. et al. L-Ascorbate Protects Against Methamphetamine-Induced Neurotoxicity of Cortical Cells via Inhibiting Oxidative Stress, Autophagy, and Apoptosis. Mol Neurobiol 54, 125–136 (2017). https://doi.org/10.1007/s12035-015-9561-z

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