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Morphine Induces Apoptosis, Inflammation, and Mitochondrial Oxidative Stress via Activation of TRPM2 Channel and Nitric Oxide Signaling Pathways in the Hippocampus

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Abstract

Morphine as an opioid is an important drug in the treatment of moderate to severe pain. Several stress factors via generation of nitric oxide (NO) and oxidative stress (OS) are responsible for the adverse effects of morphine-induced analgesia, addiction, and antinociceptive tolerance, including altered Ca2+ concentration, inflammation, OS, and release of apoptotic factors. TRPM2 is a Ca2+-permeable cation channel and it is activated by OS and NO. Hence, adverse effect of morphine addiction may occur via the OS and NO-induced TRPM2 activation. Because of the unclear etiology of morphine-induced adverse effects in the hippocampus, investigating the involvement of TRPM2 and NO synthetase (NOS) activations in the treatment of morphine-induced OS, apoptosis, and neuroinflammation is a major challenge. The hippocampal neuron of TRPM2 wild-type (TRPM2-WT) and knockout (TRPM2-KO) mice were divided into control, morphine, NOS inhibitor (L-NAME) + morphine, and TRPM2 channel blockers (ACA and 2-APB) + morphine. The morphine-induced increases of apoptosis, neuron death, OS, lipid peroxidation, caspase-3 and caspase-9, neuroinflammatory cytokines (IL-1β, TNF-α, IL-6), and Ca2+ levels in the hippocampal neuron of TRPM2-WT mouse were decreased by the L-NAME, ACA, and 2-APB treatments, although cell viability, neuron count, and reduced glutathione and glutathione peroxidase levels were increased by the treatments. However, the effects of morphine were not observed in the hippocampus of TRPM2-KO mice. Taken together, our data show that neurodegeneration adverse effects of morphine were induced by activation of TRPM2, and excessive generations of NO and OS. Thus, inhibition of TRPM2 may modulate morphine-induced neurodegeneration in the hippocampus.

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  • 22 June 2020

    The original version of this article unfortunately contained some mistake.

Abbreviations

[Ca2+]c :

Intracellular free calcium ion concentration

2-APB:

2-Aminoethoxydiphenyl borate

ACA:

N-(p-amylcinnamoyl) anthranilic acid

CPx:

Cumene hydroperoxide

L-NAME:

NG-nitro-l-arginine methyl ester

LSC:

Laser scan confocal

MDA:

Malondialdehyde

NAD+ :

Nicotinamide adenine dinucleotide

NO:

Nitric oxide

NOS:

Nitric oxide synthase

OS:

Oxidative stress

PARP-1 :

Poly [ADP-ribose] polymerase-1

rGSH:

Reduced glutathione

TRPM2 :

Transient receptor potential melastatin 2

TRPM2-KO:

TRPM2 knockout

TRPM2-WT:

TRPM2 wild type

VGCC:

Voltage-gated calcium channel

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Acknowledgments

The authors wish to thank technician Hulusi Gül (BSN Health, Analyses, Innovation, Consultancy, Organization, Agriculture and Industry Ltd., Göller Bölgesi Teknokenti, Isparta, Turkey) for helping with the animal experiments and LSC microscope. The cell culture, antioxidant, and cell viability analyses in the current study were performed in 3rd International Brain Research School, 25 June–1 July October 2018, Isparta, Turkey by Ö.O., M.K.Y., and Y.A. (http://2018.brs.org.tr/).

Funding

The study was supported by BSN Health, Analyses, Innovation, Consultancy, Organization, Agriculture and Industry Ltd., Isparta, Turkey (Project No. 2018-033). Coordinator of the project was M.N. There is no financial disclosure of the current study.

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Authors and Affiliations

  1. Department of Anesthesiology and Reanimation, Faculty of Medicine, Suleyman Demirel University, Isparta, Turkey

    Haci Ömer Osmanlıoğlu & Mustafa Kemal Yıldırım

  2. Department of Neurosurgery, Faculty of Medicine, Hatay Mustafa Kemal University, Hatay, Turkey

    Yener Akyuva

  3. Department of Biophysics, Faculty of Medicine, Suleyman Demirel University, Isparta, Turkey

    Kenan Yıldızhan & Mustafa Nazıroğlu

  4. Drug Discovery Unit, BSN Health, Analyses, Innovation, Consultancy, Organization, Agriculture and Industry Ltd, Göller Bölgesi Teknokenti, Isparta, Turkey

    Mustafa Nazıroğlu

  5. Neuroscience Research Center, Suleyman Demirel University, Isparta, Turkey

    Mustafa Nazıroğlu

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  1. Haci Ömer Osmanlıoğlu
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  2. Mustafa Kemal Yıldırım
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  3. Yener Akyuva
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Contributions

Ö.O. and M.N. formulated the hypothesis and M.N. was responsible for writing the report. K.Y. was responsible for isolating the hippocampus and analyzing the intracellular Ca2+ concentration. The cell culture, antioxidant, and cell viability analyses were performed by Ö.O. and M.K.Y. Y.A. also made critical revision of the manuscript.

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Correspondence to Mustafa Nazıroğlu.

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Ethical Approval

This article does not contain any studies with human participants performed by any of the authors. The study was approved by the Experimental Animal Research Center of Suleyman Demirel University (SDU), Isparta, Turkey (28 February 2019; Protocol No. 2019-03-01; applicant—Ö.O.).

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The original version of this article was revised: The first givenname of the first author has been added to read “Haci Ömer Osmanlıoğlu”.

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Osmanlıoğlu, H.Ö., Yıldırım, M.K., Akyuva, Y. et al. Morphine Induces Apoptosis, Inflammation, and Mitochondrial Oxidative Stress via Activation of TRPM2 Channel and Nitric Oxide Signaling Pathways in the Hippocampus. Mol Neurobiol 57, 3376–3389 (2020). https://doi.org/10.1007/s12035-020-01975-6

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