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Role of apoptosis and autophagy in mediating tramadol-induced neurodegeneration in the rat hippocampus

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Abstract

Background

Tramadol (TRA) is an analgesic prescribed for treating mild to moderate pains, the abuse of which has increased in recent years. Chronic tramadol consumption produces neurotoxicity, although the mechanisms are unclear. The present study investigated the involvement of apoptosis and autophagy signaling pathways and the mitochondrial system in TRA-induced neurotoxicity.

Materials and methods

Sixty adult male Wistar rats were divided into five groups that received standard saline or TRA in doses of 25, 50, 75, 100, or 150 mg/kg intraperitoneally for 21 days. On the 22nd day, the Open Field Test (OFT) was conducted. Jun N-Terminal Kinase (JNK), B-cell lymphoma-2 (Bcl-2), Beclin1, and Bcl-2-like protein 4 (Bax) proteins and tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) were measured in rat hippocampal tissue.

Results

TRA at doses 75, 100, and 150 mg/kg caused locomotor dysfunction in rats and increased total and phosphorylated forms of JNK and Beclin-1, Bax, and Caspase-3. TRA at the three higher doses also increased the phosphorylated (inactive) form of Bcl-2 level while decreasing the unphosphorylated (active) form of Bcl-2. Similarly, the protein levels of TNF-α and IL-1β were increased dose-dependently. The mitochondrial respiratory chain enzymes were reduced at the three higher doses of TRA.

Conclusion

TRA activated apoptosis and autophagy via modulation of TNF-α or IL-1β/JNK/Bcl-2/Beclin1 and Bcl-2/Bax signaling pathways and dysfunction of mitochondrial respiratory chain enzymes.

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Data Availability

The datasets are available from the corresponding author upon reasonable request.

Abbreviations

Akt :

Protein kinase B (PKB)

ANOVA :

Analysis of variances

Bax :

Bcl-2-like protein 4

Bcl-2 :

B-cell lymphoma-2

BSA :

Bovine serum albumin

EGTA :

ethylene glycol tetra-acetic acid

EPM :

Elevated Plus Maze

GSSG :

Glutathione in oxidized form

ELISA :

enzyme-linked immunosorbent assay

EPM :

Elevated Plus Maze

FAAD :

Fas Associated Via Death Domain

FAS :

Fas cell surface death receptor

FST :

Forced Swim Test

GSH :

reducing type of glutathione

HPLC :

High-performance liquid chromatography

H 2 SO 4 :

Sulphoric Acid

IHC :

Immunohistochemistry

IL-1β :

interleukin 1 beta

iNOS :

inducible nitric oxide synthase

JNKs :

c-Jun N-terminal kinases

MAPK :

Mitogen-activated protein kinase

MDA :

Malondialdehyde

MgCl2 :

magnesium chloride

MPO :

Myeloperoxidase

mTOR :

Mammalian target of rapamycin

NaCl :

Sodium Chloride

NADPH :

nicotinamide adenine dinucleotide phosphate

NaH2PO4 :

Sodium dihydrogen phosphate

NF-κB :

nuclear factor kappa B

NO :

Nitric Oxide

NOAEL :

No Observed Adverse Effect Level

OFT :

Open Field Test

PC :

personal computer

ROS :

Reactive Oxygen Species

SEM :

Standard error of the mean

TMB substrate :

3,3’,5,5’-Tetramethyl –benzidine

TNF-α :

tumor necrosis factor alpha

TRAIL/Apo2L :

Tumor necrosis factor-related apoptosis-inducing ligand

TRA :

Tramadol

TST :

Tail Suspension Test

UVRAG :

UV radiation resistance-related gene

WB :

Western blotting

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Funding

The current work was supported by the Chronic Respiratory Disease Research Center (CRDRC), National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran.

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Authors and Affiliations

  1. School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Mina Gholami

  2. University of South Florida College of Public Health, Tampa, FL, USA

    A. Wallace Hayes

  3. Institute for Integrative Toxicology, Michigan State University, East Lansing, MI, USA

    A. Wallace Hayes

  4. Chronic Respiratory Disease Research Center (CRDRC), National Research Institute of Tuberculosis and Lung Diseases (NRITLD), Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Hamidreza Jamaati & Majid Motaghinejad

  5. Research Group in Community Nutrition and Oxidative Stress (NUCOX), University of Balearic Islands and, Health Research Institute of Balearic Islands (IdISBa), Palma de Mallorca, Spain

    Antoni Sureda

  6. CIBER Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III (ISCIII), Madrid, Spain

    Antoni Sureda

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  1. Mina Gholami
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  2. A. Wallace Hayes
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Correspondence to Majid Motaghinejad.

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None declared.

Ethical approval

All experimental procedure of this research project was approved by the research committee in Masih Daneshvari Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran. (Research Protocol and ethical code number = IR.SBMU.NRITLD.REC.1401.105).

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Gholami, M., Hayes, A.W., Jamaati, H. et al. Role of apoptosis and autophagy in mediating tramadol-induced neurodegeneration in the rat hippocampus. Mol Biol Rep 50, 7393–7404 (2023). https://doi.org/10.1007/s11033-023-08641-9

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