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Urolithin B protects PC12 cells against glutamate-induced toxicity

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Abstract

Background

The involvement of malfunctioning glutamate systems in various central nervous system (CNS) disorders is widely acknowledged. Urolithin B, known for its neuroprotective and antioxidant properties, has shown potential as a therapeutic agent for these disorders. However, little is known about its protective effects against glutamate-induced toxicity in PC12 cells. Therefore, in this study, for the first time we aimed to investigate the ability of Urolithin B to reduce the cytotoxic effects of glutamate on PC12 cells.

Methods

Different non-toxic concentrations of urolithin B were applied to PC12 cells for 24 h before exposure to glutamate (10 mM). The cells were then analyzed for cell viability, intracellular reactive oxygen species (ROS), cell cycle arrest, apoptosis, and the expression of Bax and Bcl-2 genes.

Results

The results of MTT assay showed that glutamate at a concentration of 10 mM and urolithin B at a concentration of 114 μM can reduce PC12 cell viability by 50%. However, urolithin B at non-toxic concentrations of 4 and 8 μM significantly reduced glutamate-induced cytotoxicity (p < 0.01). Interestingly, treatment with glutamate significantly enhanced the intracellular ROS levels and apoptosis rate in PC12 cells, while pre-treatment with non-toxic concentrations of urolithin B significantly reduced these cytotoxic effects. The results also showed that pre-treatment with urolithin B can decrease the Bax (p < 0.05) and increase the Bcl-2 (p < 0.01) gene expression, which was dysregulated by glutamate.

Conclusions

Taken together, urolithin B may play a protective role through reducing oxidative stress and apoptosis against glutamate-induced toxicity in PC12 cells, which merits further investigations.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Abbreviations

DMSO:

Dimethyl sulfoxide

qRT-PCR:

Quantitative real-time polymerase chain reaction

ROS:

Reactive oxygen species

CNS:

Central nervous system

MTT:

3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide

PI:

Propidium iodide

FBS:

Fetal bovine serum

TBHP:

Tert-butyl hydroperoxide

PD:

Parkinson’s disease

NO:

Nitric oxide

IL-6:

Interleukin-6

TNF-α:

Tumor necrosis factor-alpha

PGE2:

Prostaglandin E2

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Funding

This work’s authors recognize the following funding sources for their research, writing, and publication: Financial support for this project came from the Mashhad University of Medical Sciences (Grant No. 4011832).

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Authors and Affiliations

  1. Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Israa Aljabouri, Mehdi Rostami, Mahla Kazemian Kakhki, Abbas Alalikhan, Elaheh Gheybi, Ali Hakimi & Mohammad Soukhtanloo

  2. Cardiovascular Diseases Research Center, Birjand University of Medical Sciences, Birjand, Iran

    Farshad Mirzavi

  3. Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Iran

    Mohammad Soukhtanloo

Authors
  1. Israa Aljabouri
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  2. Mehdi Rostami
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  3. Farshad Mirzavi
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  4. Mahla Kazemian Kakhki
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  5. Abbas Alalikhan
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  7. Ali Hakimi
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  8. Mohammad Soukhtanloo
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Contributions

IA, MR, and AA contributed to the manuscript’s conception, design, acquisition, and drafting. FM, MKK, AH, and EG contributed to the interpretation and critically revised the manuscript. MS made distinctive contributions to the idea and demonstrated their specialized expertise to enhance the overall quality of the manuscript. All authors have thoroughly reviewed and provided their consent to the final version of the manuscript.

Corresponding author

Correspondence to Mohammad Soukhtanloo.

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There are no known possible conflicts of interest.

Ethics approval and informed consent

The ethics committee of Mashhad University of Medical Sciences, Mashhad, Iran, approved the ethical issues of this study. It was a cell-based study, with no human sample included.

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Aljabouri, I., Rostami, M., Mirzavi, F. et al. Urolithin B protects PC12 cells against glutamate-induced toxicity. Mol Biol Rep 51, 360 (2024). https://doi.org/10.1007/s11033-024-09236-8

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