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The effects of mesenchymal stem cells transplantation on A1 neurotoxic reactive astrocyte and demyelination in the cuprizone model

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Abstract

Multiple sclerosis (MS), which is an autoimmune disease, is characterized by symptoms such as demyelination, axonal damage, and astrogliosis. As the most abundant type of glial cells, astrocytes play an important role in MS pathogenesis. Mesenchymal stem cells (MSCs) are a subset of stromal cells that have the potential for migration, immune-modulation, differentiation, remyelination, and neuroregeneration. Therefore, the present study evaluates the effects of MSC transplantation on A1 reactive astrocytes and the remyelination process in the cuprizone mouse model. The study used 30 male C57BL/6 mice, which were randomly distributed into three subgroups (n = 10), i.e., control, cuprizone, and transplanted MSCs groups. In order to generate a chronic demyelination model, the mice in the cuprizone group received food mixed with 0.2% cuprizone powder for 12 weeks. Then, 2 μl of DMEM containing approximately 3 × 105 DiI labeled cells was injected with a 4-min interval into the right lateral ventricle using a 10‐μl Hamilton syringe. After 2 weeks of cell transplantation, we used the rotarod test to evaluate the behavioral deficits, while the remyelination process was assessed by transmission electron microscopy (TEM) and Luxol Fast Blue (LFB) staining. We assessed the population of A1 astrocytes and oligodendrocytes using specific markers, such as C3, GFAP, and Olig2, using the immunefleurocent method. The pro-inflammatory and trophic factors were assessed by a real‐time polymerase chain reaction. According to our data, the specific marker of A1 astrocytes (C3) decreased in the MSCs group, while the number of oligodendrocytes significantly increased in this group compared to the cuprizone mice. Additionally, MSC was able to enhance the remyelination process after cuprizone usage, as shown by LFB and TEM images. The molecular results showed that MSCs could reduce pro-inflammatory factors, such as IL-1 and TNF-α, through the secretion of BDNF and TGF-β as trophic factors. The obtained results indicated that MSC could reduce demyelination and inflammation by decreasing A1 neurotoxic reactive astrocytes and neurotrophic and immunomodulatory factors secretion in the chronic cuprizone demyelination model.

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Fig. 1
Fig. 2

Adapted from Paxinos Franklin mouse brain Atlas. B Immunofluorescente image of DiI‐labeled MSCs (red) in CC 2 weeks after injection. C DAPI staining for the nuclei (blue) assessment. D The merged image of MSC with nuclei (arrow). DAPI: 4′,6‐diamidino‐2‐phenylindole; CC: corpus callosum; DiI: 1,1′‐dioctadecyl‐3,3,3′,3′‐ tetramethylindocarbocyanine perchlorate; MSCs: mesenchymal stem cells

Fig. 3
Fig. 4

Adapted from Paxinos Franklin mouse brain Atlas. B The LFB images exhibited that the demyelinated region (white color) increased in the brain of the CPZ group, while they decreased 2 weeks after MSCs injection. C Quantitative analysis of LFB images indicated that the number of myelinated fibers increased after MSC transplantation. Values are expressed as mean ± SEM. The significance is shown by **p ≤ .01 and ***p ≤ .001. LFB: luxol fast blue; CPZ: cuprizone; MSCs: mesenchymal stem cells; CC: corpus callosum; SEM: standard error of mean

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Acknowledgements

This study was supported by Tehran University of Medical Sciences and Health Services, Tehran, Iran.

Funding

This study was supported by a grant received from the Tehran University of Medical Sciences and Health Services, Tehran, Iran.

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Authors and Affiliations

  1. Department of Anatomy, Saveh University of Medical Sciences, Saveh, Iran

    Shirin Barati

  2. Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran

    Iraj Ragerdi Kashani

  3. Department of Anatomy, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Fatemeh Tahmasebi

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  1. Shirin Barati
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  2. Iraj Ragerdi Kashani
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  3. Fatemeh Tahmasebi
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Contributions

FT and IRK conceived the original idea and supervised the project and wrote the manuscript. SB carried out the experiment and wrote the manuscript and performed the analytic calculations. All authors discussed the results and contributed to the final manuscript.

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Correspondence to Fatemeh Tahmasebi.

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The authors had no conflict of interest regarding the publication of this paper.

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All experimental procedures were carried out in accordance with the guidelines of the Ethic Committee of Tehran University of Medical Science (TUMS), Tehran, Iran. All efforts were made to reduce the number of animals used and their suffering.

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Barati, S., Kashani, I.R. & Tahmasebi, F. The effects of mesenchymal stem cells transplantation on A1 neurotoxic reactive astrocyte and demyelination in the cuprizone model. J Mol Histol 53, 333–346 (2022). https://doi.org/10.1007/s10735-021-10046-6

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  • DOI: https://doi.org/10.1007/s10735-021-10046-6

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