Abstract
Mitoxantrone (MTX) is a topoisomerase II inhibitor used to treat a wide range of tumors and multiple sclerosis but associated with potential neurotoxic effects mediated by hitherto poorly understood mechanisms. In adult male CD-1 mice, the underlying neurotoxic pathways of a clinically relevant cumulative dose of 6 mg/kg MTX was evaluated after biweekly administration for 3 weeks and sacrifice 1 week after the last administration was undertaken. Oxidative stress, neuronal damage, apoptosis, and autophagy were analyzed in whole brain, while coronal brain sections were used for a closer look in the hippocampal formation (HF) and the prefrontal cortex (PFC), as these areas have been signaled out as the most affected in ‘chemobrain’. In the whole brain, MTX-induced redox imbalance shown as increased endothelial nitric oxide synthase and reduced manganese superoxide dismutase expression, as well as a tendency to a decrease in glutathione levels. MTX also caused diminished ATP synthase β expression, increased autophagic protein LC3 II and tended to decrease p62 expression. Postsynaptic density protein 95 expression decreased in the whole brain, while hyperphosphorylation of Tau was seen in PFC. A reduction in volume was observed in the dentate gyrus (DG) and CA1 region of the HF, while GFAP-ir astrocytes increased in all regions of the HF except in the DG. Apoptotic marker Bax increased in the PFC and in the CA3 region, whereas p53 decreased in all brain areas evaluated. MTX causes damage in the brain of adult CD-1 mice in a clinically relevant cumulative dose in areas involved in memory and cognition.
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The data presented in this study are available on request from the corresponding authors.
Abbreviations
- ATG5:
-
Autophagy-related 5 protein
- ATP:
-
Adenosine triphosphate
- BBB:
-
Blood–brain barrier
- CNS:
-
Central nervous system
- DG:
-
Dentate gyrus
- eNOS:
-
Endothelial nitric oxide synthase
- GFAP:
-
Glial fibrillary acidic protein
- GSH:
-
Reduced glutathione
- GSK-3β:
-
Glycogen synthase kinase 3 beta
- GSSG:
-
Oxidized glutathione
- HF:
-
Hippocampal formation
- HSP27:
-
Heat shock protein 27
- iNOS:
-
Inducible nitric oxide synthase
- ir:
-
Immunoreactive
- IV:
-
Intravenously
- LC3:
-
Microtubule-associated protein 1 light chain 3
- MnSOD:
-
Manganese superoxide dismutase
- MTX:
-
Mitoxantrone
- NF-κB:
-
Nuclear factor kappa B
- OD:
-
Optical density
- PFC:
-
Prefrontal cortex
- PSD95:
-
Postsynaptic density protein 95
- pTau:
-
Phosphorylated Tau
- SD:
-
Standard deviation
- tGSH:
-
Total glutathione
- TNF-α:
-
Tumor necrosis alpha
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Funding
This work is financed by national funds from Fundação para a Ciência e a Tecnologia (FCT), I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences (UCIBIO) and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB and through the project EXPL/MEDFAR/0203/2021. A. Dias-Carvalho acknowledges FCT and UCIBIO for her PhD grant (UI/BD/151318/2021). V.M.C acknowledges FCT for her grant (SFRH/BPD/110001/2015) that was funded by national funds through FCT under the Norma Transitória–DL57/2016/CP1334/CT0006. A.R.-M. acknowledges FCT for her grant SFRH/BD/129359/2017.
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ADC and VMC conceived and designed the experiments; ADC and MF conducted the experiments and data analysis; ARM performed the in vivo experiments; SIS performed the stereological and morphometric analysis; ADC and MF drafted the article; ARM, RF, MLB, ED, SIS, JPC, FC, and VMC critically reviewed the article. All authors have read and agreed to the published version of the manuscript.
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The study was conducted according to the guidelines of the Declaration of Helsinki and approved by the Ethics Committee of the local animal welfare body (ICBAS-UP ORBEA) and the Portuguese national authority for animal health (DGAV, process no. 0421/000/000/2016).
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Dias-Carvalho, A., Ferreira, M., Reis-Mendes, A. et al. Chemobrain: mitoxantrone-induced oxidative stress, apoptotic and autophagic neuronal death in adult CD-1 mice. Arch Toxicol 96, 1767–1782 (2022). https://doi.org/10.1007/s00204-022-03261-x
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DOI: https://doi.org/10.1007/s00204-022-03261-x