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Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System
Published on: May 12, 2011
Methotrexate induces astrocyte apoptosis by disrupting folate metabolism in the mouse juvenile central nervous system
Yanfei Shao1, Biqin Tan2, Jing Shi3
1Department of Pharmacy, Zhejiang Provincial People's Hospital, Hangzhou Medical College, Hangzhou 310014, China; Department of Pharmacy, Hangzhou Medical College, Hangzhou 310053, China.
Abstract:
Methotrexate (MTX) is a folic acid antagonist and widely used for acute lymphoblastic leukemia (ALL) in children. MTX is associated with acute and chronic neurotoxicity during treatment, however the underlying mechanism is still poorly understood. In this study we investigate whether MTX is neurovirulent to astrocytes in the Central Nervous System (CNS) of adolescent mice. We demonstrated that MTX induced severe cytotoxicity in C6 astrocyte-like cell line and rat primary cultures of astrocytes in a dose-dependent manner. Moreover, GFAP-labeled astrocyte cells significantly decreased in the mouse spinal cord and brain. Furthermore, protein levels of PARP and pro-Caspase-3 were reduced by MTX, indicating MTX-induced apoptosis leads to the astrocytes loss. Notably, overexpression of dihydrofolate reductase (DHFR) or exogenous addition of folate markedly reversed the astrocytes toxicity induced by MTX through activating folate metabolism pathway. Taken together, our study provides evidence for neurotoxic effect of MTX-induced astrocytes apoptosis both in vitro and in vivo with disruption of folate metabolism, and additional supplement of folate may provide novel approaches for alleviating the astrocytes toxicity induced by MTX in the clinic.
Insights
Methotrexate (MTX) causes neurotoxicity by inducing astrocyte apoptosis, impacting the central nervous system. Folate supplementation may mitigate this MTX-induced toxicity in patients.
Area of Science:
- Neuroscience
- Cell Biology
- Pharmacology
Background:
- Methotrexate (MTX) is a key treatment for childhood acute lymphoblastic leukemia (ALL).
- MTX is known to cause neurotoxicity, but the mechanisms remain unclear.
- Astrocytes play crucial roles in central nervous system (CNS) function and homeostasis.
Purpose of the Study:
- To investigate the neurovirulence of MTX on astrocytes in the CNS.
- To elucidate the cellular mechanisms underlying MTX-induced neurotoxicity.
Main Methods:
- In vitro studies using C6 astrocyte-like cell lines and primary rat astrocyte cultures.
- In vivo studies in adolescent mice, examining spinal cord and brain tissues.
- Assessment of astrocyte viability, GFAP expression, and apoptosis markers (PARP, pro-Caspase-3).
- Evaluation of the effects of dihydrofolate reductase (DHFR) overexpression and folate supplementation.
Main Results:
- MTX induced dose-dependent cytotoxicity in astrocytes in vitro.
- MTX significantly reduced GFAP-labeled astrocytes in the mouse CNS.
- MTX treatment led to apoptosis in astrocytes, evidenced by reduced PARP and pro-Caspase-3 levels.
- Overexpression of DHFR or addition of folate reversed MTX-induced astrocyte toxicity via folate metabolism.
Conclusions:
- MTX induces astrocyte apoptosis in vitro and in vivo, contributing to neurotoxicity.
- Disruption of folate metabolism is implicated in MTX-induced astrocyte damage.
- Folate supplementation presents a potential therapeutic strategy to alleviate MTX neurotoxicity.
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