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An ASK1-p38 signalling pathway mediates hydrogen peroxide-induced toxicity in NG108-15 neuronal cells
Koji Nomura1, Mercede Lee, Christina Banks
1Institute of Neuroscience and Psychology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
Abstract:
Reactive oxygen species (ROS) are believed to be involved in many forms of neurodegeneration, including ischaemic infarct damage and Alzheimer's disease. Despite the known involvement of p38 and JNK MAP kinases in mediating apoptosis and cell death in a variety of cell types, the details of the signalling pathways activated in neuronal cells by ROS are poorly characterised. Recently TAK1 (MAP3K7), a kinase upstream of JNK and p38, has attracted attention as a possible mediator of ischaemic cell death. This study tested the hypothesis that hydrogen peroxide (H2O2), which produces ROS, induces apoptosis in the NG108-15 neuronal cell line via activation of either TAK1 or the related kinase ASK1 (MAP3K5). H2O2 caused a concentration-dependent reduction in cell viability associated with caspase 3 activation. Loss of cell viability was inhibited by a selective caspase 3 inhibitor, and by the p38 inhibitor SB203580, but was not affected by the JNK inhibitor SP600125. The selective TAK1 inhibitor 5Z-7-oxozeaenol (5Z-7) exacerbated the loss of cell viability, whereas the ASK1 inhibitor NQDI-1 completely prevented caspase activation and cell death. These results show that pharmacological inhibition of ASK1 is neuroprotective, implicating an ASK1-p38 signalling pathway in ROS-induced apoptosis in neurones. The results also imply that the role of TAK1 may be neuroprotective rather than pro-degenerative.
Insights
Hydrogen peroxide (H2O2) triggers neuronal cell death via reactive oxygen species (ROS). Inhibiting ASK1 kinase protects against this apoptosis, suggesting a new therapeutic target for neurodegeneration.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Reactive oxygen species (ROS) are implicated in neurodegenerative diseases like Alzheimer's.
- Signaling pathways in neuronal ROS response, particularly involving MAP kinases, are not fully understood.
Purpose of the Study:
- To investigate if hydrogen peroxide (H2O2)-induced apoptosis in NG108-15 neuronal cells involves TAK1 or ASK1.
- To elucidate the specific roles of TAK1 and ASK1 in ROS-mediated neuronal cell death.
Main Methods:
- NG108-15 neuronal cells were treated with varying concentrations of H2O2.
- Cell viability was assessed, and caspase 3 activation was measured.
- Selective inhibitors for caspase 3, p38, JNK, TAK1, and ASK1 were used to determine pathway involvement.
Main Results:
- H2O2 induced a dose-dependent decrease in cell viability and activated caspase 3.
- ASK1 inhibition completely prevented caspase activation and cell death, indicating neuroprotection.
- TAK1 inhibition worsened cell death, suggesting a potentially neuroprotective role for TAK1.
Conclusions:
- ASK1 activation is a key mediator of ROS-induced apoptosis in neurons.
- An ASK1-p38 signaling pathway is implicated in H2O2-induced neuronal cell death.
- Targeting ASK1 offers a potential neuroprotective strategy against ROS-mediated damage.
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