Abstract
Inflammatory conversion of murine astrocytes correlates with the activation of various MAPK, and inhibition of terminal MAPKs like JNK or p38 dampens the inflammatory reaction. Mixed lineage kinases (MLKs), a family of MAPK kinase kinases, may therefore be involved in astrocyte inflammation. In this study, we explored the effect of the MLK inhibitors CEP-1347 and CEP-11004 on the activation of murine astrocytes by either TNF plus IL-1 or by a complete cytokine mix containing additional IFN-gamma. The compounds blocked NO-, PG-, and IL-6 release with a median inhibitory concentration of approximately 100 nM. This activity correlated with a block of the JNK and the p38 pathways activated in complete cytokine mix-treated astrocytes. Although CEP-1347 did not affect the activation of NF-kappaB, it blocked the expression of cyclooxygenase-2 and inducible NO synthase at the transcriptional level. Quantitative transcript profiling of 17 inflammation-linked genes revealed a specific modulation pattern of astrocyte activation by MLK inhibition, for instance, characterized by up-regulation of the anti-stress factors inhibitor of apoptosis protein-2 and activated transcription factor 4, no effect on manganese superoxide dismutase and caspase-11, and down-regulation of major inflammatory players like TNF, GM-CSF, urokinase-type plasminogen activator, and IL-6. In conclusion, MLK inhibitors like CEP-1347 are highly potent astrocyte immune modulators with a novel spectrum of activity.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Astrocytes / drug effects*
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Astrocytes / enzymology
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Astrocytes / immunology
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Blotting, Western
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Carbazoles / pharmacology*
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Cells, Cultured
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Cyclooxygenase 2
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Dose-Response Relationship, Drug
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Enzyme Activation / drug effects
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Enzyme Activation / immunology
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Enzyme Inhibitors / pharmacology*
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Gene Expression / drug effects
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Immunoassay
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In Situ Hybridization
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Indoles / pharmacology*
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Inflammation / immunology*
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Interleukin-6 / metabolism
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Isoenzymes / drug effects
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Isoenzymes / metabolism
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JNK Mitogen-Activated Protein Kinases*
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MAP Kinase Kinase 4
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MAP Kinase Kinase Kinases / drug effects*
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MAP Kinase Kinase Kinases / immunology
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Mice
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Mitogen-Activated Protein Kinase Kinases / drug effects
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Mitogen-Activated Protein Kinase Kinases / metabolism
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NF-kappa B / drug effects
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NF-kappa B / metabolism
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Nitric Oxide / metabolism
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Nitric Oxide Synthase / drug effects
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Nitric Oxide Synthase / metabolism
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Nitric Oxide Synthase Type II
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Polymerase Chain Reaction
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Prostaglandin-Endoperoxide Synthases / drug effects
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Prostaglandin-Endoperoxide Synthases / metabolism
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Prostaglandins / metabolism
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Transcription, Genetic / drug effects
Substances
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CEP-11004
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Carbazoles
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Enzyme Inhibitors
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Indoles
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Interleukin-6
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Isoenzymes
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NF-kappa B
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Prostaglandins
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3,9-bis((ethylthio)methyl)-K-252a
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Nitric Oxide
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Nitric Oxide Synthase
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Nitric Oxide Synthase Type II
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Nos2 protein, mouse
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Cyclooxygenase 2
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Prostaglandin-Endoperoxide Synthases
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JNK Mitogen-Activated Protein Kinases
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MAP Kinase Kinase Kinases
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MAP Kinase Kinase 4
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Mitogen-Activated Protein Kinase Kinases