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MST4 Regulates Microglia Neuroinflammation via Targeting PKM2 Nuclear Translocation in Epilepsy: An In Vivo and In
Jie Fu1, Shaotao Zhang2, Yifei Chu3
1Department of Neurology, the Affiliated Hospital of Southwest Medical University, Taiping Street, Jiangyang District, Luzhou, 646000, China. fujessie@swmu.edu.cn.
Abstract:
Microglia-mediated neuroinflammation is closely associated with the pathogenesis of epilepsy. Mammalian sterile-20-like kinase 4 (MST4) has been suggested a regulator of inflammation. However, the effect of MST4 on microglia neuroinflammation in epilepsy remains unclear. A pilocarpine-induced rat epilepsy model was constructed and a lipopolysaccharide (LPS)-stimulated microglia cell model was applied in the current research. Knockdown or overexpression of MST4 was established using lentivirus transfection. Electroencephalograph (EEG) was employed to measure brain activities of rats. The protein and mRNA expressions were detected using western blot and qRT-PCR, respectively. Immunofluorescent staining was conducted to detect the distribution of the proteins. TUNEL staining was performed to evaluate cell apoptosis. The protein interaction was evaluated with Co-IP assay. Our results showed that MST4 and nuclear PKM2 expressions were increased in epileptic rats compared to control and colocalized with microglia. MST4 overexpression inhibited microglia activation, the release of TNF-α and IL-1β, and improved neuronal apoptosis in epileptic rats. Furthermore, MST4 interacted with PKM2 and regulated PKM2 nuclear translocation. Inhibiting PKM2 nuclear translocation by TEPP-46 reversed the promoting effect of MST4 knockdown on microglia neuroinflammation. In summary, our study demonstrated that MST4 alleviated microglia-mediated neuroinflammation in epilepsy, and the mechanism of MST4-mediated anti-neuroinflammatory effects may be associated with the inhibition of PKM2 nuclear translocation.
Insights
Mammalian sterile-20-like kinase 4 (MST4) reduces neuroinflammation in epilepsy by inhibiting PKM2 nuclear translocation. This finding offers a potential therapeutic target for epilepsy treatment.
Area of Science:
- Neuroscience
- Inflammation Research
- Epilepsy Pathogenesis
Background:
- Microglia-driven neuroinflammation is a key factor in epilepsy development.
- Mammalian sterile-20-like kinase 4 (MST4) is implicated in inflammatory processes, but its role in epilepsy-related neuroinflammation is unknown.
Purpose of the Study:
- To investigate the effect of MST4 on microglia-mediated neuroinflammation in epilepsy.
- To elucidate the underlying molecular mechanisms involving MST4 in epilepsy.
Main Methods:
- Established a pilocarpine-induced rat epilepsy model and lipopolysaccharide (LPS)-stimulated microglia model.
- Utilized lentivirus for MST4 knockdown/overexpression, EEG for brain activity, western blot/qRT-PCR for gene/protein expression, immunofluorescence, TUNEL staining, and Co-IP assay.
- Investigated the interaction between MST4 and PKM2 and its effect on PKM2 nuclear translocation.
Main Results:
- MST4 and nuclear PKM2 expression increased in epileptic rats and co-localized with microglia.
- MST4 overexpression suppressed microglia activation, reduced TNF-α and IL-1β release, and decreased neuronal apoptosis.
- MST4 interacted with PKM2, inhibiting its nuclear translocation; blocking PKM2 nuclear translocation reversed MST4 knockdown-induced neuroinflammation.
Conclusions:
- MST4 alleviates microglia-mediated neuroinflammation in epilepsy.
- MST4's anti-neuroinflammatory mechanism involves the inhibition of PKM2 nuclear translocation.
