Punicalagin inhibits neuroinflammation in LPS- activated rat primary microglia

Scope: In this study, the effects of punicalagin on neuroinflammation in LPS-activated microglia were investigated. Methods and results: The ability of punicalagin to reduce the production of TNF-alpha, IL-6 and prostaglandin E-2 was measured in culture medium using enzyme immunoassay. TNF-alpha and IL-6 gene expression in mouse hippocampal slices was measured with PCR. cyclooxygenase-2 and microsomal prostaglandin E synthase 1 protein and mRNA were evaluated with Western blotting and PCR, respectively. Further experiments to investigate effects of punicalagin on protein expressions of inflammatory targets were also determined with Western blotting. Pretreatment of rat primary microglia with punicalagin (5-40 mu M) prior to LPS (10 ng/mL) stimulation produced a significant (p < 0.05) inhibition of TNF-alpha, IL-6 and prostaglandin E-2 production. Punicalagin completely abolished TNF-alpha and IL-6 gene expression in LPS-stimulated hippocampal slices. Protein and mRNA expressions of cyclooxygenase-2 and microsomal prostaglandin E synthase 1 were also reduced by punicalagin pretreatment. Results show that punicalagin interferes with NF-kappa B signalling through attenuation of NF-kappa B-driven luciferase expression, as well as inhibition of I kappa B phosphorylation and nuclear translocation of p65 subunit in the microglia. Conclusion: These results suggest that punicalagin inhibits neuroinflammation in LPS-activated microglia through interference with NF-kappa B signalling, suggesting its potential as a nutritional preventive strategy in neurodegenerative disorders.