LY294002 and LiCl Mitigate Neonatal ExPEC Meningitis Through Akt/GSK3β Signaling Modulation.

BACKGROUND

Neonatal purulent meningitis (NPM) is a life-threatening condition associated with high mortality rates and a substantial risk of severe long-term neurological sequelae among survivors. Extraintestinal pathogenic Escherichia coli (ExPEC) is the predominant causative agent of NPM and exhibits multidrug resistance. Targeting host signaling pathways is a promising therapeutic approach. In this study, we aimed to evaluate the therapeutic potential of LY294002 (a PI3K/Akt inhibitor) and lithium chloride (LiCl) (a GSK3β inhibitor) in a neonatal mouse model of ExPEC meningitis.

METHODS

Neonatal mice were infected with clinical ExPEC isolates to induce meningitis. LY294002 or LiCl was administered as an intervention. Brain bacterial load was quantified via culture, while inflammation was assessed using RT-qPCR to evaluate the expression of inflammatory cytokines; hematoxylin-eosin (HE) staining was also performed. The expression and localization of tight junction (TJ) proteins were analyzed using immunohistochemistry, and western blotting was used to quantify TJ proteins and key signaling molecules, including Akt, phosphorylated Akt (p-Akt Ser473), GSK3β, and phosphorylated GSK3β (p-GSK3β Ser9).

RESULTS

ExPEC colonization in the brain tissue was confirmed via bacterial culture. Early intervention with LiCl significantly reduced bacterial load. HE staining revealed meningeal thickening in infected mice, which was alleviated by both LY294002 and LiCl treatments. Western blotting and immunohistochemistry consistently demonstrated a marked reduction in the expression of TJ proteins following ExPEC infection, and their levels were substantially restored by both the interventions. These protective effects were associated with the modulation of the Akt/GSK3β signaling pathway.

CONCLUSION

LY294002 and LiCl reduced neuroinflammation and preserved the blood-brain barrier (BBB) integrity in the neonatal ExPEC meningitis model, likely via modulation of the Akt/GSK3β pathway. These results underscore the potential of this pathway as a therapeutic target and provide a basis for further preclinical investigation.