Cysteinyl Leukotriene Receptor 2 is Involved in Inflammation and Neuronal Damage by Mediating Microglia M1/M2 Polarization through NF-κB Pathway

Abstract

Microglia activation plays a key role in regulating inflammatory and immune reaction during cerebral ischemia and it exerts pro-inflammatory or anti-inflammatory effect depending on M₁/M₂ polarization phenotype. Cysteinyl leukotriene 2 receptor (CysLT₂R) is a potent inflammatory mediator receptor, and involved in cerebral ischemic injury, but the mechanism of CysLT₂R regulating inflammation and neuron damage remains unclear. Here, we found that LPS and CysLT₂R agonist NMLTC₄ significantly increased microglia proliferation and phagocytosis, up-regulated the mRNA expression of M₁ polarization markers (IL-1β, TNF-α, IFN-γ, CD86 and iNOS), down-regulated the expression of M₂ polarization markers (Arg-1, CD206, TGF-β, IL-10, Ym-1) and increased the release of IL-1β and TNF-α. CysLT₂R selective antagonist HAMI3379 could antagonize these effects. IL-4 significantly up-regulated the mRNA expression of M₂ polarization markers, and HAMI3379 further increased IL-4-induced up-regulation of M₂ polarization markers expression. Additionally, LPS and NMLTC₄ stimulated NF-κB p50 and p65 proteins expression, and promoted p50 transfer to the nucleus. Pre-treatment with HAMI3379 and NF-κB signaling inhibitor Bay 11-7082 could reverse the up-regulation of p50 and p65 proteins expression, and inhibited p50 transfer to the nucleus. The conditional medium of BV-2 cells contained HAMI3379 could inhibit SH-SY5Y cells apoptosis induced by LPS and NMLTC₄. These results were further confirmed in primary microglia. The findings indicate that CysLT₂R was involved in inflammation and neuronal damage by inducing the activation of microglia M1 polarization and NF-κB pathway, inhibiting microglia M1 polarization and promoting microglia polarization toward M2 phenotype which may exerts neuroprotective effects, and targeting CysLT₂R may be a new therapeutic strategy against cerebral ischemia stroke.

Keywords: CysLT(2)R; M1/M2 polarization; NF-κB pathway; inflammation; neurons damage.