Inhibition of the TLR4/NF-κB pathway promotes the polarization of LPS-induced BV2 microglia toward the M2 phenotype

This study aimed to investigate whether the inhibition of the TLR4/NF-kappa B pathway can promote lipopolysaccharide (LPS)-induced microglial polarization from the M1 to M2 phenotype, and thus exert neuroprotection. LPS-induced microglia were used as a model for inflammation in vitro. TLR4-specific inhibitor resatorvid (TAK-242) and NF-kappa B inhibitor pyrrolidine dithiocarbamate (PDTC) were used to verify the effect of the TLR4/NF-kappa B pathway on microglia activation and polarization. Cell proliferation was measured by cell counting, and nitric oxide (NO) and reactive oxygen species (ROS) release was measured using the Griess reagent and ROS kit, respectively. Immunofluorescence and RT-qPCR analyses were used to detect the expression of microglial activation markers, phenotypic markers, related pathway molecules, and inflammatory factors. TLR4 specific inhibitor TAK-242 and NF-kappa B inhibitor PDTC alleviated LPS-induced microglia over-activation by inhibiting the TLR4/NF-kappa B pathway, and reduced LPS-stimulated cell proliferation and the release of NO, ROS, TNF-a, and IL-6 and IL-1 beta. Meanwhile, TAK-242 and PDTC promoted LPS-induced polarization of microglia from M1 to M2 phenotype, decreased the expression of microglial activation marker Iba1 and M1 phenotypic markers (TNF-a and CD86), and increased the expression of M2 phenotypic markers (Arg-1 and CD206). The mechanism may be related to inhibiting the TLR4/NF-kappa B pathway. The inhibition of the TLR4/NF-kappa B pathway can promote LPS-induced polarization of BV2 microglia from M1 phenotype to M2 phenotype.

Automated summary

This study found that inhibiting the TLR4/NF-κB pathway can promote the polarization of LPS-induced microglia from M1 to M2 phenotype and alleviate excessive cell activation and inflammatory factor release.