Activation of the liver X receptor-ß potently inhibits osteoclastogenesis from lipopolysaccharide-exposed bone marrow-derived macrophages.

Bacterial-induced bone diseases, such as periodontitis and osteomyelitis, are chronic inflammatory diseases characterized by increased bone destruction as a result of enhanced osteoclastogenesis. The LXRα and -ß are important modulators of inflammatory signaling and can potently inhibit RANKL-induced osteoclast differentiation. Here, we investigated the effects of the LXR agonist GW3965 on LPS-induced osteoclast differentiation. Mouse BMMs primed with RANKL for 24 h, then exposed to LPS in the presence of GW3965 for 4 days, formed significantly fewer and smaller TRAP(+)-multinucleated osteoclasts with reduced expression of osteoclast markers (Acp5, Ctsk, Mmp-9, Dc-stamp, and Itgß3), along with inhibition of actin ring development. GW3965 was able to repress proinflammatory cytokine (TNF-α, IL-1ß, IL-6, and IL-12p40) expression in BMMs exposed to LPS alone; however, once BMMs entered the osteoclast lineage following RANKL priming, GW3965 no longer inhibited cytokine expression. The inhibitory action of GW3965 involved the Akt pathway but seemed to be independent of MAPKs (p38, ERK, JNK) and NF-κB signaling. GW3965 acted in a LXRß-dependent mechanism, as osteoclast differentiation was not inhibited in BMMs derived from LXRß-/- mice. Finally, activation of LXR also inhibited differentiation in LPS-exposed mouse RAW264.7 cells. In conclusion, GW3965 acts through LXRß to potently inhibit osteoclast differentiation from RANKL-primed BMMs in a LPS environment. In this respect, activation of the LXR could have a beneficial, therapeutic effect in the prevention of bacterial-induced bone erosion.

The liver X receptor promotes macrophage differentiation and suppresses osteoclast formation in mouse RAW264.7 promyelocytic leukemia cells exposed to bacterial lipopolysaccharide.

Lipopolysaccharide (LPS), the principal component of Gram-negative bacterial cell walls, is a stimulator of osteoclastogenesis and thus a key factor in inflammatory bone loss. We have recently reported that the important cholesterol and inflammatory regulator, liver X receptor (LXRα/ß), can potently inhibit osteoclast formation from bone marrow-derived osteoclast precursors in a bacterial/LPS environment. In this manuscript, we further studied the effect of the LXR agonist GW3965 on osteoclast differentiation in RAW264.7 promyelocytic leukemia cells exposed to LPS. We found that not only did activation of the LXR potently inhibit the formation of TRAP-positive osteoclast-like cells, but promoted a population of TRAP-negative mononuclear cells with high phagocytic activity. We observed reduced expression of the osteoclast markers TRAP/Acp5, Ctsk, Calcr and Oscar after 3-4days of GW3965 treatment, coinciding with an increase in the expression of the anti-osteoclastogenic factor Irf8. Expression of the macrophage/phagocytic marker Cd68 was increased, however the "classical" macrophage markers F4/80 and Cd14 and the "alternatively" activated macrophage markers Tgfß and Il10 were not altered. Further, activation of LXR increased the expression of the macrophage survival gene AIM/SPα, a known LXR target gene, and osteoclast/macrophage-related markers (Mitf, Pu.1, Usf1/2, Ostm1 and Mfr). Although Akt phosphorylation was reduced, GW3965 seemed to act independently of MAPKs (p38, ERK, JNK) and NFκB, and had no inhibitory effect on cytokine expression (Tnfα, Il6, or Il1ß). Our results indicate that activation of the LXR not only inhibits the differentiation of osteoclast-like cells from RAW264.7 cells in a bacterial/LPS environment, but is also involved in the fate determination of myeloid progenitor cells into macrophages with high phagocytic capacity.