Increasing data from epidemiological and in vitro studies show that the isoflavonoids,
genistein and
daidzein, and the
flavonols,
quercetin and
kaempferol, are protective against
postmenopausal bone loss. However, the physiological mechanisms for these effects are not well understood. We now report that
kaempferol exerts profound antiosteoclastogenic effects by acting on both osteoblasts and osteoclasts.
Kaempferol but not
quercetin dose-dependently inhibited
tumor necrosis factor alpha (
TNFalpha)-induced production of the osteoclastogenic
cytokines interleukin-6 (IL-6) and
monocyte chemoattractant protein-1 (MCP-1/CCL2) in osteoblasts. The effect on
IL-6 was posttranscriptional, whereas
kaempferol reduced MCP-1
mRNA levels. In addition, in mouse primary calvarial osteoblasts,
kaempferol but not
quercetin blocked
TNFalpha-induced translocation of the
nuclear factor kappaB (
NF-kappaB) subunit p65 from the cytoplasm to the nucleus. However,
TNFalpha-stimulated intracellular ROS production was unaltered by
kaempferol. In RAW264.7 cells, a monocyte/macrophage precursor for osteoclasts, both
kaempferol and
quercetin dose-dependently inhibited the
receptor activator of NF-kappaB ligand (RANKL)-induced immediate-early oncogene c-fos expression at 6 h. After 3-5 days, both
flavonols robustly inhibited RANKL-induced expression of the osteoclastic
differentiation markers, RANK and
calcitonin receptor. Consistent with down regulation of these osteoclastic
differentiation markers, both
flavonols strongly attenuated the RANKL-induced formation of multinucleated osteoclasts. However,
kaempferol was more potent than
quercetin in inhibiting RANKL-stimulated effects on RAW264.7 cells. Thus, our data indicate that
kaempferol exerts profound antiosteoclastogenic effects by specifically antagonizing
TNF receptor family action on bone cells at two distinct levels, by disrupting production of osteoclastogenic
cytokines from osteoblasts and attenuating osteoclast precursor cell differentiation.