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Phytoestrogens directly inhibit TNF-α-induced bone resorption in RAW264.7 cells by suppressing c-fos-induced NFATc1
1School of Biomedical and Biological Sciences, University of Plymouth, Plymouth, UK.
Abstract:
TNF-α-induced osteoclastogenesis is central to post-menopausal and inflammatory bone loss, however, the effect of phytoestrogens on TNF-α-induced bone resorption has not been studied. The phytoestrogens genistein, daidzein, and coumestrol directly suppressed TNF-α-induced osteoclastogenesis and bone resorption. TRAP positive osteoclast formation and resorption area were significantly reduced by genistein (10(-7) M), daidzein (10(-5) M), and coumestrol (10(-7) M), which was prevented by the estrogen antagonist ICI 182,780. TRAP expression in mature TNF-α-induced osteoclasts was also significantly reduced by these phytoestrogen concentrations. In addition, in the presence of ICI 182,780 genistein and coumestrol (10(-5) -10(-6) M) augmented TNF-α-induced osteoclast formation and resorption. However, this effect was not observed in the absence of estrogen antagonist indicating that genistein's and coumestrol's ER-dependent anti-osteoclastic action normally negates this pro-osteoclastic effect. To determine the mechanism mediating the anti-osteoclastic action we examined the effect of genistein, coumestrol, and daidzein on caspase 3/7 activity, cell viability and expression of key genes regulating osteoclast differentiation and fusion. While anti-osteoclastic phytoestrogen concentrations had no effect on caspase 3/7 activity or cell viability they did significantly reduce TNF-α-induced c-fos and NFATc1 expression in an ER dependent manner and also inhibited NFATc1 nuclear translocation. Significant decreases in NFκB and DC-STAMP levels were also noted. Interestingly, constitutive c-fos expression prevented the anti-osteoclastic action of phytoestrogens on differentiation, resorption and NFATc1. This suggests that phytoestrogens suppress TNF-α-induced osteoclastogenesis via inhibition of c-fos-dependent NFATc1 expression. Our data provides further evidence that phytoestrogens have a potential role in the treatment of post-menopausal and inflammatory bone loss directly inhibiting TNF-α-induced resorption.
Insights
Phytoestrogens like genistein, daidzein, and coumestrol directly inhibit tumor necrosis factor-alpha (TNF-α)-induced osteoclastogenesis and bone resorption. This anti-osteoclastic action is mediated by suppressing key gene expression, offering potential for treating bone loss conditions.
Area of Science:
- Molecular Biology
- Endocrinology
- Bone Biology
Background:
- Tumor necrosis factor-alpha (TNF-α)-induced osteoclastogenesis drives post-menopausal and inflammatory bone loss.
- The impact of phytoestrogens on TNF-α-induced bone resorption remained uninvestigated.
Purpose of the Study:
- To investigate the direct effects of phytoestrogens (genistein, daidzein, coumestrol) on TNF-α-induced osteoclastogenesis and bone resorption.
- To elucidate the molecular mechanisms underlying the anti-osteoclastic actions of these phytoestrogens.
Main Methods:
- Assessed osteoclast formation and resorption in the presence of TNF-α and phytoestrogens.
- Utilized estrogen receptor antagonist (ICI 182,780) to determine the role of estrogen receptor (ER) dependency.
- Analyzed gene expression (c-fos, NFATc1, NFκB, DC-STAMP), nuclear translocation, caspase 3/7 activity, and cell viability.
Main Results:
- Genistein, daidzein, and coumestrol significantly suppressed TNF-α-induced osteoclastogenesis and bone resorption.
- These effects were ER-dependent and involved the inhibition of c-fos and NFATc1 expression and NFATc1 nuclear translocation.
- Phytoestrogens reduced NFκB and DC-STAMP levels, with constitutive c-fos expression blocking the anti-osteoclastic effects.
Conclusions:
- Phytoestrogens directly inhibit TNF-α-induced osteoclastogenesis and bone resorption through a c-fos-dependent suppression of NFATc1.
- These findings support a potential therapeutic role for phytoestrogens in managing post-menopausal and inflammatory bone loss.
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