Limits...
Inhibition of Osteoclastogenesis and Bone Resorption in vitro and in vivo by a prenylflavonoid xanthohumol from hops.

Li J, Zeng L, Xie J, Yue Z, Deng H, Ma X, Zheng C, Wu X, Luo J, Liu M - Sci Rep (2015)

Bottom Line: In this study, we examined the effects of xanthohumol (XN), an abundant prenylflavonoid from hops plant, on osteoclastogenesis, osteoclast resorption, and RANKL-induced signaling pathway using both in vitro and in vivo assay systems.At the molecular level, XN disrupted the association of RANK and TRAF6, resulted in the inhibition of NF-κB and Ca(2+)/NFATc1 signaling pathway during osteoclastogenesis.As a results, XN suppressed the expression of osteoclastogenesis-related marker genes, including CtsK, Nfatc1, Trap, Ctr.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Fengxian District Central Hospital and East China Normal University Joint Center for Translational Medicine, Shanghai Key laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai 200241, China.

ABSTRACT
Excessive RANKL signaling leads to superfluous osteoclast formation and bone resorption, is widespread in the pathologic bone loss and destruction. Therefore, targeting RANKL or its signaling pathway has been a promising and successful strategy for this osteoclast-related diseases. In this study, we examined the effects of xanthohumol (XN), an abundant prenylflavonoid from hops plant, on osteoclastogenesis, osteoclast resorption, and RANKL-induced signaling pathway using both in vitro and in vivo assay systems. In mouse and human, XN inhibited osteoclast differentiation and osteoclast formation at the early stage. Furthermore, XN inhibited osteoclast actin-ring formation and bone resorption in a dose-dependent manner. In ovariectomized-induced bone loss mouse model and RANKL-injection-induced bone resorption model, we found that administration of XN markedly inhibited bone loss and resorption by suppressing osteoclast activity. At the molecular level, XN disrupted the association of RANK and TRAF6, resulted in the inhibition of NF-κB and Ca(2+)/NFATc1 signaling pathway during osteoclastogenesis. As a results, XN suppressed the expression of osteoclastogenesis-related marker genes, including CtsK, Nfatc1, Trap, Ctr. Therefore, our data demonstrated that XN inhibits osteoclastogenesis and bone resorption through RANK/TRAF6 signaling pathways. XN could be a promising drug candidate in the treatment of osteoclast-related diseases such as postmenopausal osteoporosis.

No MeSH data available.


Related in: MedlinePlus

XN represses the association of RANK and TRAF6, and abrogates osteoclastogenesis-related gene expression.(A,B) XN suppressed the RANKL-induced binding of RANK and TRAF6. The RAW264.7 cells were pretreated with indicated concentration of XN for 4 hours then stimulated with RANKL (30 ng/ml) for another 20 minutes. The cell lysate were harvest and immunoprecipitated with antibody to RANK and then blotted with anti-TRAF6 (A). Or the cell lysates were immunoprecipitated with antibody to TRAF6 and then blotted with anti-RANK (B). (C) XN inhibits the mRNA levels of NFATc1, cathepsin K (CtsK), CTR and TRAP induced by RANKL. Mouse BMMs were incubated with XN (5 μM) and RANKL (30 ng/mg) for indicate day. Total RNA was collected and analyzed by Real time-PCR. Column, means of experiments performed in triplicate; bar, SD. **p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4664947&req=5

f7: XN represses the association of RANK and TRAF6, and abrogates osteoclastogenesis-related gene expression.(A,B) XN suppressed the RANKL-induced binding of RANK and TRAF6. The RAW264.7 cells were pretreated with indicated concentration of XN for 4 hours then stimulated with RANKL (30 ng/ml) for another 20 minutes. The cell lysate were harvest and immunoprecipitated with antibody to RANK and then blotted with anti-TRAF6 (A). Or the cell lysates were immunoprecipitated with antibody to TRAF6 and then blotted with anti-RANK (B). (C) XN inhibits the mRNA levels of NFATc1, cathepsin K (CtsK), CTR and TRAP induced by RANKL. Mouse BMMs were incubated with XN (5 μM) and RANKL (30 ng/mg) for indicate day. Total RNA was collected and analyzed by Real time-PCR. Column, means of experiments performed in triplicate; bar, SD. **p < 0.01.

Mentions: Since the NF-κB and Ca2+/NFATc1 pathways share the same upstream molecule, TRAF6, which is associated with RANK when activated by RANKL28, we then examined the possible effect of XN in the binding between RANK and TRAF6. In the endogenous coimmunoprecipitation assay, XN suppressed RANKL-induced association of RANK and TRAF6 in a dose-dependent manner when immunoprecipitated with antibody to RANK and blotted with anti-TRAF6 (Fig. 7A). On the other end, XN also inhibited the binding when immunoprecipitated with antibody to TRAF6 and blotted with anti-RANK (Fig. 7B). All of the results suggested that XN could dissociate the signal complex RANK and TRAF6.


Inhibition of Osteoclastogenesis and Bone Resorption in vitro and in vivo by a prenylflavonoid xanthohumol from hops.

Li J, Zeng L, Xie J, Yue Z, Deng H, Ma X, Zheng C, Wu X, Luo J, Liu M - Sci Rep (2015)

XN represses the association of RANK and TRAF6, and abrogates osteoclastogenesis-related gene expression.(A,B) XN suppressed the RANKL-induced binding of RANK and TRAF6. The RAW264.7 cells were pretreated with indicated concentration of XN for 4 hours then stimulated with RANKL (30 ng/ml) for another 20 minutes. The cell lysate were harvest and immunoprecipitated with antibody to RANK and then blotted with anti-TRAF6 (A). Or the cell lysates were immunoprecipitated with antibody to TRAF6 and then blotted with anti-RANK (B). (C) XN inhibits the mRNA levels of NFATc1, cathepsin K (CtsK), CTR and TRAP induced by RANKL. Mouse BMMs were incubated with XN (5 μM) and RANKL (30 ng/mg) for indicate day. Total RNA was collected and analyzed by Real time-PCR. Column, means of experiments performed in triplicate; bar, SD. **p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664947&req=5

f7: XN represses the association of RANK and TRAF6, and abrogates osteoclastogenesis-related gene expression.(A,B) XN suppressed the RANKL-induced binding of RANK and TRAF6. The RAW264.7 cells were pretreated with indicated concentration of XN for 4 hours then stimulated with RANKL (30 ng/ml) for another 20 minutes. The cell lysate were harvest and immunoprecipitated with antibody to RANK and then blotted with anti-TRAF6 (A). Or the cell lysates were immunoprecipitated with antibody to TRAF6 and then blotted with anti-RANK (B). (C) XN inhibits the mRNA levels of NFATc1, cathepsin K (CtsK), CTR and TRAP induced by RANKL. Mouse BMMs were incubated with XN (5 μM) and RANKL (30 ng/mg) for indicate day. Total RNA was collected and analyzed by Real time-PCR. Column, means of experiments performed in triplicate; bar, SD. **p < 0.01.
Mentions: Since the NF-κB and Ca2+/NFATc1 pathways share the same upstream molecule, TRAF6, which is associated with RANK when activated by RANKL28, we then examined the possible effect of XN in the binding between RANK and TRAF6. In the endogenous coimmunoprecipitation assay, XN suppressed RANKL-induced association of RANK and TRAF6 in a dose-dependent manner when immunoprecipitated with antibody to RANK and blotted with anti-TRAF6 (Fig. 7A). On the other end, XN also inhibited the binding when immunoprecipitated with antibody to TRAF6 and blotted with anti-RANK (Fig. 7B). All of the results suggested that XN could dissociate the signal complex RANK and TRAF6.

Bottom Line: In this study, we examined the effects of xanthohumol (XN), an abundant prenylflavonoid from hops plant, on osteoclastogenesis, osteoclast resorption, and RANKL-induced signaling pathway using both in vitro and in vivo assay systems.At the molecular level, XN disrupted the association of RANK and TRAF6, resulted in the inhibition of NF-κB and Ca(2+)/NFATc1 signaling pathway during osteoclastogenesis.As a results, XN suppressed the expression of osteoclastogenesis-related marker genes, including CtsK, Nfatc1, Trap, Ctr.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Fengxian District Central Hospital and East China Normal University Joint Center for Translational Medicine, Shanghai Key laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai 200241, China.

ABSTRACT
Excessive RANKL signaling leads to superfluous osteoclast formation and bone resorption, is widespread in the pathologic bone loss and destruction. Therefore, targeting RANKL or its signaling pathway has been a promising and successful strategy for this osteoclast-related diseases. In this study, we examined the effects of xanthohumol (XN), an abundant prenylflavonoid from hops plant, on osteoclastogenesis, osteoclast resorption, and RANKL-induced signaling pathway using both in vitro and in vivo assay systems. In mouse and human, XN inhibited osteoclast differentiation and osteoclast formation at the early stage. Furthermore, XN inhibited osteoclast actin-ring formation and bone resorption in a dose-dependent manner. In ovariectomized-induced bone loss mouse model and RANKL-injection-induced bone resorption model, we found that administration of XN markedly inhibited bone loss and resorption by suppressing osteoclast activity. At the molecular level, XN disrupted the association of RANK and TRAF6, resulted in the inhibition of NF-κB and Ca(2+)/NFATc1 signaling pathway during osteoclastogenesis. As a results, XN suppressed the expression of osteoclastogenesis-related marker genes, including CtsK, Nfatc1, Trap, Ctr. Therefore, our data demonstrated that XN inhibits osteoclastogenesis and bone resorption through RANK/TRAF6 signaling pathways. XN could be a promising drug candidate in the treatment of osteoclast-related diseases such as postmenopausal osteoporosis.

No MeSH data available.


Related in: MedlinePlus