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Dual Effect of Chrysanthemum indicum Extract to Stimulate Osteoblast Differentiation and Inhibit Osteoclast Formation and Resorption In Vitro.

Baek JM, Kim JY, Cheon YH, Park SH, Ahn SJ, Yoon KH, Oh J, Lee MS - Evid Based Complement Alternat Med (2014)

Bottom Line: In the present study, we investigated the effect of Chrysanthemum indicum extract (CIE) on the differentiation of osteoclastic and osteoblastic cells.CIE strongly inhibited Akt, GSK3β, and IκB phosphorylation in RANKL-stimulated bone marrow macrophages and did not show any effects on MAP kinases, including p38, ERK, and JNK.Our results revealed that CIE could have a potential therapeutic role in bone-related disorders through its dual effects on osteoclast and osteoblast differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, School of Medicine, Wonkwang University, 344-2 Sinyong-dong, Iksan, Jeonbuk 570-749, Republic of Korea ; BK21plus Program & Department of Smart Life-Care Convergence, Graduate School, Wonkwang University, Iksan,Jeonbuk 570-749, Republic of Korea.

ABSTRACT
The risk of bone-related diseases increases due to the imbalance between bone resorption and bone formation by osteoclasts and osteoblasts, respectively. The goal in the development of antiosteoporotic treatments is an agent that will improve bone through simultaneous osteoblast stimulation and osteoclast inhibition without undesirable side effects. To achieve this goal, numerous studies have been performed to identify novel approaches using natural oriental herbs to treat bone metabolic diseases. In the present study, we investigated the effect of Chrysanthemum indicum extract (CIE) on the differentiation of osteoclastic and osteoblastic cells. CIE inhibited the formation of TRAP-positive mature osteoclasts and of filamentous-actin rings and disrupted the bone-resorbing activity of mature osteoclasts in a dose-dependent manner. CIE strongly inhibited Akt, GSK3β, and IκB phosphorylation in RANKL-stimulated bone marrow macrophages and did not show any effects on MAP kinases, including p38, ERK, and JNK. Interestingly, CIE also enhanced primary osteoblast differentiation via upregulation of the expression of alkaline phosphatase and the level of extracellular calcium concentrations during the early and terminal stages of differentiation, respectively. Our results revealed that CIE could have a potential therapeutic role in bone-related disorders through its dual effects on osteoclast and osteoblast differentiation.

No MeSH data available.


Related in: MedlinePlus

CIE downregulates RANKL-induced early signals and marker genes during osteoclastogenesis. (a) BMMs were pretreated with DMSO (control) or CIE (50 μg/mL) for 1 h in the presence of M-CSF (30 ng/mL) and were stimulated with RANKL (100 ng/mL) for the indicated times. Whole-cell lysates were used for western blot analysis with the specified antibodies. β-Actin served as the internal control. (b) BMMs were stimulated with RANKL (100 ng/mL) and M-CSF (30 ng/mL) in the presence or absence of CIE (50 μg/mL) for the indicated times. Total RNA was isolated from cells using QIAzol reagent and the mRNA expression levels of OSCAR, TRAP, integrin αv, β3, DC-STAMP, OC-STAMP, cathepsin K, and ICAM-1 were evaluated by real-time PCR.  *P < 0.05,  **P < 0.01, and  ***P < 0.001 versus the control at 12 h, 24 h, and 48 h.
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fig3: CIE downregulates RANKL-induced early signals and marker genes during osteoclastogenesis. (a) BMMs were pretreated with DMSO (control) or CIE (50 μg/mL) for 1 h in the presence of M-CSF (30 ng/mL) and were stimulated with RANKL (100 ng/mL) for the indicated times. Whole-cell lysates were used for western blot analysis with the specified antibodies. β-Actin served as the internal control. (b) BMMs were stimulated with RANKL (100 ng/mL) and M-CSF (30 ng/mL) in the presence or absence of CIE (50 μg/mL) for the indicated times. Total RNA was isolated from cells using QIAzol reagent and the mRNA expression levels of OSCAR, TRAP, integrin αv, β3, DC-STAMP, OC-STAMP, cathepsin K, and ICAM-1 were evaluated by real-time PCR.  *P < 0.05,  **P < 0.01, and  ***P < 0.001 versus the control at 12 h, 24 h, and 48 h.

Mentions: To elucidate the inhibitory mechanism and pathways influenced by CIE, we evaluated the effect of CIE on MAP kinases, including p38, ERK, JNK, and IκB, Akt, and the GSK3β pathway, which are widely known to be essential for osteoclast differentiation. As shown in Figure 3(a), RANKL-induced phosphorylation of MAP kinases was not affected by treatment with CIE. However, the phosphorylation of Akt, GSK3β, and IκB and the degradation of IκB by RANKL were decreased by treatment with CIE. In addition, we also analysed the effect of CIE on various genes related to osteoclast formation and function at the mRNA level. CIE downregulated the expression of OSCAR and TRAP, which are genes specifically related to osteoclast formation. Moreover, CIE also suppressed DC-STAMP, OC-STAMP, ICAM-1, and integrin αvβ3 expression, which are known to affect cell-cell interactions such as migration or fusion. Expression of cathepsin K, which is related to bone-resorbing activity, was also significantly inhibited (Figure 3(b)). These results indicate that the inhibitory effects of CIE on RANKL-mediated NF-κB, Akt, and GSK3β signalling are followed by the inactivation of osteoclast-specific marker genes.


Dual Effect of Chrysanthemum indicum Extract to Stimulate Osteoblast Differentiation and Inhibit Osteoclast Formation and Resorption In Vitro.

Baek JM, Kim JY, Cheon YH, Park SH, Ahn SJ, Yoon KH, Oh J, Lee MS - Evid Based Complement Alternat Med (2014)

CIE downregulates RANKL-induced early signals and marker genes during osteoclastogenesis. (a) BMMs were pretreated with DMSO (control) or CIE (50 μg/mL) for 1 h in the presence of M-CSF (30 ng/mL) and were stimulated with RANKL (100 ng/mL) for the indicated times. Whole-cell lysates were used for western blot analysis with the specified antibodies. β-Actin served as the internal control. (b) BMMs were stimulated with RANKL (100 ng/mL) and M-CSF (30 ng/mL) in the presence or absence of CIE (50 μg/mL) for the indicated times. Total RNA was isolated from cells using QIAzol reagent and the mRNA expression levels of OSCAR, TRAP, integrin αv, β3, DC-STAMP, OC-STAMP, cathepsin K, and ICAM-1 were evaluated by real-time PCR.  *P < 0.05,  **P < 0.01, and  ***P < 0.001 versus the control at 12 h, 24 h, and 48 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: CIE downregulates RANKL-induced early signals and marker genes during osteoclastogenesis. (a) BMMs were pretreated with DMSO (control) or CIE (50 μg/mL) for 1 h in the presence of M-CSF (30 ng/mL) and were stimulated with RANKL (100 ng/mL) for the indicated times. Whole-cell lysates were used for western blot analysis with the specified antibodies. β-Actin served as the internal control. (b) BMMs were stimulated with RANKL (100 ng/mL) and M-CSF (30 ng/mL) in the presence or absence of CIE (50 μg/mL) for the indicated times. Total RNA was isolated from cells using QIAzol reagent and the mRNA expression levels of OSCAR, TRAP, integrin αv, β3, DC-STAMP, OC-STAMP, cathepsin K, and ICAM-1 were evaluated by real-time PCR.  *P < 0.05,  **P < 0.01, and  ***P < 0.001 versus the control at 12 h, 24 h, and 48 h.
Mentions: To elucidate the inhibitory mechanism and pathways influenced by CIE, we evaluated the effect of CIE on MAP kinases, including p38, ERK, JNK, and IκB, Akt, and the GSK3β pathway, which are widely known to be essential for osteoclast differentiation. As shown in Figure 3(a), RANKL-induced phosphorylation of MAP kinases was not affected by treatment with CIE. However, the phosphorylation of Akt, GSK3β, and IκB and the degradation of IκB by RANKL were decreased by treatment with CIE. In addition, we also analysed the effect of CIE on various genes related to osteoclast formation and function at the mRNA level. CIE downregulated the expression of OSCAR and TRAP, which are genes specifically related to osteoclast formation. Moreover, CIE also suppressed DC-STAMP, OC-STAMP, ICAM-1, and integrin αvβ3 expression, which are known to affect cell-cell interactions such as migration or fusion. Expression of cathepsin K, which is related to bone-resorbing activity, was also significantly inhibited (Figure 3(b)). These results indicate that the inhibitory effects of CIE on RANKL-mediated NF-κB, Akt, and GSK3β signalling are followed by the inactivation of osteoclast-specific marker genes.

Bottom Line: In the present study, we investigated the effect of Chrysanthemum indicum extract (CIE) on the differentiation of osteoclastic and osteoblastic cells.CIE strongly inhibited Akt, GSK3β, and IκB phosphorylation in RANKL-stimulated bone marrow macrophages and did not show any effects on MAP kinases, including p38, ERK, and JNK.Our results revealed that CIE could have a potential therapeutic role in bone-related disorders through its dual effects on osteoclast and osteoblast differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, School of Medicine, Wonkwang University, 344-2 Sinyong-dong, Iksan, Jeonbuk 570-749, Republic of Korea ; BK21plus Program & Department of Smart Life-Care Convergence, Graduate School, Wonkwang University, Iksan,Jeonbuk 570-749, Republic of Korea.

ABSTRACT
The risk of bone-related diseases increases due to the imbalance between bone resorption and bone formation by osteoclasts and osteoblasts, respectively. The goal in the development of antiosteoporotic treatments is an agent that will improve bone through simultaneous osteoblast stimulation and osteoclast inhibition without undesirable side effects. To achieve this goal, numerous studies have been performed to identify novel approaches using natural oriental herbs to treat bone metabolic diseases. In the present study, we investigated the effect of Chrysanthemum indicum extract (CIE) on the differentiation of osteoclastic and osteoblastic cells. CIE inhibited the formation of TRAP-positive mature osteoclasts and of filamentous-actin rings and disrupted the bone-resorbing activity of mature osteoclasts in a dose-dependent manner. CIE strongly inhibited Akt, GSK3β, and IκB phosphorylation in RANKL-stimulated bone marrow macrophages and did not show any effects on MAP kinases, including p38, ERK, and JNK. Interestingly, CIE also enhanced primary osteoblast differentiation via upregulation of the expression of alkaline phosphatase and the level of extracellular calcium concentrations during the early and terminal stages of differentiation, respectively. Our results revealed that CIE could have a potential therapeutic role in bone-related disorders through its dual effects on osteoclast and osteoblast differentiation.

No MeSH data available.


Related in: MedlinePlus