• The Journal of Korean Obstetrics and Gynecology
• /
• v.27 no.3
• /
• pp.12-27
• /
• 2014

Objectives: This study was performed to evaluate the effect of Guibi-tang water extract (GB) on osteoporosis. Methods: We examined the effect of GB on osteoclast differentiation using murine pre-osteoclastic RAW 264.7 cells treated with receptor activator of nuclear factor kappa-B ligand (RANKL). The effect of GB on osteoclast was measured by counting TRAP (+) multinucleated cells and measuring TRAP activity. The mRNA expressions of osteoclastogenesis-related genes (Cathepsin K, MMP-9, TRAP, NFATc1, MITF, TNF-${\alpha}$, IL-6, COX-2) were measured by real-time PCR. We examined the effect of GB on osteoblast proliferation, ALP activity, bone matrix protein synthesis and collagen synthesis using murine calvarial cell. Results: GB decreased the number of TRAP (+) multinucleated cells and inhibited TRAP activity in RANKL-stimulated RAW 264.7 cell. GB decreased the expression of genes related osteoclastogenesis such as Cathepsin K, MMP-9, TRAP, NFATc1, MITF, COX-2 in RANKL-stimulated RAW 264.7 cell. But GB did not decrease the expression of iNOS and increased the expression of TNF-${\alpha}$, IL-6 in RANKL-stimulated RAW 264.7 cell. These genes (iNOS, TNF-${\alpha}$, IL-6) are thought to be related with the inflammatory bone destruction. GB increased cell proliferation of rat calvarial cell and also increased ALP activity in rat calvarial cell. GB did not increase bone matrix protein synthesis but increased collagen synthesis in rat calvarial cell. Conclusions: This study suggests that GB may be effective in treating osteoporosis by inhibiting osteoclast differentiation and its related gene expression and by increasing osteoblast proliferation.

• Journal of Periodontal and Implant Science
• /
• v.29 no.3
• /
• pp.645-654
• /
• 1999

Interleukin-6(IL-6) stimulate osteoclast differentiation. $17{\beta}$-estradiol, 1,25-dihydroxyvitamin $D_3$(1,25-$(OH)_2D_3$) and interleukin-$1{\beta}$ inhibit or stimulate osteoclast differentiation by decreasing or increasing the synthesis of interleukin-6(IL-6) from stromal/osteoblastic cells, respectively. Periodontal ligament(PDL) cells reside between the alveolar bone and the cementum and have osteoblastic characteristics. To estimate the effect of $17{\beta}$-estradiol and 1,25$(OH)_2D_3$ on IL-6 production of PDL cells, PDL cells were treated with $17{\beta}$-estradiol or 1,25-$(OH)_2D_3$ in the absence or the presence of IL-$1{\beta}$. The concentration of IL-6 produced form PDL cells was determined by enzym linked immunosorbent assay(ELISA). In unstimulated PDL cells, we detected constitutive production of IL-6 at 1st and 2nd day. IL-$1{\beta}$ increased IL-6 synthesis at 1st day and 2nd day. $17{\beta}$-estradiol had no significant effect on the secretion of this cytokine, either constitutively or after stimulation with IL- $1{\beta}$(0.05 ng/ml). 1,25-$(OH)_2D_3$($10^{-8}M$) decreased not only constitutive IL-6 production but also IL-$1{\beta}$-induced IL-6 production at 2nd day. These results suggest that 1,25-$(OH)_2D_3$ may control IL-$1{\beta}$-induced osteoclast differentiation by decreasing IL-$1{\beta}$-induced IL-6 secretion of PDL cells.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.23 no.4
• /
• pp.860-865
• /
• 2009

Osteoclasts are bone-resorbing multinucleated cells derived from the monocyte/macrophage lineage. The differentiation of osteoclasts are regulated by osteoblastic cells expressed RANKL, which is the most critical molecule for osteoclast differentiation. In this study, we found that water extracts of cuscuta inhibited RANKL-mediated osteoclast differentiation by direct action on bone marrow macrophages (BMMs) without cytotoxicity. In BMMs, water extracts of cuscuta inhibited the mRNA expression of c-Fos, NFATc1, TRAP, and OSCAR. Also, the protein expression of c-Fos and NFATc1 was inhibited by water extracts of cuscuta treatement. Water extracts of cuscuta inhibited the phosphorylation of p38, ERK, and JNK in BMMs treated with RANKL. However, water extracts of cuscuta did not inhibit RANKL-induced I-${\kappa}B$ activation. Water extract of cuscuta failed to inhibit bone resorption by osteoclasts cultured on hydroxyapatite plates. These results suggest that cuscuta may be a promising drug for use against bone disorders such as osteoporosis and rheumatoid arthritis.

• Herbal Formula Science
• /
• v.18 no.2
• /
• pp.167-182
• /
• 2010

Cervi parvum cornu (Deer Antler) and Carthami tinctorii fructus (Also known as Carthami seed) are widely used for treating osteoporosis and rheumatoid arthritis. In this study, We found out that the water extract of Cervi parvum cornu(WECPC), Carthami tinctorii fructus(WECTF) and their combination have effects of suppressing the RANKL-induced osteoclast differentiation. We assayed mRNA expression levels of NFATc1, c-Fos, TRAP and GAPDHS from bone marrow macrophages(BMMs) by means of RT-PCR. Similarly, the protein expression levels of NFATc1, c-Fos, MAPKs and $\beta$-actin in cell lysates were analyzed by means of Western blotting. then we determined the anti-osteoporotic effects of WECPC, WECTF and their combination using Lipopolysaccharide (LPS)-induced bone-loss mouse. WECPC, WECTF and their combination showed remarkable inhibition on RANKL-treated osteoclast differentiation without cytotoxicity. WECPC suppressed degradation of I-${\kappa}B$. WECPC, WECTF and their combination down-regulated the induction of c-Fos and NFATc1 by RANKL. Lastly, in vivo data showed that WECPC, WECTF and their combination rescued the bone erosion by LPS treatment. Thus, these results demonstrate that WECPC, WECTF and their combination can be efficacious remedies for bone-loss diseases such as osteoporosis and rheumatoid arthritis.

• The Korea Journal of Herbology
• /
• v.25 no.1
• /
• pp.65-74
• /
• 2010

Objectives : Achyranthis Bidentatae Radix (ABR) has been used for treating of many symptoms especially osteoporosis and rheumatoid arthritis. In this study, we determined the effects of water extract of ABR in RANKL (Receptor Activator for Nuclear Factor $\kappa$ B Ligand)-induced osteoclast differentiation culture system. Methods : We assayed mRNA expression levels of NFATc1, c-Fos, TRAP, OSCAR, $FcR{\gamma}$, DAP12 and GAPDH in bone marrow macrophages (BMMs) treated with ABR. The protein expression levels of NFATc1, c-Fos, MAPKs and $\beta$-actin in cell lysates treated with ABR were analysed by Western blotting. In addition we determined the effects of water extract of ABR on LPS-induced bone-loss mouse. Results : Water extract of ABR showed remarkable inhibition on RANKL-treated osteoclast differentiation without cytotoxicity. ABR down-regulated the induction of c-Fos and NFATc1 by RANKL. ABR suppressed phosphorylation of JNK, p38 and I-${\kappa}B$. ABR rescued bone erosion by LPS induction in vivo study. Conclusions : These results demonstrate that ABR may be a useful remedy for curing of bone-loss disease such as osteoporosis.

• Preventive Nutrition and Food Science
• /
• v.19 no.4
• /
• pp.353-357
• /
• 2014

Phytic acid (myo-inositol hexakisphosphate) or phytate is considered an anti-nutrient due to the formation of precipitated complexes that strongly reduces the absorption of essential dietary minerals. In this study, brown rice with reduced phytate was made by inoculation with Lactobacillus sakei Wikim001 having high phytase activity. The effects of brown rice extract treated with L. sakei Wikim001 (BR-WK) on osteoblast differentiation and osteoclast formation were investigated. The proliferation of SaOS-2 cells was measured by the MTT assay. Treatment with BR-WK increased cell proliferation by 136% at a concentration of $100{\mu}g/mL$. The Alkaline phosphate activity in SaOS-2 cells was 129% higher when BR-WK was processed at a concentration of $100{\mu}g/mL$. The proliferation of bone marrow macrophages decreased by nearly 60% in response to treatment with BR-WK. In addition, BR-WK reduced the number of tartrate-resistant acid phosphatase-positive ($TRAP^+$) multinucleated cells from bone marrow macrophages. These results indicate that BR-WK stimulates bone formation through its positive action on osteoblast differentiation and function and furthermore, decreases osteoclast differentiation.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.25 no.2
• /
• pp.1-11
• /
• 2012

Objectives: This study was conducted to evaluate the inhibitory effect of Melia Fructus extract on osteoclast differentiation. Methods: MTT-assay was performed to estimate cytotoxicity of Melia Fructus extract in BMMs stimulated with M-CSF. TRAP staining, TRAP activity and Real-time PCR were performed to know the inhibitory effect on osteoclast differentiation. Actin ring formation were analysed to observe the effect of Melia Fructus extract. Results: Melia Fructus extract decreased the number of TRAP positive cells and the expression of NFATc1 gene, c-Fos gene, TRAP and OSCAR in BMMs stimulated with RANKL. Melia Fructus extract has no cytotoxicity at the concentration used in this study. Melia Fructus extract restrained the formation of actin ring. Melia Fructus inhibited NF-${\kappa}B$ activity by inducing degradation of p-$IkB{\alpha}$. Conclusions: Melia Fructus has the inhibitory effect of osteocalst differentiation and bone resorption. Further studies are needed to treat osteoporosis by herbal medicine containing Melia Fructus.

• YAKHAK HOEJI
• /
• v.48 no.3
• /
• pp.197-201
• /
• 2004

Phosphodiesterases (PDEs) are enzymes that degrade intracellular cAMP. In the present study, pentoxifylline, a PDE inhibitor, induced osteoclast formation in co-cultures of mouse bone marrow cells and calvarial osteoblasts. To address the involvement of the osteoclast differentiation factor TNF-related activation-induced cytokine (TRANCE, identical to RANKL, ODF, and OPGL), mouse bone marrow cells and calvarial osteoblasts were co-cultured with pentoxifylline in the presence of OPG, a decoy receptor for TRANCE. The osteoclastogenic effect of pentoxifylline was completely blocked by addition of OPG, suggesting that TRANCE is involved in the osteoclast formation induced by pentoxifylline, Northern blot analysis revealed that pentoxifylline significantly induced TRANCE mRNA expression in calvarial osteoblasts. These results suggests that pentoxifylline regulates TRANCE expression in osteoblasts, which in turn controls osteoclast formation.

• Journal of Society of Preventive Korean Medicine
• /
• v.14 no.3
• /
• pp.13-26
• /
• 2010

Objectives : This study was performed to evaluate the effect of Securinega suffructicosa Rehder (SS) on osteoclast differentiation and gene expression. Methods : The osteoclastogenesis and gene expression were determined in RANKL-stimulated RAW 264.7. The results were summarized as followes. Results : SS decreased the number of TRAP positive cell in RANKL-stimulated RAW264.7 cell. SS decreased the expression of RANK, TNF${\alpha}$, and IL-6 in RANKL-stimulated RAW264.7 cell. SS decreased the expression of iNOS and COX-2 in RANKL-stimulated RAW264.7 cell. SS decreased the expression of Cathepsin K in RANKL-stimulated RAW264.7 cell. Conclusions : It is concluded that SS might decrease the bone resorption resulted from decrease of osteoclast differentiation and it's related gene expression.

• Herbal Formula Science
• /
• v.19 no.2
• /
• pp.61-71
• /
• 2011

Objectives : Kyungok-go(KOG), the first herbal formulation of donguibogam, has been used for treating of many symptoms of yin deficiency. In this study, we examined the effect of KOG on bone resorption. Methods : We determined the effects of water extract of KOG in RANKL(Receptor Activator for Nuclear Factor ${\kappa}B$ Ligand)-induced osteoclast differentiation culture system and osteoblast proliferation. In addition, we determined the effects of water extract of ABR on LPS-induced bone-loss with mice. Results : Water extract of KOG showed proliferation effect on osteoblast without cytotoxicity and no effect on RANKL-treated osteoclast differentiation. KOG rescued bone erosion by LPS induction in vivo study. Conclusions : These results demonstrated that KOG can be a useful remedy for treating of bone-loss disease such as osteoporosis.