• Korean Journal of Acupuncture
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• v.37 no.2
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• pp.63-75
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• 2020

Objectives : Osteoporosis is the most common bone disease and osteoporosis fracture is the leading cause of decreased life. Bisphosphonate and selective estrogen receptor modulators are the best choice of treatment for osteoporosis. However, when used for a long time, they increase the probability of side effect such as osteonecrosis of the jaw. Thus, it is crucial to develop alternative medicine to treat osteoporosis. Gentianae Macrophyllae Radix, a herbal medicine, is mainly to treat rheumatoid arthritis. However, the effect of the water extract of Gentianae Macrophyllae Radix (w-GM) on osteoporosis has not been investigated. Thus, we examine whether w-GM can inhibit osteoclast differentiation and bone resorption on receptor activator of nuclear factor kappa-B (NF-κB) ligand (RANKL)-treated RAW 264.7 cells. In this study, RAW 264.7 cells were used as an osteoclast differentiation model by treating them with RANKL. Methods : RAW 264.7 cells were used to determine the effect of w-GM on osteoclast differentiation and bone resorption. The number of tartrate-resistant acid phosphatase (TRAP)-positive cells, TRAP activity and pit formation assay were examined. In addition, protein expressions were measured by western blot and mRNA expressions were analyzed by reverse transcription polymerase chain reaction. Results : Treatment with w-GM inhibited the number of TRAP-positive cells, TRAP activity and pit area. In addition, w-GM decreased protein expression such as mitogen-activated protein kinase, NF-κB, c-Fos and nuclear factor of activated T-cells cytoplasmic 1 (NFATc1). It also inhibited the mRNA levels such as c-Fos, NFATc1, TRAP, NF-κB, calcitonin receptor and cathepsin K in RANKL-treated RAW 264.7 cells. Conclusions : These results suggest that w-GM has inhibitory effects via osteoclast differentiation, thus it could be a new medication for osteoporosis.

• Journal of Acupuncture Research
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• v.32 no.3
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• pp.27-40
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• 2015

Objectives : This study was performed to evaluate the effects of water extract of Cervi Parvum Cornu(CPC), Aconiti Lateralis Radix Preparata(ALR), and Yongbu-tang(YBT) on suppression of the receptor activator of nuclear factor kappa-B ligand(RANKL)-induced osteoclast differentiation and bone resorption. Methods : The effects of CPC, ALR, YBT extracts on osteoclast differentiation were determined by culture of bone marrow macrophage(BMM). The mRNA expression levels of the nuclear factor of activated T-cells cytoplasmic 1(NFATc1), c-Fos and tartrate-resistant acid phosphatase(TRAP) in BMMs were analyzed by reverse transcriptase polymerase chain reaction(RT-PCR). Similarly, the protein expression levels of NFATc1, c-Fos, mitogen-activated protein kinase(MAPK)s and ${\beta}$-actin in cell lysates were measured by western blotting. In addition, effects of CPC, ALR and YBT extracts were determined by means of Lipopolysaccharide(LPS)-induced bone-loss with mice. Results : CPC, ALR and YBT extracts showed remarkable inhibition on RANKL-induced osteoclast differentiation without cytotoxicity. CPC and ALR extracts significantly reduced the protein expression level of NFATc1. YBT extract significantly reduced the mRNA expression levels of c-Fos, NFATc1 and the protein expression levels of c-Fos, NFATc1, AKT, p38, c-Jun N-terminal kinase(JNK). Further, YBT extract suppressed degradation of$ I-{\kappa}B$. And ALR extract significantly restored the bone erosion by LPS treatment in mice. Conclusions : YBT extract showed more remarkable inhibition on osteoclast differentiation than CPC and ALR extracts in vitro. ALR extract showed remarkable inhibition on bone resorption in vivo. Thus, YBT extract can be a useful treatment for bone-loss diseases such as osteoporosis.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.516-520
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• 2011

Osteoporosis is social problem around the world, because fracture of old age may lead to critical medical condition. Osteoclast is a main target for prevention and treatment of osteoporosis due to their responsibility for bone resorption. Saussureae Radix has been known that has gastro-protective, bronchodilatory effect and has a anti-biotic effect. Saussureae Radix has been widely used in Oriental medicine. However, the effect of extract of Saussureae Radix in osteoclast differentiation remains unknown. Thus, we examined the effect of Saussureae Radix in receptor activator of nuclear factor-${\kappa}$B ligand (RANKL)-induced osteoclast differentiation. From the results of our study, Here we found that Saussureae Radix significantly inhibited osteoclast differentiation induced by RANKL. Saussureae Radix suppressed the activation of NF${\kappa}$B in bone marrow macrophages (BMMs) treated with RANKL. Also, Saussureae Radix significantly inhibited the mRNA expression of c-Fos, tartrate-resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), nuclear factor of activated T cells (NFAT)c1 and cathepsin K in BMMs treated with RANKL. Particularly, Saussureae Radix greatly inhibited the protein expression of c-fos and NFATc1. especially in the case of NFATc1 expression, a master transcription factor of the differentiation of osteoclasts is very important step for osteoclastogenesis. These results demonstrate that Saussureae Radix may be useful treatment option of bone-related disease such as osteoporosis and rheumatoid arthritis.

• The Journal of Internal Korean Medicine
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• v.43 no.6
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• pp.1134-1148
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• 2022

Purpose: We evaluated whether Spatholobus suberectus extract (SSE) can be used as a means of preventing and treating osteoporosis by measuring its effect on osteoclast differentiation, gene expression, and bone resorption. Methods: SSE was used to examine the effect on RAW 264.7 cells stimulated with RANKL to induce bone resorption. The inhibitory effect of TRAP formation and the expression of the bone resorption factors TRAP, cathepsin K, and MMP-9 during differentiation were measured. The effects on the differentiation-related factors NFATc and TRAIL and on the expression of OC-STAMP, DC-STAMP, ATP6v0d2, MITF, c-Fos, and inflammation-related factors were also evaluated. The effect on bone resorption was evaluated by culturing RANKL-treated osteoclasts on artificial bone fragments and observing the resulting resorption traces. The effect on bone damage in experimental animals was also measured. Results: SSE inhibited the differentiation of RANKL-stimulated osteoclasts into osteoclasts and suppressed the expression of cathepsin K, TRAP, MMP-9, NFATc1, TRAIL, MITF, OC-STAMP, DC-STAMP, ATP6v0d2, and c-Fos genes. Bone pore formation due to osteoclast action was also inhibited, and LPS-induced bone loss was suppressed in animal experiments. Conclusions: SSE could be useful for the prevention or treatment of osteoporosis by inhibiting osteoclast differentiation and bone resorption and suppressing bone loss induced in experimental animals. However, studies of larger populations are required.

• Biomolecules & Therapeutics
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• v.19 no.2
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• pp.141-148
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• 2011

Osteoclasts are bone-resorbing cells of monocyte/macrophage origin and are culprits of bone destruction associated with osteoporosis, rheumatoid arthritis, and cancer bone metastasis. Recent advances in osteoclast biology revealed central roles of various cytokines in regulating osteoclastogenesis both in vitro and in vivo. However, exact underlying mechanisms including signaling pathways downstream of receptor ligation are still under pursuit. In the present review, the role of Jak/STAT proteins and their regulators will be discussed in connection with osteoclastogenesis, since growing evidence indicates that a number of cytokines and growth factors utilizing Jak/STAT signaling pathways affect osteoclastogenesis. A better understanding on the role of Jak/STAT pathways in osteoclast differentiation will not only strengthen our knowledge on osteoclast biology but also provide invaluable insights into the development of anti-resorptive strategies for treating bone-lytic diseases.

• Korean Journal of Pharmacognosy
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• v.40 no.2
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• pp.83-88
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• 2009

Bone is continuously remodeled by osteoblasts and osteoclasts. Osteoclasts play an important role in bone metabolism by resorbing the bone matrix. Thus, the compounds inhibiting osteoclasts can improve bone diseases such as osteoporosis. The methanol extracts of 159 herbal medicines were screened for the inhibitory activity against differentiation of osteoclasts. Among the tested extracts, Achuranthis Radix and Corydalis Tuber showed relatively strong inhibitory activity against differentiation of osteoclasts, whereas they have no significant effect on proliferation of osteoclasts.

• Archives of Pharmacal Research
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• v.28 no.8
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• pp.909-913
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• 2005

We screened natural products to find compounds with anti-osteoporotic potential using a coculture-based system by which osteoclast differentiation is effectively achieved. We found that methylene chloride soluble fraction of the root of Salvia miltiorrhiza Bunge (Labiatae) suppressed osteoclast differentiation. Five tanshinones, tanshinone IIA (1), tanshinone I (2), cryptotanshinone (3), 15,16-dihydrotanshinone I (4), and ferruginol (5) were subsequently isolated from fraction. Among the five compounds, compounds 1-4 reduced the formation of TRAP­positive multinuclear osteoclasts. These results suggest that the identified tanshinones may be useful candidates for development of therapeutic agents to treat osteoporosis and other bone-resorptive diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.3
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• pp.320-324
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• 2012

Osteoporosis is an important public health issue in postmenopausal women. It is a major public health concern and is widely believed that osteoporosis results from imbalance between bone resorption and bone formation. Recently natural products from plants have been extensively studied as therapeutic drugs to treat and prevent various diseases. Hoelen (scientific name, Poria cocos) is a mushroom that is used in traditional Chinese medicine. Hoelen exhibits anti-inflammatory activity and has a protective effect on tumor progression. However, the effect of hoelen in osteoclast differentiation remains unknown. Thus, we examined the effect of hoelen in receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation. Hoelen significantly inhibited RANKL-induced osteoclast differentiation in bone marrow-derived macrophages (BMMs) in dose dependent manner without toxicity. Also, we showed that hoelen significantly inhibited the mRNA expression of tartrate-resistant acid phophatase (TRAP) and nuclear factor of activated T cells 1 (NFATc1) in BMMs treated with RANKL. In Particular, hoelen greatly inhibited the protein expression of NFATc1. Ectopic expression of NFATc1 partially reverses hoelen-mediated inhibition of osteoclast differentiation. Taken together, our results demonstrated that hoelen may be useful treatment option of bone-related disease such as osteoporosis, reumatoid arthritis, and periodontitis.

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

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