scholarly journals Evaluation of Dexamethasone-Induced Osteoporosis In Vivo Using Zebrafish Scales

2021 ◽  
Vol 14 (6) ◽  
pp. 536
Author(s):  
Siripat Chaichit ◽  
Takuto Sato ◽  
Huiqing Yu ◽  
Yu-ki Tanaka ◽  
Yasumitsu Ogra ◽  
...  
Keyword(s):  
Enzyme Assay ◽  
Cathepsin K ◽  
Bone Diseases ◽  
Assay Method ◽  
Control Group ◽  
Tartrate Resistant Acid Phosphatase ◽  
Activity Levels ◽  
Insight Into ◽  
K Activity

Glucocorticoid-induced osteoporosis (GIOP) is a major cause of secondary osteoporosis, and the pathogenic mechanisms of GIOP remain to be elucidated. Here, we show a rapid dexamethasone-induced osteoporosis animal model using zebrafish scales. Intraperitoneal injection of dexamethasone over a 5-day period suppressed the regeneration of scales. Furthermore, the circularity of the newly formed regenerated scales was also slightly reduced compared to that of the control group on day 5. The changes in bone-related enzymes, such as cathepsin K, tartrate-resistant acid phosphatase (TRAP) for bone resorption, and alkaline phosphatase (ALP) for bone formation, provide insight into the progression of bone diseases; therefore, we further developed a method to measure the activities of cathepsin K, TRAP, and ALP using zebrafish scales. We found that a lysis buffer with detergent at neutral pH under sonication efficiently helped extract these three enzymes with high activity levels. Interestingly, treatment with a dexamethasone injection produced considerably higher levels of cathepsin K activity and a lower Ca/P ratio than those in the control group, suggesting that dexamethasone increased osteoclast activity, with no significant changes in the activities of TRAP and ALP. Our GIOP model and enzyme assay method could help to design better treatments for GIOP.

Cells in regenerating deer antler cartilage provide a microenvironment that supports osteoclast differentiation

2001 ◽  
Vol 204 (3) ◽  
pp. 443-455
Author(s):  
C. Faucheux ◽  
S. Nesbitt ◽  
M. Horton ◽  
J. Price
Keyword(s):  
Type I Collagen ◽  
Mononuclear Cells ◽  
Osteoclast Differentiation ◽  
Cathepsin K ◽  
Collagen Matrix ◽  
Tartrate Resistant Acid Phosphatase ◽  
Type I ◽  
Deer Antler

Deer antlers are a rare example of mammalian epimorphic regeneration. Each year, the antlers re-grow by a modified endochondral ossification process that involves extensive remodelling of cartilage by osteoclasts. This study identified regenerating antler cartilage as a site of osteoclastogenesis in vivo. An in vitro model was then developed to study antler osteoclast differentiation. Cultured as a high-density micromass, cells from non-mineralised cartilage supported the differentiation of large numbers of osteoclast-like multinucleated cells (MNCs) in the absence of factors normally required for osteoclastogenesis. After 48 h of culture, tartrate-resistant acid phosphatase (TRAP)-positive mononuclear cells (osteoclast precursors) were visible, and by day 14 a large number of TRAP-positive MNCs had formed (783+/−200 per well, mean +/− s.e.m., N=4). Reverse transcriptase/polymerase chain reaction (RT-PCR) showed that receptor activator of NF κ B ligand (RANKL) and macrophage colony stimulating factor (M-CSF) mRNAs were expressed in micromass cultures. Antler MNCs have the phenotype of osteoclasts from mammalian bone; they expressed TRAP, vitronectin and calcitonin receptors and, when cultured on dentine, formed F-actin rings and large resorption pits. When cultured on glass, antler MNCs appeared to digest the matrix of the micromass and endocytose type I collagen. Matrix metalloproteinase-9 (MMP-9) may play a role in the resorption of this non-mineralised matrix since it is highly expressed in 100 % of MNCs. In contrast, cathepsin K, another enzyme expressed in osteoclasts from bone, is only highly expressed in resorbing MNCs cultured on dentine. This study identifies the deer antler as a valuable model that can be used to study the differentiation and function of osteoclasts in adult regenerating mineralised tissues.

scholarly journals Blockade of NF-κB and MAPK pathways by ulinastatin attenuates wear particle-stimulated osteoclast differentiation in vitro and in vivo

2016 ◽  
Vol 36 (5) ◽  
Author(s):  
Jiang-Ying Ru ◽  
Hai-Dong Xu ◽  
Dai Shi ◽  
Jun-Bo Pan ◽  
Xiao-Jin Pan ◽  
...  
Keyword(s):  
Cathepsin K ◽  
Treated Group ◽  
Receptor Activation ◽  
Raw264.7 Cells ◽  
Control Group ◽  
Nuclear Factor ◽  
Trabecular Thickness ◽  
Tnf Α

Ulinastatin, a urinary trypsin inhibitor (UTI), is widely used to clinically treat lipopolysaccharide (LPS)-related inflammatory disorders recently. Adherent pathogen-associated molecular patterns (PAMPs), of which LPS is the best-studied and classical endotoxin produced by Gram-negative bacteria, act to increase the biological activity of osteopedic wear particles such as polymethyl-methacrylate (PMMA) and titanium particles in cell culture and animal models of implant loosening. The present study was designed to explore the inhibitory effect of UTI on osteoclastogenesis and inflammatory osteolysis in LPS/PMMA-mediated Raw264.7 cells and murine osteolysis models, and investigate the potential mechanism. The in vitro study was divided into the control group, LPS-induced group, PMMA-stimulated group and UTI-pretreated group. UTI (500 or 5000 units/ml) pretreatment was followed by PMMA (0.5 mg/ml) with adherent LPS. The levels of inflammatory mediators including tumour necrosis factor-α (TNF-α), matrixmetallo-proteinases-9 (MMP-9) and interleukin-6 (IL-6), receptor activation of nuclear factor NF-κB (RANK), and cathepsin K were examined and the amounts of phosphorylated I-κB, MEK, JNK and p38 were measured. In vivo study, murine osteolysis models were divided into the control group, PMMA-induced group and UTI-treated group. UTI (500 or 5000 units/kg per day) was injected intraperitoneally followed by PMMA suspension with adherent LPS (2×108 particles/25 μl) in the UTI-treated group. The thickness of interfacial membrane and the number of infiltrated inflammatory cells around the implants were assessed, and bone mineral density (BMD), trabecular number (Tb.N.), trabecular thickness (Tb.Th.), trabecular separation (Tb.Sp.), relative bone volume over total volume (BV/TV) of distal femur around the implants were calculated. Our results showed that UTI pretreatment suppressed the secretion of proinflammatory cytokines including MMP-9, IL-6, TNF-α, RANK and cathepsin K through down-regulating the activity of nuclear factor kappa B (NF-κB) and MAPKs partly in LPS/PMMA-mediated Raw264.7 cells. Finally, UTI treatment decreased the inflammatory osteolysis reaction in PMMA-induced murine osteolysis models. In conclusion, these results confirm the anti-inflammatory potential of UTI in the prevention of particle disease.

The In Vivo Activity of Neutralizing B Cell-Activating Factor (BAFF) Antibody in the Treatment of Multiple Myeloma (MM).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 843-843
Author(s):  
Paola Neri ◽  
Shaji Kumar ◽  
Mariateresa Fulciniti ◽  
Sonia Vallet ◽  
Shweta Chhetri ◽  
...  
Keyword(s):  
B Cell ◽  
Cell Survival ◽  
Plasma Cells ◽  
Neutralizing Antibody ◽  
Serum Levels ◽  
Survival Advantage ◽  
Control Group ◽  
Tartrate Resistant Acid Phosphatase ◽  
Growth And Survival

Abstract BAFF is a member of the tumor necrosis factor (TNF) family and plays a role in B cell survival including MM cells. We have previously established a role for BAFF in localization and survival of MM cells in the bone marrow (BM) microenvironment. (Cancer Res2006; 66(13): 6675–82). Here we validate the role of BAFF in MM patients and demonstrate the in vivo activity of anti-BAFF antibody in a SCID-hu MM model. We first performed gene expression profiling (GEP) on CD 138+ plasma cells isolated from 90 MM patients and 11 healthy controls using the Affymetrix U133A arrays. GEP analysis demonstrated increased BCMA expression (p<0.0001, student T test) on newly diagnosed and relapsed MM versus normal plasma cells. Flow cytometry performed on MM patient cells demonstrated the presence of all 3 receptors on CD 138+ cells. ELISA assays confirmed increased plasma BAFF levels in 51 MM plasma (mean: 1049 pg/ml; range: 176–4252 pg/ml) compared to 11 normal donors (mean: 461pg/ml; range: 317–652pg/ml) [p<0.001]. To understand the functional significance that BAFF might play in the biology of MM, we studied the effects of recombinant BAFF (rh-BAFF) on MM cells directly and in the context of its BM microenvironment. Our data demonstrate that rh-BAFF confers a survival advantage to MM cells and protects them against dexamethasone-induced cytotoxicity. Importantly, anti-apoptotic proteins Bcl2 and XIAP were upregulated, as were growth and survival signals belonging to the AKT and MAPKinase pathways. Because of the survival advantage conferred by BAFF on MM cells we evaluated the use of a clinical grade-neutralizing antibody to BAFF. To evaluate the in vivo activity of anti-Baff in MM we used the SCID-hu model, where MM cells grow in the context of human BM microenvironment. A cohort of SCID-hu mice bearing INA-6 MM cells, were treated i.p. weekly with anti-BAFF neutralizing antibody (10 mg/kg, n=3) or control isotype (10 mg/kg, n=3), respectively, for four weeks. Serum levels of soluble human IL-6 receptor (shuIL-6R) released by MM cells into the murine serum were monitored as a measure of MM growth. At the end of treatment we observed a significant (p=0.048) reduction in shuIL-6R level. This translated into a survival advantage of 3.1 weeks in the anti-BAFF treated animals versus the control group (p= 0.02). We also evaluated in vivo effects of anti-BAFF on the bone compartment by radiographic analysis and tartrate-resistant acid phosphatase (TRAP) staining of human bone implants. Our results demonstrate a decrease in radiologically evident lytic lesions in the anti-BAFF treated animals. This was accompanied by a significant decrease in TRAP + osteoclasts in bone sections from treated mice when compared to control mice. Our data therefore suggests that anti-BAFF antibody may impact MM cell growth directly and/or via effects on the BM microenvironment by impacting bone resorption. Taken together, these data show a role for BAFF mediating MM cell survival and its in vivo anti-tumor activities provide a preclinical rational for its clinical evaluation in MM.

scholarly journals Anti-Osteoporosis Effects of the Eleutherococcus senticosus, Achyranthes japonica, and Atractylodes japonica Mixed Extract Fermented with Nuruk

Nutrients ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 3904
Author(s):  
So Young Eun ◽  
Yoon-Hee Cheon ◽  
Gyeong Do Park ◽  
Chong Hyuk Chung ◽  
Chang Hoon Lee ◽  
...  
Keyword(s):  
Bone Loss ◽  
Bone Diseases ◽  
Marker Genes ◽  
Tartrate Resistant Acid Phosphatase ◽  
Specific Marker ◽  
Eleutherococcus Senticosus ◽  
Achyranthes Japonica ◽  
Atractylodes Japonica

Vigeo is a mixture of fermented extracts of Eleutherococcus senticosus Maxim (ESM), Achyranthes japonica (Miq.) Nakai (AJN), and Atractylodes japonica Koidzumi (AJK) manufactured using the traditional Korean nuruk fermentation method. Although the bioactive effects of ESM, AJN, and AJK have already been reported, the pharmacological effects of Vigeo have not been proven. Therefore, in this study, we investigated whether Vigeo had inhivitory effects on lipopolysaccharide (LPS)-induced inflammatory bone loss in vivo and receptor activator of nuclear factor-B ligand (RANKL)-induced osteoclastogenesis and the related mechanism in vitro. Vigeo administration conferred effective protection against bone loss induced by excessive inflammatory response and activity of osteoclasts in LPS-induced inflammatory osteoporosis mouse model. In addition, Vigeo significantly suppressed the formation of tartrate-resistant acid phosphatase-positive osteoclasts induced by RANKL and inhibited F-actin formation and bone resorbing activity without any cytotoxicity. Moreover, Vigeo significantly inhibited RANKL-induced phosphorylation of p38, ERK, JNK, IκB, and AKT and degradation of IkB. Additionally, Vigeo strongly inhibited the mRNA and protein expression of c-FOS and NFATc1 and subsequently attenuated the expression of osteoclast specific marker genes induced by RANKL. We demonstrated for the first time the anti-osteoporosis effect of Vigeo, suggesting that it could be a potential therapeutic candidate for the treatment of osteoclast-mediated inflammatory bone diseases.

A Novel Anilinoquinazoline Derivative Inhibits Growth of High-Risk Myeloma Cells In Vivo and Regulates Differentiation of Osteoclasts

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4076-4076
Author(s):  
Yoshie Ozaki ◽  
Yuhei Naito ◽  
Hiroshi Yanagawa ◽  
Yuya Suzuki ◽  
Wenlin Du ◽  
...  
Keyword(s):  
High Risk ◽  
Cell Lines ◽  
High Performance ◽  
Mononuclear Cells ◽  
Chromosomal Abnormalities ◽  
Conflicts Of Interest ◽  
Bone Diseases ◽  
Bone Lesions ◽  
Tartrate Resistant Acid Phosphatase

Abstract Abstract 4076 Purpose: Despite recent advances in the use of newly developed drugs, high-risk multiple myeloma (MM) patients harboring del13q, t(4;14) or del17p revealed significantly shorter survival. To overcome the limitation, we have screened forty synthetic anilinoquinazoline (AQ) derivatives, and found a novel compound, Q15, which significantly inhibited the growth of MM cell lines with high-risk chromosomal abnormalities. The purpose of this study is to examine anti-tumor and anti-osteoclastogenic activities of Q15 and to clarify the possibility of development of new drug effective for high-risk MM and bone diseases. Methods and Results: Forty AQ derivatives were synthesized and screened for anti-proliferative effect on KMS34 cells. Q15 strongly inhibited growth of t(4;14)-positive KMS34 cells and induced apoptosis in much lower concentration (IC50=78nM) compared with gefitinib (IC50=2500nM), a representative AQ. Q15 also inhibited growth of other MM cell lines harboring high-risk chromosomal abnormalities. It was also found that Q15 did not inhibit intracellular tyrosine phosphorylation induced by EGF, FGF-2, HGF and IL-6, suggesting that Q15 showed anti-tumor activity in a different mechanism from that of gefitinib. In vivo anti-myeloma activity was evaluated by intraperitoneal injection of Q15 into KMS34-bearing lcr/SCID mice. Twenty mg/kg Q15 significantly delayed the tumor growth in these mice. Histopathological examinations revealed apoptosis of MM cells in Q15-treated mice. Growth of colony-forming cells was not suppressed by much higher concentrations (25μ M) of Q15 than IC50, suggesting low hematopoietic toxicity of Q15. In pharmacokinetic study using high-performance liquid chromatography (HPLC), the plasma concentration of Q15 in mice reached a maximum (Cmax=4.5μ M) at 1.5hr after injection, and its half life (T1/2) was 4.5hr. In addition, anti-osteoclastogenic activity was also examined by adding Q15 to M-CSF/RANK ligand-induced osteoclastogenic culture of bone marrow mononuclear cells from C57BL/6JJcl mice. The number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated osteoclasts was reduced in the presence of Q15. Conclusion: Q15, a novel AQ derivative, has anti-MM activity in vivo and is a potentially safe and effective drug for high-risk MM with bone lesions. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Inhibitory Effects of 2N1HIA (2-(3-(2-Fluoro-4-Methoxyphenyl)-6-Oxo-1(6H)-Pyridazinyl)-N-1H-Indol-5-Ylacetamide) on Osteoclast Differentiation via Suppressing Cathepsin K Expression

Molecules ◽  
2018 ◽  
Vol 23 (12) ◽  
pp. 3139 ◽  
Author(s):  
Sun-Hee Ahn ◽  
Zhihao Chen ◽  
Jinkyung Lee ◽  
Seok-Woo Lee ◽  
Sang Min ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Osteoclast Differentiation ◽  
Cell Formation ◽  
Cathepsin K ◽  
Bone Diseases ◽  
Specific Gene ◽  
Tartrate Resistant Acid Phosphatase ◽  
Potential Candidate ◽  
Dependent Manner ◽  
Gene Markers

Osteoclasts are large multinucleated cells which are induced by the regulation of the receptor activator of nuclear factor kappa-Β ligand (RANKL), which is important in bone resorption. Excessive osteoclast differentiation can cause pathologic bone loss and destruction. Numerous studies have targeted molecules inhibiting RANKL signaling or bone resorption activity. In this study, 11 compounds from commercial libraries were examined for their effect on RANKL-induced osteoclast differentiation. Of these compounds, only 2-(3-(2-fluoro-4-methoxyphenyl)-6-oxo-1(6H)-pyridazinyl)-N-1H-indol-5-ylacetamide (2N1HIA) caused a significant decrease in multinucleated tartrate-resistant acid phosphatase (TRAP)-positive cell formation in a dose-dependent manner, without inducing cytotoxicity. The 2N1HIA compound neither affected the expression of osteoclast-specific gene markers such as TRAF6, NFATc1, RANK, OC-STAMP, and DC-STAMP, nor the RANKL signaling pathways, including p38, ERK, JNK, and NF-κB. However, 2N1HIA exhibited a significant impact on the expression levels of CD47 and cathepsin K, the early fusion marker and critical protease for bone resorption, respectively. The activity of matrix metalloprotease-9 (MMP-9) decreased due to 2N1HIA treatment. Accordingly, bone resorption activity and actin ring formation decreased in the presence of 2N1HIA. Taken together, 2N1HIA acts as an inhibitor of osteoclast differentiation by attenuating bone resorption activity and may serve as a potential candidate in preventing and/or treating osteoporosis, or other bone diseases associated with excessive bone resorption.

scholarly journals Contribution of Interleukin-11 and Prostaglandin(s) in Lipopolysaccharide-Induced Bone Resorption In Vivo

2002 ◽  
Vol 70 (7) ◽  
pp. 3915-3922 ◽  
Author(s):  
Li Li ◽  
Alireza Khansari ◽  
Lior Shapira ◽  
Dana T. Graves ◽  
Salomon Amar
Keyword(s):  
Bone Resorption ◽  
Subcutaneous Tissue ◽  
Interleukin 1 ◽  
Control Group ◽  
Tartrate Resistant Acid Phosphatase ◽  
Type I ◽  
Low Doses ◽  
Wild Type ◽  
Calvarial Bone

ABSTRACT We previously demonstrated that interleukin-1 (IL-1) and tumor necrosis factor (TNF) activities only partially account for calvarial bone resorption induced by local application of lipopolysaccharide (LPS) in mice. The present study was undertaken to determine the role and relative contribution of IL-11 and prostaglandin(s) (PG[s]) in LPS-induced bone resorption in vivo. A one-time dose of LPS was injected into the subcutaneous tissue overlying calvaria of mice lacking IL-1 receptor type I (IL-1RI−/−), mice lacking TNF receptor p55 and IL-1RI (TNFRp55−/−-IL-1RI−/−), and wild-type mice. Mice were then treated with injections of anti-IL-11 monoclonal antibody (MAb), indomethacin, or phosphate-buffered saline (PBS) and sacrificed 5 days later. Histological sections stained for tartrate-resistant acid phosphatase (TRAP) were quantified by histomorphometric analysis. At low doses of LPS (100 μg/mouse), the percentages of bone surface covered by osteoclasts were found to be similar in three strains of mice. The increase was reduced by 37% with anti-IL-11 MAb and by 46% with indomethacin. At higher doses of LPS (500 μg/mouse), we found an eightfold increase in these percentages in wild-type mice and a fivefold increase in these percentages in IL-1RI−/− and TNFRp55−/−−IL-1RI−/− mice after normalizing with the value from the saline-PBS control group in the same strain of mice. The increase was reduced by 55 and 69% in wild-type mice and by 50 and 57% in IL-1RI−/− and TNFRp55−/−−IL-1RI−/− mice treated with anti-IL-11 MAb or indomethacin, respectively. Our findings suggest that in vivo, at low doses of LPS (100 μg/mouse), LPS-induced bone resorption is mediated by IL-11 and PGs, while at high doses of LPS (500 μg/mouse), it is mediated by IL-11, PGs, IL-1, and TNF signaling. IL-11 and PGs mediate LPS-induced bone resorption by enhancing osteoclastogenesis independently of the IL-1 or TNF signaling.

scholarly journals PSTP-3,5-Me Inhibits Osteoclast Differentiation and Bone Resorption

Molecules ◽  
2019 ◽  
Vol 24 (18) ◽  
pp. 3346 ◽  
Author(s):  
Eunjin Cho ◽  
Zhihao Chen ◽  
Jinkyung Lee ◽  
Sunwoo Lee ◽  
Tae-Hoon Lee
Keyword(s):  
Osteoclast Differentiation ◽  
Cathepsin K ◽  
Osteoporosis Treatment ◽  
Bone Diseases ◽  
Marker Genes ◽  
Tartrate Resistant Acid Phosphatase ◽  
Mouse Bone Marrow ◽  
Nuclear Factor ◽  
Activated T Cell ◽  
Macrophage Colony

Osteogenesis is an orchestrated process regulated by osteoclastogenesis and osteoblastogenesis. Excessive osteoclastogenesis causes bone diseases, such as osteoporosis. Although a few drugs are effective in osteoporosis treatment, these drugs lead to side effects, including cellulitis, flatulence, and hypocalcemia. In this study, we reported a 2-(N-Phenylmethylsulfonamido)-N-(2-(phenylthio)phenyl)propanamide (PSTP) compound, PSTP-3,5-Me, as a potential therapeutic agent for osteoporosis. Mouse bone marrow-derived macrophages (BMMs) were differentiated into osteoclasts by receptor activator of nuclear factor kappa B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) in the presence of PSTP-3,5-Me. PSTP-3,5-Me inhibited osteoclast differentiation by reduced tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts, and suppressed the expression of osteoclast marker genes, such as cathepsin K (Ctsk) and TRAP (Acp5). We investigated signaling pathways mediated by RANKL and its receptor, RANK, and found that PSTP-3,5-Me inhibits nucleus translocation of nuclear factor of activated T cell cytoplasmic-1 (NFATc1). Moreover, PSTP-3,5-Me inhibited F-actin ring formation and mineral resorption. Overall, our data suggests that PSTP-3,5-Me attenuates osteoclast differentiation by blocking the activation of NFATc1.

scholarly journals N-[2-(4-Acetyl-1-Piperazinyl)Phenyl]-2-(3-Methylphenoxy)Acetamide (NAPMA) Inhibits Osteoclast Differentiation and Protects against Ovariectomy-Induced Osteoporosis

Molecules ◽  
2020 ◽  
Vol 25 (20) ◽  
pp. 4855
Author(s):  
Jinkyung Lee ◽  
Sun-Hee Ahn ◽  
Zhihao Chen ◽  
Sohi Kang ◽  
Dong Kyu Choi ◽  
...  
Keyword(s):  
Bone Resorption ◽  
Bone Loss ◽  
Osteoclast Differentiation ◽  
Cathepsin K ◽  
Bone Diseases ◽  
Drug Candidate ◽  
Tartrate Resistant Acid Phosphatase ◽  
Dependent Manner ◽  
Osteoclast Activity ◽  
Protein Levels

Osteoclasts are large, multinucleated cells responsible for bone resorption and are induced in response to the regulatory activity of receptor activator of nuclear factor-kappa B ligand (RANKL). Excessive osteoclast activity causes pathological bone loss and destruction. Many studies have investigated molecules that specifically inhibit osteoclast activity by blocking RANKL signaling or bone resorption. In recent years, we screened compounds from commercial libraries to identify molecules capable of inhibiting RANKL-induced osteoclast differentiation. Consequently, we reported some compounds that are effective at attenuating osteoclast activity. In this study, we found that N-[2-(4-acetyl-1-piperazinyl)phenyl]-2-(3-methylphenoxy)acetamide (NAPMA) significantly inhibited the formation of multinucleated tartrate-resistant acid phosphatase (TRAP)-positive cells from bone marrow-derived macrophages in a dose-dependent manner, without cytotoxic effects. NAPMA downregulated the expression of osteoclast-specific markers, such as c-Fos, NFATc1, DC-STAMP, cathepsin K, and MMP-9, at the transcript and protein levels. Accordingly, bone resorption and actin ring formation were decreased in response to NAPMA treatment. Furthermore, we demonstrated the protective effect of NAPMA against ovariectomy-induced bone loss using micro-CT and histological analysis. Collectively, the results showed that NAPMA inhibited osteoclast differentiation and attenuated bone resorption. It is thus a potential drug candidate for the treatment of osteoporosis and other bone diseases associated with excessive bone resorption.

scholarly journals Inhibitory effects of ChondroT and its constituent herbs on RANKL-induced osteoclastogenesis

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Rui Hong Guo ◽  
Seon-Jong Kim ◽  
Chan-hun Choi ◽  
Chang-su Na ◽  
Bok Yun Kang ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Osteoclast Differentiation ◽  
Cathepsin K ◽  
Mapk Signaling ◽  
Bone Diseases ◽  
Ring Formation ◽  
Tartrate Resistant Acid Phosphatase ◽  
Actin Ring ◽  
Specific Proteins ◽  
Underlying Mechanisms

Abstract Background ChondroT is a complex herbal medicine consisting of water extracts of Ostericum koreanum (Maxim.) Kitag., Lonicera japonica Thunb., Angelica gigas Nakai, Clematis manshurica Rupr., and Phellodendron amurense Rupr. (6:4:4:4:3). Previous studies have reported that ChondroT possesses chondroprotective and anti-inflammatory, anti-osteoarthritic, and anti-hyperuricemic activities. The study is aim to demonstrate the effects of ChondroT and its five constituent herbs on receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and the underlying mechanisms. Methods Osteoclastogenesis was identified in bone marrow-derived macrophages (BMDMs) by tartrate-resistant acid phosphatase (TRAP) staining assay, actin ring formation assay and the bone resorption assay. For the molecular mechanisms, activation of RANKL-induced NF-κB and MAPK signaling pathways and the expression levels of osteoclast-specific proteins were investigated by Western blotting. Cell viability was assessed by MTT assay. Actin ring formation and NF-κB translocation were evaluated by immunostaining. Results ChondroT and each of its constituent herbs significantly suppressed osteoclast differentiation dose dependently, and decreased actin ring formation as well as bone-resorbing capacity. Mechanistically, ChondroT and its constituent herbs downregulated the expressional levels of osteoclast-specific proteins such as NFATc1, c-Fos, Cathepsin K, and matrix metalloproteinase 9 (MMP9) by suppressing NF-κB translocation to nucleus and MAPKs phosphorylation at different levels. Compared to its five constituent herbs, ChondroT exhibited the best inhibitory efficiency against osteoclastogenesis. Conclusions Taken together, ChondroT has anti-osteoclastogenesis properties by inhibiting NF-κB and MAPKs pathways. It could be considered as a potential therapeutic candidate for the treatment of osteoclast-related bone diseases.

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