Effect of enhanced masticatory force on OPG, RANKL and MGF in alveolar bone of ovariectomized rats

Abstract Menopause induces oral bone loss, leading to various oral diseases. Mastication importantly affects bone metabolism in the jawbone. Objective: To analyze the effect of enhanced masticatory force on osteoprotegerin (OPG), receptor activator of nuclear factor kappa B ligand (RANKL), and mechano-growth factor (MGF) in alveolar bone of ovariectomized rats and to study the mechanics mechanism of the alveolar bone of ovariectomized rats response to enhanced masticatory force. Methodology: Thirty Sprague Dawley rats were randomly divided into three groups: sham-operation group (fat around the removed ovary + normal hard diet), model group (ovariectomy + normal hard diet), and experimental group (ovariectomy + high hard diet). It was a 2-month experiment. Enzyme-linked immunosorbent assay (ELISA) detected serum estradiol (E2), osteocalcin (BGP) and alkaline phosphatase (ALP) in rats. Bone histomorphometric indices in the third molar region of maxilla were detected by micro-CT; protein expressions of OPG, RANKL, and MGF in the third molar region of maxilla was detected by Western blot; and gene expression of OPG, RANKL, and MGF in the third molar region of maxilla was detected by Quantitative Real-Time PCR. Results: Comparing with model group, serum E2 in experimental group increased but not significantly, serum BGP and serum ALP in experimental group decreased but not significantly, OPG in experimental group in alveolar bone increased significantly, RANKL in experimental group in alveolar bone decreased significantly, RANKL/OPG ratio in experimental group decreased significantly, MGF in experimental group in alveolar bone increased significantly, bone volume to total volume fraction increased significantly in experimental group, trabecular thickness increased significantly in experimental group, and trabecular separation decreased significantly in experimental group. Conclusion: Enhanced masticatory force affected the expression of OPG, RANKL, and MGF in alveolar bone of ovariectomized rats, improved the quality of jaw bone of ovariectomized rats, and delayed oral bone loss by ovariectomy.

Effect of Decomposed Zuoguiwan Recipes in Improving BMD of Ovariectomized Rats by Regulating OPG/RANKL Pathway Mediated by β2AR / 中国实验方剂学杂志

Objective:To investigate the mechanism of decomposed Zuoguiwan（ZGW） recipes in treating ovariectomized osteoporosis rats. Method:Forty Sprague-Dawley female rats were equally and randomly divided into Sham-operated group, ovariectomized model group, positive group, and low and high-dose ZGW groups. After 12 weeks of administration by gavage, the bone mineral density (BMD) of rats' distal femur was measured by micro-CT, the morphology of bone tissue were observed by hematoxylin-eosin staining (HE), β-cross-linked c-telopeptide of type Ι collagen (β-CTX) and bone-specific alkaline phosphatase (BALP) in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the mRNA and protein expressions of β2AR, OPG and RANKL were evaluated by Western blot analysis and real-time quantitative polymerase chain reaction （PCR）. Result:Compared with Sham-operated group, BMD of rats in ovariectomized model group was decreased (P<0.01), morphology of bone tissue was destroyed, serum BALP was reduced, while β-CTX was boosted (P<0.01)，mRNA and protein expressions of OPG in tibia were reduced, while RANKL were increased, and mRNA and protein expressions of β2AR in the hypothalamus were decreased (P<0.05, P<0.01). Compared with ovariectomized model group, BMDs of rats in low and high-dose ZGW groups were increased (P<0.01), morphology of bone tissue was repaired, serum BALP and mRNA and protein expressions of OPG in tibia were up-regulated (P<0.05, P<0.01), whereas serum β-CTX and mRNA and protein expressions of β2AR in the hypothalamus and RANKL in tibia were down-regulated (P<0.05, P<0.01). Conclusion:Yang-nourishing components in decomposed Zuoguiwan recipes can improve BMD of ovariectomized rats by regulating OPG/RANKL pathway mediated by β2AR. "Seeking Yin in Yang" is a crucial mechanism of Zuoguiwan in treating ovariectomized osteoporosis in rats.

East java extract propolis as potential intracanal medicament in experimentally induced chronic apical periodontitis

Introduction: A persistent infection after cleaning and shaping root canal is the main etiology of root canal treatment failure. Enterococcus faecalis has been considered as one of the most resistant species in root canal treatment. E. faecalis can stimulate receptor activator of nuclear factor-kappa B ligand (RANKL) which can increase nuclear factor of activated T-cell (NFATc1) in chronic apical periodontitis. East Java propolis has antibacterial effects and is biocompatible with in vitro effects. Aim: This study is aimed to analyze the East Java propolis extract as potential intracanal medicament in chronic apical periodontitis caused by E. faecalis bacterial infection. Materials and Methods: This study used 30 Wistar rats divided into three groups. In Group I, the first upper right molar tooth as healthy tooth was used for negative control group. In Group II, the first upper right molar tooth was used for a prepared root canal, and 10 ml brain heart infusion broth containing E. faecalis ATCC29212 106 CFU was injected into the canal and restored with glass-ionomer cement (GIC) for the experimentally induced chronic apical periodontitis group. In Group III, after root canal preparation, E. faecalis ATCC 29212 106 CFU was injected, and then, 10 μl propolis applied and tooth restored with GIC. It took 21 days for the periapical lesions to develop after pulp infection. The rats were then sacrificed to conduct immunohistochemical examinations in order to measure the expressions of RANKL and NFATc1. Results: The average of RANKL and NFATc1 expression in Group III was significantly lower than those in the experimentally induced chronic apical periodontitis group (P < 0.05). Conclusion: It can be concluded that East Java propolis extract is a potential intracanal medicament through the study of experimentally induced chronic apical periodontitis caused by E. faecalis infection in Wistar rats.

Effect of Vitamin D during Orthodontic Tooth Movement on Receptor Activator of Nuclear Factor Kappa-Β Ligand Expression and Osteoclast Number in Pregnant Wistar Rat (Rattus novergicus)

Background: Female patients have the possibility tobecome pregnant during orthodontic treatment. VitaminD usually consumed by pregnant women. Estrogen andVitamin D could affect bone metabolism. Aim andObjectives: The aim of this study was to analyze theeffect of vitamin D during orthodontic movement inpregnant rats by Receptor Activator of Nuclear FactorKappa-Β Ligand (RANKL) expression and osteoclastnumber. Material and Methods: The experimentalobservational analytic study with post-test only controlgroup design and simple random sampling method wasconducted. 24-healthy-female Wistar rats were dividedinto 4 groups; K1: pregnant rats with orthodontic toothmovement and vitamin D on Day 7; K2: pregnant ratswith orthodontic tooth movement and vitamin D on Day14; K3: pregnant rats with orthodontic tooth movementwithout vitamin D on Day 7 and; K4: pregnant rats withorthodontic tooth movement without vitamin D on Day214. Nickle-Titanium coil spring with 10 g/mm forcewas placed between the incisors and the maxillarymolars. The RANKL expression and osteoclastsnumber were analyzed using Analysis of Variance(ANOVA) (p<0.05). Results: The highest osteoclastsnumber (8.494 ± 1.194), and RANKLexpression (7.967± 2.185) found in K1 group with significant betweengroups (p<0.05).Conclusions: Vitamin D increaseosteoclast number and RANKL expression duringorthodontic tooth movement in pregnant rats.

Effect of Zuoguiwan in Improving BMD and Trabecular Bone Microarchitecture in Ovariectomy-induced Osteoporosis Rats by Regulating RANKL/OPG Pathway Mediated by β2AR / 中国实验方剂学杂志

Objective: To investigate the mechanism of Zuoguiwan in treating ovariectomy-induced osteoporosis rats by receptor activator of nuclear factor kappa-B ligand (RANKL)/osteoprotegerin (OPG) signaling pathway mediated by β2-adrenergic receptor (β2AR). Method: Forty Sprague-Dawley female rats were randomly divided into Sham-operated group (Sham) and four ovariectomized (OVX) subgroups. Rats in Sham and OVX groups were treated with 17β-estradiol (50 μg·kg-1·d-1), and low and high-dose ZGW (2.3,4.6 g·kg-1 lyophilized powder) for 3 months, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to measure the serum markers of bone turnover. Micro-CT was used to evaluate and measure trabecular bone microarchitecture and bone mineral density (BMD) of the right distal femur. Western blot analysis and Real-time PCR were used to measure mRNA and protein expressions of β2AR, OPG and RANKL. Result: After 12 weeks of treatment with Zuoguiwan, the level of serum β-cross-linked c-telopeptide of type Ι collagen (β-CTX) (PPPβ2AR in the hypothalamus (PPConclusion: The mechanism of Zuoguiwan in alleviating BMD and trabecular bone microarchitecture in ovariectomy-induced osteoporosis rats might be related to the regulation of RANKL/OPG Pathway mediated by β2AR.

Efficacy of Wenyangbushen Formula on mRNA Expression of Osteoprotegerin, Receptor Activator of Nuclear Factor-kappa B and Its Ligand in Rabbits with Steroid-induced Avascular Necrosis of Femoral Head / 中国康复理论与实践

Objective:To observe the effect of Wenyangbushen formula on mRNA expression of osteoprotegerin (OPG), receptor activator of nuclear factor- kappa B (RANK) and receptor activator of nuclear factor- kappa B ligand (RANKL) in rabbits with steroid-induced avascular necrosis of femoral head (SANFH). Methods:A total of 46 healthy conventional New Zealand white rabbits were randomly divided into normal group (n = 10) and model building group (n = 36). The modified method of horse serum plus methylprednisolone was used to establish the SANFH model. Two rabbits from the normal group and four from the model building group were used for HE staining. Then the other models were randomly divided into model group, and low-dose, medium-dose and high-dose treatment groups, with eight rabbits in each group. The normal group was given normal saline 10 ml/d, and the treatment groups were given Wenyangbushen formula 6.44 g/(kg·d), 9.66 g/(kg·d) and 12.88 g/(kg·d), respectively, for eight weeks. The mRNA expression of OPG, RANK and RANKL was detected by reverse transcription- polymerase chain reaction. Results:The empty lacuna rate was significantly higher in the model group than in the normal group (t = 17.085, P < 0.001). Compared with the model group, the mRNA expression of OPG increased (P < 0.01), and the mRNA expression of RANK and RANKL decreased (P < 0.01), in the treatment groups. Compared with the low-dose treatment group, the mRNA expression of OPG increased (P < 0.01), and the mRNA expression of RANK and RANKL decreased (P < 0.01), in the medium-dose and high-dose treatment groups. There was no significant difference in the mRNA expression of OPG, RANK and RANKL between the low-dose treatment group and the high-dose treatment group (P > 0.05). Conclusion:Wenyangbushen formula could increase the mRNA expression of OPG and inhibit the mRNA expression of RANK and RANKL in the femoral head tissue of the rabbits with SANFH.

Effect of caspases and RANKL induced by heavy force in orthodontic root resorption / 대한치과교정학회지

OBJECTIVE: Orthodontic root resorption (ORR) due to orthodontic tooth movement is a difficult treatment-related adverse event. Caspases are important effector molecules for apoptosis. At present, little is known about the mechanisms underlying ORR and apoptosis in the cementum. The aim of the present in vivo study was to investigate the expression of tartrate-resistant acid phosphatase (TRAP), caspase 3, caspase 8, and receptor activator of nuclear factor kappa-B ligand (RANKL) in the cementum in response to a heavy or an optimum orthodontic force. METHODS: The maxillary molars of male Wistar rats were subjected to an orthodontic force of 10 g or 50 g using a closed coil spring. The rats were sacrificed each experimental period on days 1, 3, 5, and 7 after orthodontic force application. And the rats were subjected to histopathological and immunohistochemical analyses. RESULTS: On day 7 for the 50-g group, hematoxylin and eosin staining revealed numerous root resorption lacunae with odontoclasts on the root, while immunohistochemistry showed increased TRAP- and RANKL-positive cells. Caspase 3- and caspase 8-positive cells were increased on the cementum surfaces in the 50-g group on days 3 and 5. Moreover, the number of caspase 3- and caspase 8-positive cells and RANKL-positive cells was significantly higher in the 50-g group than in the 10-g group. CONCLUSIONS: In our rat model, ORR occurred after apoptosis was induced in the cementum by a heavy orthodontic force. These findings suggest that apoptosis of cementoblasts is involved in ORR.

Bone-targeted therapies to reduce skeletal morbidity in prostate cancer / 亚洲男科学杂志(英文版)

Bone metastases are the main driver of morbidity and mortality in advanced prostate cancer. Targeting the bone microenvironment, a key player in the pathogenesis of bone metastasis, has become one of the mainstays of therapy in men with advanced prostate cancer. This review will evaluate the data supporting the use of bone-targeted therapy, including (1) bisphosphonates such as zoledronic acid, which directly target osteoclasts, (2) denosumab, a receptor activator of nuclear factor-kappa B (RANK) ligand inhibitor, which targets a key component of bone stromal interaction, and (3) radium-223, an alpha-emitting calcium mimetic, which hones to the metabolically active areas of osteoblastic metastasis and induces double-strand breaks in the DNA. Denosumab has shown enhanced delay in skeletal-related events compared to zoledronic acid in patients with metastatic castration-resistant prostate cancer (mCRPC). Data are mixed with regard to pain control as a primary measure of efficacy. New data call into question dosing frequency, with quarterly dosing strategy potentially achieving similar effect compared to monthly dosing for zoledronic acid. In the case of radium-223, there are data for both pain palliation and improved overall survival in mCRPC. Further studies are needed to optimize timing and combination strategies for bone-targeted therapies. Ongoing studies will explore the impact of combining bone-targeted therapy with investigational therapeutic agents such as immunotherapy, for advanced prostate cancer. Future studies should strive to develop biomarkers of response, in order to improve efficacy and cost-effectiveness of these agents.

Expression of osteoprotegerin and receptor activator of nuclear factor kappa-B ligand in mandibular ramus osteotomy healing with administration of different doses of parathyroid hormone / 中华口腔医学杂志

Objective@#To investigate the effect of parathyroid hormone (PTH) on the bone healing of mandibular ramus osteotomy.@*Methods@#The mandibular ramus osteotomy model was established in sixty rabbits and these rabbits were randomly divided into experimental group A, experimental group B and control group. In the experimental group A and experimental group B, the rabbits were given PTH (20 and 40 μg/kg respectively) every other day after operation. In the control group, 1 ml saline was given. The animals were sacrificed at 1 week, 2 weeks, 3 weeks and 4 weeks postoperatively. The new bone formation was observed by histology and cone bone CT. The expression of osteoprotegerin and receptor activator of nuclear factor kappa-B (RANKL) in the new bone was detected by real-time quantitative PCR.@*Results@#The experimental groups has better osteogenesis and the bone mineral density than the control group in osteotomy area. The experimental group B showed the best osteogenesis.Osteoprotegerin mRNA expression in experimental group A (1.127±0.035, 1.742±0.049, 1.049±0.062, 1.063±0.036) was significantly higher than that in the control group in each period (0.965±0.082, 1.254±0.071, 0.793±0.061, 0.684±0.055) (P=0.010, P=0.000, P=0.001, P=0.020), while group B (1.416±0.205, 2.648±0.168, 1.652±0.091, 1.712±0.070) was significantly higher than group A (P=0.000, P=0.010, P=0.023, P=0.003). RANKL mRNA expression in control group (1.666±0.086, 1.058±0.105, 0.885±0.124, 0.972±0.136) was significantly higher than that of the group A (0.788±0.036, 0.585±0.017, 0.692±0.017, 0.527±0.051) (P=0.001, P=0.006, P=0.003, P=0.028) in each period, while group A was significantly higher than group B(0.247±0.022, 0.240±0.034, 0.134±0.011, 0.103±0.050) (P=0.000, P=0.001, P=0.002, P=0.012).@*Conclusions@#PTH can upregulate the expression of osteoprotegerin and reduce expression of RANKL, thus promoting new bone formation. Intermittent administration of high dose of parathyroid hormone can further promote the healing process after mandibular ramus osteotomy.

The effects of different concentrations of crocin on receptor activator of nuclear factor kappa B ligand(RANKL)–induced osteoclastogenesis / 天津医药

Objective To investigate the effects of different concentrations of crocin on receptor activator of nuclear factor kappa B ligand (RANKL)-induced osteoclastogenesis using the monocyte-macrophage cell line RAW264.7. Methods The monocyte-macrophage cell line RAW264.7 was cultured routinely.After treatment with 0,6.25,12.5,25, 50,100,200 and 400 μmol/L crocin,cell counting kit-8(CCK-8)assay was used to analyze the viability of RAW264.7 cells to screen out the appropriate experimental concentration. RAW264.7 cells were induced by RANKL (100 ng/L) to form osteoclasts. After treated with 0, 12.5, 50 and 100 μmol/L crocin respectively, the number of osteolasts was counted by tatrate resistant acid phosphatasec (TRAP) staining. Real-time PCR was used to detect the mRNA expression levels of calcitonin receptor(CTR),nuclear factor of active T cells 1(NFATC1),C-fos and TRAP.Results No significant effects of crocin (within 0-100 μmol/L) were found on the viability of RAW264.7 cells (P>0.05). However, When crocin concentration was over 100 μmol/L,the cell proliferation was decreased,and which showed a significant inhibitory effect on proliferation (P<0.05). Thus, 0-100 μmol/L crocin was selected as the experiment concentration. The amount of differentiated osteolasts and the expression levels of CTR,NFATC1,C-fos and TRAP mRNA were decreased significantly with the increased concentrations of crocin(P<0.05).Conclusion At a certain concentration(0-100 μmol/L),the higher levels of crocin could inhibit RANKL-induced osteoclastogenesis.

Effect of Wenhua Juanbi Recipe () on expression of receptor activator of nuclear factor kappa B ligand, osteoprotegerin, and tumor necrosis factor receptor superfamily member 14 in rats with collagen-induced arthritis / 中国结合医学杂志

<p><b>OBJECTIVES</b>To study the effect of Wenhua Juanbi Recipe (, WJR) on expression of receptor activator of nuclear factor kappa B ligand (RANKL), osteoprotegerin (OPG), and tumor necrosis factor receptor superfamily member 14 (TNFRSF14, also known as LIGHT) in rats with collagen-induced arthritis (CIA).</p><p><b>METHODS</b>CIA rats were generated by subcutaneous injection of bovine collagen type-II at the tail base. Sixty CIA rats were randomly assigned (10 animals/group) to: model, methotrexate (MTX)-treated (0.78 mg/kg body weight), and WJR-treated (22.9 g/kg) groups. Healthy normal rats (n=10) were used as the normal control. Treatments or saline were administered once daily by oral gavage. Rats were sacrifificed at day 28 post-treatment and knee synovium and peripheral blood serum were collected. Toe swelling degree and expression of RANKL, OPG, and LIGHT were determined by Western blot and immunohistochemistry.</p><p><b>RESULTS</b>Compared with the normal group, toe swelling degree was signifificantly increased in the model group (P<0.01). After treatment, toe swelling degree decreased signifificantly in the WJR and MTX groups compared with the model group (P<0.01). Compared with the normal group, expression of RANKL and LIGHT were signifificantly increased and OPG signifificantly decreased in peripheral blood and synovium of the model group (P<0.01). Conversely, RANKL and LIGHT expression were signifificantly reduced and OPG increased in the WJR and MTX groups compared with the model group (P<0.01). No statistically significant difference existed between WJR and MTX groups.</p><p><b>CONCLUSION</b>WJR likely acts by reducing RANKL expression and increasing OPG expression, thus inhibiting RANKL/RANK interaction and reducing LIGHT expression, thereby inhibiting osteoclast formation/activation to block bone erosion.</p>

Effect of sumac extract on serum oxidative status, RANKL/OPG system and alveolar bone loss in experimental periodontitis in rats

Objectives Sumac (Rhus coriaria L.) is widely used spice which has several properties such as antioxidant, anti-inflammatory and antimicrobial. The purpose of this animal study was to evaluate the effects of sumac extract on levels of receptor activator of nuclear factor-kappa B ligand (RANKL), osteoprotegerin (OPG) expression, serum oxidative status, and alveolar bone loss in experimental periodontitis. Material and Methods Twenty-four Wistar rats were separated into three groups: non-ligated (NL, n=8), ligature only (LO, n=8), and ligature and treated with sumac extract (S, n=8) (20 mg/kg per day for 11 days). A 4/0 silk suture was placed around the mandibular right first molars subgingivally; after 11 days, the rats were sacrificed, and alveolar bone loss was histometrically measured. The detection of RANKL and OPG were immunohistochemically performed. Levels of serum total antioxidant status (TAS)/total oxidant status (TOS), and oxidative stress index (OSI) were also analyzed. Results Alveolar bone loss was significantly greater in the LO group compared to the S and NL groups (p<0.05). The number of inflammatory cell infiltrate (ICI) and osteoclasts in the LO group was significantly higher than that of the NL and S groups (p<0.05). The number of osteoblasts in the LO and S groups was significantly higher than that of the NL group (p<0.05). There were significantly more RANKL-positive cells in the LO group than in the S and NL groups (p<0.05). OPG-positive cells were higher in S group than in LO and NL groups (p<0.05). TOS and OSI levels were significantly reduced in S group compared to LO group (P<0.05) and TAS levels were similar in S and NL group (p>0.05). Conclusions The present study showed that systemic administration of sumac extract may reduce alveolar bone loss by affecting RANKL/OPG balance, TOS and OSI levels in periodontal disease in rats. .

The Time Course Changes in Bone Metabolic Markers after Administering the Anti-Receptor Activator of Nuclear Factor-Kappa B Ligand Antibody and Drug Compliance among Patients with Osteoporosis

STUDY DESIGN: Retrospective study. PURPOSE: We conducted a study to investigate the time course changes in bone metabolic markers after the administration of the anti-receptor activator of nuclear factor-kappa B ligand (RANKL) antibody and to assess drug compliance among osteoporotic patients. OVERVIEW OF LITERATURE: The anti-RANKL antibody is expected to provide an improvement in those with a bone metabolism disorder. However there are only a few clinical reports available on the effect of treatment. METHODS: We included 40 post-menopausal osteoporotic patients who received the anti-RANKL antibody. To determine the time course changes in the bone metabolic markers, we measured the serum tartrate-resistant acid phosphatase 5b (TRACP 5b; a bone resorption marker) and the serum N-terminal propeptide of type 1 collagen (P1NP; a bone formation marker) levels prior to and 1 month after administrating the anti-RANKL antibody. To evaluable drug compliance, we assessed the dropout rate during treatment and at 6 months after treatment. RESULTS: The average TRACP 5b level significantly decreased from 574.8 mU/dL before treatment to 153.2 mU/dL 1 month after treatment (p0.05). As for drug compliance, we did not have any dropouts during the treatment or after 6 months (dropout rate: 0%). CONCLUSIONS: Our study suggests that anti-RANKL antibody treatment suppresses bone resorption and maintains bone formation.

Effects of parthenolide on osteoclast differentiation of RAW264. 7 cell induced by receptor activator of ;nuclear factor κB ligand / 中华风湿病学杂志

Objective To study the effects of parthenolide on osteoclast differentiation of RAW264. 7 cell induced by receptor activator of nuclear factor κB ligand (RANKL). Methods The mouse macrophage RAW264.7 cells induced by RANKL was used alone as the control group, different concentrations of par-thenolide (0.5, 1, 2 μmol/L) were added to culture the RAW264.7 cells. On the third, fifth and seventh day, the tartrate resistant acid phosphatase (TRAP) staining method was used to detect osteodast-like cells and the cell number was count;the contents of tartrate resistant acid phosphatase (TRAP5b) in the Culture supernatant of each groups were detected by enzyme linked immunosorbent assay (ELISA) and the expression of osteodast marker gene alcitonin receptor (CTR) and matrix metalloproteinase (MMP)-9 in each groups were detected by realtime-polymerase chain reaction (PCR) on the seventh day. We use Chi-square test and t test to test the differences between groups by SPSS 17.0. Results In different culture conditions, RANKL could always induce the RAW264.7 cell differentiate into mature osteoclasts. Compared with the control group at the same time control group, on the third, fifth and seventh day, he number of mature osteoclasts induced were obviously decreased in groups adding different concentration of PAR; the number of induced osteoclasts decreased along with the increase of parthenolide concentration, on the seventh day in 0.5, 1, 2 μmol/L concentration PAR groups, the number of mature osteoclasts compared with the control group were descended 36.3%, 40.8%, 49.3%(t=7.758, 8.742, 10.56;P<0.05);the contents of TRAP5b in the culture supernatant were consistent with the cell counting results on the seventh day (P<0.05). The expression of CTR and MMP-9 by TRAP positive osteoclasts decreased along with the increase of parthenolide concentration, and the 2 μmol/L group was the lowest. Compared with the control group, there were statistically significant differences with the different PAR concentration groups 0.5, 1, 2 μmol/L (P<0.05). Conclusion Parthenolide can inhibit RANKL induced RAW264.7 differentiation into osteoclast cells, and the inhibition is dose dependent.

Effect of curcumin on expression of synovial osteoprotegerin and receptor activator of nuclear factor-κB ligand protein in adjuvant arthritis rats / 中国免疫学杂志

Objective:By observing the effects of curcumin on synovial pathology, synovial osteoprotegerin and receptor activator of nuclear factor κB ligand expression factor of adjuvant arthritis rat models, we try to explore the possible mechanism of prevention and treatment of rheumatoid arthritis.Methods: There were three groups, the normal group, the model group and the curcumin group.We made adjuvant arthritis rat models with Freund′s complete adjuvant,and they were divided into the model group and the curcumin group.On d 28 after modeling,all rats were killed.All synovial membranes were sent for routine histological evaluation with HE staining.And synovial RANKL and OPG protein expression were measured by Western blot.Results: Curcumin could influence synovium pathology change of adjuvant arthritis rats,the pathological picture showed that infiltration of joint synovial cell,pro-liferation of synovial fibroblasts in model group rats were significantly increased,compared with normal group( P<0.01).Pathology of rats synovial tissue in rats of curcumin group were significantly improved, compared with that in rats of model group.Expression of RANKL,OPG protein in synovial tissue of curcumin group,based on pairwise comparison between with model group,the results were statistically significant( P<0.01 ).Synovial RANKL/OPG ratio were significantly lower than model group ( P<0.01 ).Conclusion:Curcumin can improve synovial pathology damage of adjuvant arthritis rats,curcumin may exert a protective effect on arthritis rats by regulating OPG/RANKL signaling pathway.

Evaluation of Behavior and Expression of Receptor Activator of Nuclear Factor-Kappa B Ligand in Dorsal Root Ganglia after Sciatic Nerve Compression and Application of Nucleus Pulposus in Rats

STUDY DESIGN: Experimental animal study. PURPOSE: To evaluate pain-related behavior and changes in nuclear factor-kappa B (NF-kB), receptor activator of NF-kB (RANK), and ligand (RANKL) in dorsal root ganglia (DRG) after combined sciatic nerve compression and nucleus pulposus (NP) application in rats. OVERVIEW OF LITERATURE: The pathological mechanisms underlying pain from lumbar-disc herniation have not been fully elucidated. RANKL are transcriptional regulators of inflammatory cytokines. Our aim was to evaluate pain-related behavior and RANKL expression in DRG after sciatic-nerve compression and application of NP in rats. METHODS: Mechanical hyperalgesia and RANKL expression were assessed in three groups of rats: NP+sciatic nerve compression (2 seconds), sham-operated, and controls (n=20 each). Mechanical hyperalgesia was measured every other day for 3 weeks using von Frey filaments. RANKL expression in L5 DRGs was examined at five and ten days after surgery using immunohistochemistry. RESULTS: Mechanical hyperalgesia was observed over the 12-day observation period in the NP+nerve compression group, but not in the control and sham-operated animal groups (p<0.05). RANKL immunoreactivity was seen in the nuclei of L5 DRG neurons, and its expression was significantly upregulated in NP+nerve compression rats compared with control and sham-operated rats (p<0.01). CONCLUSIONS: The exposure of sciatic nerves to mechanical compression and NP produces pain-related behavior and up-regulation of RANKL in DRG neurons. RANKL may play an important role in mediating pain after sciatic nerve injury with exposure to NP.

Chinese medicine Gukang prescription modulates core binding factor alpha 1 expressing in osteoblasts / 中国组织工程研究

BACKGROUND: Chinese medicine Gukang prescription has a clear effect on clinical treatment of osteoporosis, but the therapeutic pathway is stil unclear. OBJECTIVE:To investigate the effects of Chinese medicine Gukang on the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin by regulating core binding factor alpha 1 expression to control the growth and development of osteoblasts. METHODS:Sprague-Dawley neonatal rats within 24 hours after delivery were used for the separation and culture of osteoblasts. Adult Sprague-Dawley rats were used to prepare drug-containing serum, and then divided into two groups randomly:normal control group and Gukang group. Rats in the normal control and Gukang groups were intragastrical y administrated with extract of Gukang prescription and normal saline based on rat’s body surface area, for 1 consecutive week. Two hours after the last administration, blood samples were taken from the heart. Then the serum was col ected. Osteoblasts at passage 3 were confirmed with alkaline phosphatase assay and digested. After counting and planting, al osteoblasts were divided into two groups and treated with col ected serum for 72 hours. Proliferative rate of osteoblasts was detected by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. Secretion of alkaline phosphatase was detected by using enzyme linked immunosorbent assay and corrected with the corresponding absorbance value. mRNA expression of core binding factor alpha 1, receptor activator of nuclear factor kappa B ligand and osteoprotegerin were detected by using reverse transcription-PCR in al groups. RESULTS AND CONCLUSION:mRNA expression of osteoprotegerin and core binding factor alpha 1 in the Gukang group was significantly higher than that in the normal control group, but protein and mRNA expression of receptor activator of nuclear factor kappa B ligand were dramatical y lower in the Gukang group compared with the normal control group (Pcore binding factor alpha 1, thereby adjusting the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin, which may be one of the mechanisms underlying Gukang treatment for osteoporosis.

The changes and clinical significance of osteoprotegerin and receptor activator of nuclear factor kappa B ligand in patients with chronic heart failure / 中国医师进修杂志

Objective To explore the plasma levels and clinical significance of osteoprotegerin(OPG)and receptor activator of nuclear factor kappa B ligand(RANKL)in patients with chronic heart failure(CHF).Methods In 110 patients with CHF and 80 normal controls,the plasma levels of OPG and RANKL were measured by enzyme-linked immunosorbentassay method.Results The plasma level of OPG in patients with CHF[(135.91±41.83)ng/L]was significantly higher than that in normal controls[(90.13±29.09)ng/L](P＜0.05).The plasma level of RANKL in patients with CHF[(90.82±32.14)ng/L]was significantly higher than that in normal controls[(59.78±20.11)ng/L](P＜0.05).The plasma levels of OPG and RANKL increased with the augment of NYHA functional ranking(P＜0.05).The plasma levels of OPG and RANKL had negative relationship with left ventricular ejection fraction(r=-0.33 and-0.36,P＜0.05).Conclusions The plasma levels of OPG and RANKL in patients with CHF are elevated and related with heart function.OPG and RANKL may participate in the occurrence and development of CHF.

The effects of osteoclasts and osteoprotegerin/receptor activator of nuclear factor-kappa B ligand system on bone destruction of psoriatic arthritis / 中华风湿病学杂志

ObjectiveTo explore the effect of osteoclasts and osteoprotegerin/receptor activator of nuclear factor-kappa B ligand (OPG/RANKL) system on bone destruction of psoriatic arthritis.MethodsThe peripheral blood mononuclear cells from 41 psoriatic arthritis (PsA) patients,20 osteoarthritis (OA) patients and 24 healthy controls were cultured to become osteoclasts.After 14 days,cytochemistry method was used to detect tartrate-resistant acid phosphatase (TRAP) expression.At the same time,enzyme-linked immuno sorbent assay(ELISA) was used to measure the levels of serum OPG and RNAKL for all cases.At the same time,the clinical and laboratory examinations of the PsA were collected.Statistical analysis was conducted with one-way ANOVA and Spearman's correlation.ResultsSignificantly higher OC production was observed in the peripheral blood of PsA patients[(17.7±4.8)/view field] than that of the healthy controls[(6.4±1.6)/view field] and OA patients [(6.5±l.6)/view field].The levels of RANKL were significantly higher in PsA patients [(178±38) pg/ml] than those in the other two groups [(32±4) pg/ml and (67±17) pg/ml].There was significant difference between the PsA group with bone destruction and without destruction in the levels of OC and RANKL [(17.6±0.9) /view vs(7.9±1.3) /view and(199±72) pg/ml vs(128±44) pg/ml,P＜0.01].Imaging scores was positively correlated with the levels of OC and RANKL in PsA patients (P＜0.05).ConclusionIn PsA,there are significantly more OC and higher RANKL in the peripheral blood than those of the controls.The high levels of OC and RANKL are closely related with bone destruction.OC and RANKL are useful in identifying bone destruction.

Osteolysis effect by fibroblasts derived from malignant bone tumor / 中国矫形外科杂志

[Objective]To investigate whether fibroblasts from malignant bone tumor could support osteoclast differentiation and bone resorption.[Method]Fibroblasts isolated from 8 fresh tumor samples were cultured in vitro.At passage two,part of these fibroblasts were co-cultured with peripheral blood monocytes.M-CSF,OPG and neutralizing anti-TNF-? antibody was added to the culture respectively.A Transwells system was also applied to observe the secretion of solvable factors in fibroblasts.At day 1,day 14 and day 21,the cultures on coverslips and bone slices were stopped and examined for the formation of tartrate resistant acid phosphatase(TRAP)positive multmucleated cells(MNCs)and lacuna resorption pits respectively.The number of lacuna resorption pits on bone slices,which was a marker of OC activity,was compared between the groups.Another part of the fibroblasts were cultured alone,and RT-PCR was performed to investigate the expression of RANKL and TNF-? mRNA in the fibroblasts.[Result]TRAP positive MNCs and lacuna pits were observed in the co-culture group in the presence of M-CSF,but not in the absence of M-CSE.It was also the case in the Transwell group,even though the number of lacuna pits was resuced.OPG completely inhibited the formation of TRAP positive MNCs and lacuna pits,while anti-TNF-? had not inhibiting effect on this process.RT-PCR results showed that both RANKL and TNF-? mRNA were positive in all the fibroblast involved.[Conclusion]In the presence of M-CSF,fibroblasts from malignant bone tumor are capable of supporting osteoclast formation and activity through expression of RANKL.