ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
Subject(s)
Animals , Humans , Male , Rats , Acid Phosphatase , Apoptosis , Caspase 3 , Caspase 8 , Caspases , Dental Cementum , Eosine Yellowish-(YS) , Hematoxylin , Immunohistochemistry , Models, Animal , Molar , Osteoclasts , Rats, Wistar , Root Resorption , Tooth Movement TechniquesABSTRACT
Objective To determine the protective effect of osteoprotegerin (OPG), receptor activator of nuclear factor kappa-B (RANK) and RANK ligand (RANKL) on avascular necrosis femoral head.Methods Necrotic tissue or corresponding normal tissues were collected from 29 avascular necrosis femoral head patients.Quantitative Real TimePCR ( qPCR ) is used to evaluate mRNA expression of OPG , RANK and RANKL.OPG and RANKL protein levels were estimated by Western blot.Results The results of qPCR showed that the expression of OPG in the necrotic tissue was significantly higher than that in the normal tissue (4.56 ±0.37) (3.39 ±0.52) (P<0.05).The expression levels of RANKL mRNA in necrotic tissues and normal tissues were (0.86 ±0.11) and (0.31 ±0.08), respectively,the difference was statistically significant (P<0.05).The expression levels of RANK mRNA in necrotic tissues and normal tissmes were(0.87 ±0.12), (0.56 ±0.13) respectively,the difference was statistically significant (P<0.05).The ratio of RANKL/OPG and RANK/OPG in normal tissues were 0.69 and 0.52, respectively.In the necrotic tissues, RANKL/OPG and RANK/OPG ratios were 1.35 and 0.61, respectively.Results of Western blot showed that the expression of OPG in necrotic tissues was consistent with that in normal tissues.The expression of RANKL protein was detected in all samples , and the expression of RANKL protein in necrotic tissue and normal tissue was almost the same.RANK protein expression was not detected in all samples.Conclusion OPG, RANK and RANKL play important roles in progress of bone remodeling in necrotic area and in disturbance of bone homeostasis and might have an effect on bone destruction and subsequent collapse of hip joint.
ABSTRACT
ObjectiveTo detect IL-17 level of bone marrow in patients with multiple myeloma bone disease,and to investigate its clinical significance.MethodsThe bone marrow IL-17 levels were quantified in 33 cases of multiple myeloma patients and 20 normal control by ELISA assay. RANKL mRNA expression were detected by using RT-PCR.ResultsIn bone marrow supernatant,IL-17 was detected in both groups,and RANKL mRNA were detected in both groups too. IL-17 and RANKL mRNA levels in bone marrow of patients with MM were significantly higher than those in the control group(P＜0.05). The bone marrow concentrations of IL-17 and bone marrow mononuclear cells' RANKL mRNA expression in active stage were significantly higher than those in stable stage (P＜0.05).The bone marrow IL-17 and RANKL were significantly correlated (r =690,P＜0.05).ConclusionIL-17 may play an important role in the pathogenesis of MM.
ABSTRACT
PURPOSE: Despite the fact that intraarticular corticosteroids are a longstanding adjuvant treatment for inflammatory arthritis, their mechanisms of action are not completely understood. This study evaluated the osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) levels in the serum and synovial fluid before and 2 weeks after injecting intraarticular corticosteroid. MATERIALS AND METHODS: Sixty seven patients were identified with joint effusions of the knee and suffering from rheumatoid arthritis. All the patients received an intraarticular injection of 40 mg of triamcinolone acetonide. The sera and synovial fluid were obtained before and 2 weeks after the injections. the erythrocyte sedimentation rate, C-reactive protein, rheumatoid factor, serum OPG and RANKL levels and synovial OPG and RANKL levels were measured before and 2 weeks after the injections. RESULTS: The intraarticular corticosteroids induced a significant decrease in the levels of synovial RANKL. The decrease in the level of synovial RANKL caused a significant increase in the synovial OPG/RANKL ratio. There were no changes in the hematological factors, serum OPG, serum RANKL or synovial OPG observed after the corticosteroid injection. CONCLUSION: Treatment with intraarticular corticosteroids modulates the OPG/RANKL system in the synovial fluid toward a bone-protective effect in patients with rheumatoid arthritis. This mechanism might explain part of the mode of action of intraarticular corticosteroids.
Subject(s)
Humans , Adrenal Cortex Hormones , Arthritis , Arthritis, Rheumatoid , Blood Sedimentation , C-Reactive Protein , Injections, Intra-Articular , Joints , Knee , Osteoprotegerin , RANK Ligand , Rheumatoid Factor , Stress, Psychological , Synovial Fluid , Triamcinolone AcetonideABSTRACT
PURPOSE: To clarify whether the bone resorption in femur measured by the expression of OPG & RANK-L was increased in tail suspended rat. MATERIALS AND METHODS: Four-week-old female Sprague Dawley rats were divided into two groups. The experimental group (n=79) was housed and fed with 2 weeks of tail suspension, and reloaded for 8 weeks without tail suspension. The control group (n=46) was housed and fed for 10 weeks without tail suspension. Bone mineral densities, serum levels of ALP and TRAP were measured in both groups. The expressions of the mRNAs of OPG and RANK-L were analyzed by RT-PCR. RESULTS: The ALP and TRAP were increased in the experimental group during both tail suspension and reloading, which reflected the increased bone metabolism in the experimental group. In femur of the experimental group, the expression of the mRNA of RANK-L was increased during tail suspension, and the expression the mRNA of OPG was decreased. With reloading, the expression of the mRNA of RANK-L in femur was decreased, while the expression of the mRNA of OPG was increased.
Subject(s)
Animals , Female , Humans , Rats , Bone Density , Bone Resorption , Femur , Hindlimb Suspension , Metabolism , Osteoclasts , Osteoporosis , Osteoprotegerin , Rats, Sprague-Dawley , RNA, Messenger , TailABSTRACT
Objective:To observe the effect of calcitonin gene-related peptide (CGRP) on the expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) in osteoblast cells through an in vitro breast cancer cell and osteoblast cell co-culture system. Methods:The metastatic breast cancer MDA-MB-231 or MDA-MB-435 cells were co-cultured with osteoblast MG63 cells to establish an in vitro microenvironment of bone metastasis of breast cancer. After treated with CGRP(1?108 mol/L),OPG and RANKL mRNA and protein expressions in osteoblast MG63 cells were examined by RT-PCR and Western blotting. Results:Expression of RANKL in osteoblast MG63 cells was up-regulated at both mRNA and protein levels when osteoblast MG63 cells were co-cultured with breast cancer MDA-MB-231 or MDA-MB-435 cells,while those of OPG in osteoblast MG63 cells were both down-regulated (P