Journal: Scientific reports

Article Title: Black tea rich in polygonatum sibiricum polysaccharides inhibits osteoporosis in ovariectomized mice through immune regulation and gut microbiota suppression.

doi: 10.1038/s41598-025-08827-0

Figure Lengend Snippet: Fig. 1. (A-C) Changes in terms of grip strength, thermal pain threshold and mechanical pain threshold in three groups of mice before and after 10 weeks of intervention (D-I) Levels of serum IFN-γ, IL-10, RANKL, IL-17 A, TNF-α, and TGF-β after 10 weeks of behavioural monitoring. n = 12, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Total protein samples extracted from femoral tissue were incubated with primary and secondary antibodies for TNF-α (Abcam, USA, CAT: ab183218), RANKL (Proteintech, USA, CAT: ab239607), TGF-β1 (Abcam, USA, CAT: ab142139), and OPG(Abcam, USA, CAT: ab255723).

Techniques:

Journal: Scientific reports

Article Title: Black tea rich in polygonatum sibiricum polysaccharides inhibits osteoporosis in ovariectomized mice through immune regulation and gut microbiota suppression.

doi: 10.1038/s41598-025-08827-0

Figure Lengend Snippet: Fig. 2. (A-B) Changes in bone metabolism biomarkers, bone microstructure and related protein expression in three groups of mice after CBT intervention. (C-D) Representative images of ABH staining (40× scale bar: 1000 μm, 100× scale bar: 400 μm) (E) Analysis of the distal femoral epiphyseal region of mice using micro- computed tomography (CT). (F-G) Representative images of Immunohistochemistry of RANKL and OPG. (H-K) Comparison of BV/TV, Tb.Th, Tb.N, Tb.Sp. (L) Protein blot analysis of bone tissue targeting proteins associated with the RANKL/OPGpathway and inflammatory factors. (M-P) Expression of RANKL, TNF-α, OPG, and TGF-β1 proteins in bone tissues. n = 12, *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Total protein samples extracted from femoral tissue were incubated with primary and secondary antibodies for TNF-α (Abcam, USA, CAT: ab183218), RANKL (Proteintech, USA, CAT: ab239607), TGF-β1 (Abcam, USA, CAT: ab142139), and OPG(Abcam, USA, CAT: ab255723).

Techniques: Expressing, Staining, Micro-CT, Immunohistochemistry, Comparison

Journal: Scientific reports

Article Title: Black tea rich in polygonatum sibiricum polysaccharides inhibits osteoporosis in ovariectomized mice through immune regulation and gut microbiota suppression.

doi: 10.1038/s41598-025-08827-0

Figure Lengend Snippet: Fig. 6. (A-C) Spearman’s correlation analysis of gut microbiota at phylum, family and genus level with serum factors IFN-γ, IL-10, RANKL, IL-17 A, TNF-α and, TGF-β.(D) PICRUSt2 analysis identifies the top 10 KEGG pathways.

Article Snippet: Total protein samples extracted from femoral tissue were incubated with primary and secondary antibodies for TNF-α (Abcam, USA, CAT: ab183218), RANKL (Proteintech, USA, CAT: ab239607), TGF-β1 (Abcam, USA, CAT: ab142139), and OPG(Abcam, USA, CAT: ab255723).

Techniques:

Journal: Heliyon

Article Title: Coordination of early cellular reactions during activation of bone resorption in the rat mandible periosteum: An immunohistochemical study

doi: 10.1016/j.heliyon.2017.e00430

Figure Lengend Snippet: Osteogenic compartment. a and b : Immunostaining for RANKL. At baseline (Bs) a few immunopositive cells are scattered within the compartment. At 18 h ( b ), RANKL+ cells were more numerous. The staining was stronger than at baseline. Bar = 50 μm. c Changes with time in RANKL+ cells. d Variations in osteogenic cells and osteocytes expressing semaphorin3a. e and f Immunostaining for semaphorin 3a. At baseline most osteogenic cells were immunopositive as were the osteocytes (arrows) subjacent to the bone surface. At 24 h, only a few cells expressed the marker while the osteocytes no longer expressed it. Bar = 50 μm. * p ≤ 0.05; ** p<0.01; *** p<0.005 versus baseline. ° p<0.02; °° p<0.003 versus 18 h. # p<0.05 versus 24 h.

Article Snippet: Mouse monoclonal antibodies against HIS48 (ref 554905, BD Biosciences Pharmingen, San Diego, CA, USA: 1:100), ED1 (murine equivalent of CD68, MAB1435, Chemicon, Temecula, CA, USA; 1:100), CD54 (ICAM-1, ref 554967, BD Biosciences Pharmingen, 1:50) and rabbit polyclonal antibodies against CD106 (VCAM-1, SC-8304, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:50), RANKL (ref 3963, ProSci, Poway, CA, USA; 1/160), semaphorin 3a (ab23393, Abcam, Cambridge, UK; 1:50), laminin (L9393, Sigma; 1:25) were used.

Techniques: Immunostaining, Staining, Expressing, Marker

Journal: The British journal of nutrition

Article Title: Long-term resveratrol treatment prevents ovariectomy-induced osteopenia in rats without hyperplastic effects on the uterus.

doi: 10.1017/S0007114513003115

Figure Lengend Snippet: Fig. 5. Effect of resveratrol (Res) on the expressions of osteoprotegerin (OPG) and receptor activator of nuclear factor kB ligand (RANKL). After treatment, femur samples were collected and the expressions of OPG and RANKL were analysed by immunohistochemical methods. ((a)–(f)) Representative photomicrographs of femur sections showing OPG expression in the different groups : (a) the ovariectomy (OVX) group; (b–d) the Res 20, 40 and 80 mg/kg groups; (e) the oestradiol replacement therapy (ERT) group; (f) the Sham group. ((g)–(l)) Representative photomicrographs of femur sections showing RANKL expression: (g) the OVX group; ((h)–(j)) the Res 20, 40 and 80 mg/kg groups; (k) the ERT group; (l) the Sham group. Images were acquired at 200£ magnification. Arrows indicate the OPG- or RANKL-positive cells. (A colour version of this figure can be found online at http://journals.cambridge.org/bjn).

Article Snippet: H. Zhao et al.838 B ri ti sh Jo u rn al o f N u tr it io n niversity Press of rat IL-6, TNF-a, OPG and RANKL primary antibodies (Boster Bio-Engineering Company, Limited).

Techniques: Immunohistochemical staining, Expressing

Journal: Scientific Reports

Article Title: Accumulation of synovial fluid CD19 + CD24 hi CD27 + B cells was associated with bone destruction in rheumatoid arthritis

doi: 10.1038/s41598-020-71362-7

Figure Lengend Snippet: CD19 + CD24 hi CD27 + B cells in RA patients promoted osteoclast differentiation. Sorted RA SF CD19 + CD24 hi CD27 + B cells were cultured alone ( a ). Freshly isolated HC PBMCs (5 × 10 5 per well in 1 ml on a 12-well plate) were cultured in complete αMEM to allow adhering for 2 h in the incubator. Then non-adherent cells were removed to get the adherent osteoclast precursors (> 90% CD14 + ). After that, the adherent cells were cultured alone ( b ) or co-cultured with 1 × 10 5 OA SF CD19 + CD24 hi CD27 + B cells ( c ) or with RA PB 1 × 10 5 CD19 + CD24 hi CD27 + B cells ( d ) or with RA SF 1 × 10 5 CD19 + CD24 hi CD27 + B cells ( e ) in the presence of 30 ng rhM-CSF and 25 ng rhRANKL for 21 days or the addition of 100 ng anti RANKL antibodies for RANKL neutralization experiments ( f ) and stained for tartrate-resistant acid phosphatase (TRAP). TRAP positive multinucleate cells (more than three nuclei) were counted. Original magnification × 200. ( g ) Analysis of the number of TRAP+ multinucleate cells. All data were representative of three independent experiments. Statistical analyses between groups were performed using ANOVA. ** p < 0.01; NS, not significant.

Article Snippet: For RANKL neutralization experiments, 100 ng neutralizing antibody against human RANKL (Sino Biologicals) was added during the aforementioned co-culture system.

Techniques: Cell Culture, Isolation, Neutralization, Staining

Journal: Molecular Medicine Reports

Article Title: Effectiveness of combined salmon calcitonin and aspirin therapy for osteoporosis in ovariectomized rats

doi: 10.3892/mmr.2015.3637

Figure Lengend Snippet: Combined treatment (sCT+ASA) enhances the OPG/RANKL expression ratio in OVX rats. Effects of ASA, sCT and combined treatment (sCT+ASA) on mRNA levels in femur bone marrow cells from ovariectomized (OVX) rats. Gene expression was analyzed by reverse transcription-quantitative polymerase chain reaction. Values are expressed as the mean ± standard deviation. P-values were calculated by analysis of variance with Student-Newman-Keuls analysis. (A) OPG; (B) RANKL; (C) mRNA expression ratio of OPG/RANKL. ASA, aspirin; sCT, salmon calcitonin; OVX, ovariectomized; OPG, osteoprotegerin; RANKL, receptor activator of nuclear factor κB ligand; mRNA, messenger RNA.

Article Snippet: The antibodies used were as follows: OPG primary antibody (1:80; BA1475; Wuhan Boster Biological Technology Co. Ltd., Wuhan, China; rabbit anti-rat OPG), RANKL primary antibody (1:80; BA1323; Wuhan Boster Biological Technology Co. Ltd.; rabbit anti-rat OPG ligand) and rabbit anti-rat biotinylated secondary antibody (SP-0023; Beijing Biosynthesis Biotechnology Co., Ltd., Beijing, China).

Techniques: Expressing, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Effectiveness of combined salmon calcitonin and aspirin therapy for osteoporosis in ovariectomized rats

doi: 10.3892/mmr.2015.3637

Figure Lengend Snippet: Combined treatment significantly enhances the OPG/RANKL ratio in the proximal tibia of OVX rats. Effects of ASA, sCT and combined treatment (sCT+ASA) on protein expression levels in the proximal tibia of OVX rats. Specimens were observed at magnification, x250. Antigens were detected using an avidin-biotin-peroxidase system and visualized with 3,3′-diaminobenzidine. Protein levels were measured by densitometry. Values are expressed as the mean ± standard deviation. P-values were calculated by analysis of variance, with Student-Newman-Keuls analysis). (A) OPG; (B) RANKL; (C) OPG/RANKL protein ratio. Positive staining for OPG and RANKL was mainly localized in the chondrocytes of the epiphyseal growth plate and trabeculae of bone. ASA, aspirin; sCT, salmon calcitonin; OVX, ovariectomized; OPG, osteoprotegerin; RANKL, receptor activator of nuclear factor κB ligand.

Article Snippet: The antibodies used were as follows: OPG primary antibody (1:80; BA1475; Wuhan Boster Biological Technology Co. Ltd., Wuhan, China; rabbit anti-rat OPG), RANKL primary antibody (1:80; BA1323; Wuhan Boster Biological Technology Co. Ltd.; rabbit anti-rat OPG ligand) and rabbit anti-rat biotinylated secondary antibody (SP-0023; Beijing Biosynthesis Biotechnology Co., Ltd., Beijing, China).

Techniques: Expressing, Avidin-Biotin Assay, Standard Deviation, Staining