Immunomodulatory effects of berberine on Staphylococcus aureus-induced septic arthritis through down-regulation of Th17 and Treg signaling pathways.

OBJECTIVES: The preseaant study was aimed to investigate the immunomodulatory effects of berberine on Staphylococcus aureus-induced septic arthritis through the downstream signaling mechanism of Th17 and Treg, in the control and prevention of disease progression of Staphylococcus aureus induced septic arthritis of blood, spleen and synovial joints. METHODS: The study was conducted in mice induced with septic arthritis by S. aureus for 15 days. The infected mice were treated with berberine (50 or 100 or 200 mg/Kg) to evaluate the effects on the isolated cells of Th17 and Treg from synovial joints, blood and spleen against the septic arthritic induced mice followed by JNK, RANKL and NF-κB expressions in the lysates of Th17 and Tregs isolated cells. The evaluation of serum IL-21 and TGF-ß levels was also conducted after 15 days post-infection in Th17 and Treg population. RESULTS: Our findings showed that berberine exerted excellent inhibitory effects on the S. aureus (AS-789) strain for inducing sepsis-induced arthritis. The results from the S. aureus testing revealed that at concentrations below 640 µg/mL, the strain was more resistant to berberine, as it had an increased rate of growth. The assessment of S. aureus induced septic arthritis (joint swelling and arthritis index) substantial reduction in the joint swelling and arthritis index (p<0.01) in the berberine-treated groups. The percentage of Th17 cells with CD4 and RORγt; Treg cells with CD4, CD25 and FOXp3 in the synovial joints, blood and spleen was substantially declined in the drug-treated groups (p<0.01) as compared to the S. aureus infected mice. The TGF-ß and IL-21 serum levels determinations in S. aureus induced septic arthritis revealed a substantial decrease in serum TGF-ß levels (p<0.01) in drug-treated groups compared to the infected animals. The post hoc test revealed a substantial decrease in JNK, NF-κB and RANKL expressions in the lysates of Th17 and Treg isolated cells in the drug-treated animals (p<0.01) when compared to the S. aureus-infected cluster. CONCLUSION: Our findings demonstrated that a possible strategy for combating disease severity with berberine treatment in Staphylococcus aureus induced septic arthritis in mice, which targets the Th17 and Treg cells have driven the NF-κB/JNK-RANKL axis.

Osteopontin-induced lncRNA HOTAIR expression is involved in osteoarthritis by regulating cell proliferation.

BACKGROUND: Osteopontin plays critical roles in osteoarthritis (OA) by regulating the functions of osteoclasts. It is known that osteopontin can induce the expression of lncRNA HOX transcript antisense RNA (HOTAIR), indicating the involvement of HOTAIR in OA. This study was carried out to investigate the role of HOTAIR in OA. METHODS: Synovial fluid was extracted from both OA patients (n = 58) and healthy controls (n = 58). Expression of osteopontin and HOTAIR in synovial fluid was determined by RT-qPCR. Osteopontin was used to treat chondrocytes at dosages of 0, 1, 5 and 10 µg/ml, followed by measurement of HOTAIR expression by RT-qPCR. The role of osteopontin and HOTAIR overexpression, as well as HOTAIR knockdown in regulating the proliferation of chondrocytes was analyzed by cck-8 assay. RESULTS: HOTAIR was upregulated in OA. A positive correlation between HOTAIR and osteopontin was observed. In the primary chondrocytes, osteopontin treatment increased HOTAIR expression, while HOTAIR overexpression and knockdown failed to significantly affect osteopontin expression. In addition, osteopontin and HOTAIR overexpression increased chondrocyte proliferation, while HOTAIRE knockdown decreased chondrocyte proliferation. In addition, HOTAIR knockdown reduced the effects of osteopontin treatment on cell proliferation. CONCLUSIONS: Osteopontin-induced HOTAIR expression is involved in osteoarthritis by regulating cell proliferation.

SNHG16/miR-485-5p/BMP7 axis modulates osteogenic differentiation of human bone marrow-derived mesenchymal stem cells.

BACKGROUND: Osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) is crucial for bone formation and its dysfunction is reported to be linked to osteoporosis (OP). The present study aimed to probe the function of the long non-coding RNA small nucleolar RNA host gene 16 (SNHG16) with respect to modulating the osteogenic differentiation of hBMSCs. METHODS: SNHG16 expression in hBMSCs obtained from OP patients was measured by a quantitative real-time polymerase chain reaction (qRT-PCR). Gain-of-function and loss-of-function models of SNHG16 were established with hBMSCs. The expression of OP-related genes (ALP, OCN and OPN) in hBMSCs was determined by qRT-PCR and western blotting. StarBase, TargetScan7.2, miRDB and PicTar databases were used to predict the binding sites between SNHG16 and miR-485-5p, miR-485-5p and 3'-UTR of bone morphogenetic protein 7 (BMP7), respectively. A dual-luciferase reporter assay was used to determine the regulatory relationships between SNHG16 and miR-485-5p, miR-485-5p and 3'-UTR of BMP7, respectively. RESULTS: SNHG16 was remarkably down-regulated in hBMSCs obtained from patients with OP. Overexpression of SNHG16 promoted the osteogenic differentiation of hBMSCs, whereas knockdown of SNHG16 suppressed it. Mechanistically, miR-485-5p is a target of SNHG16, and miR-485-5p can reverse the function of SNHG16. BMP7 is also identified as a target of miR-485-5p and can be indirectly modulated by SNHG16 in hBMSCs. CONCLUSIONS: SNHG16 promotes the osteogenic differentiation of hBMSCs via regulating the miR-485-5p/BMP7 axis and comprises a prospective therapy target for OP.

Disease perception and awareness investigation of secondary osteoporosis in patients with chronic obstructive pulmonary in Urumqi / 国际中医中药杂志

ObjectiveTo evaluate perception and awareness of disease knowledge in patients with chronic obstructive pulmonary disease (COPD) in rural area of Urumqi city.MethodsA survey was made to collect 121 patients with chronic obstructive pulmonary disease in out-patient and hospitalization of Traditional Chinese Medicine Hospital of Xinjiang Medical University in 2011.Questionnaire data were coded and entered into Microsoft EpiDate by two persons independently,with SPSS 17.0 version statistical package was used for statistical analysis.ResultsAmong 121 sampling subjects,104 participants completed the questionnaire qualifiedly.Among these patients,Male occupied 65.4％ (68/104),female occupied 34.6％ (36/104).Patients who know that post-menopausal women were easier to suffer osteoporosis totaled 42 cases (40.38％),patients who know that bedridden persons were easier to suffer osteoporosis totaled 42 cases (40.38％),patients who know that female was more likely to suffer than male totaled 65 cases (62.5％),who think the disease is related to the gender totaled 76 cases (73.08％),who think it is unlikely to develop osteoporosis after fracture healing totaled 13 cases (12.5％).Patients who do not think hat low back pain is the common manifestation of osteoporosis totaled 23 cases (22.12％),Patients who do not think hump and fracture are the common osteoporosis performance totaled 14 cases (13.46％) and 4 cases (3.85％) ; patients who know that osteoporosis is often accompanied by bone spurs totaled 33 cases (31.3％),patients who know that bone mineral density is a reasonable detection of osteoporosis totaled 62 cases (59.61％).Patients who know about alendronate phosphate (such as Fosamax,etc.) totaled 12 cases (11.76％),patients who know that commonly used TCM medicines for treating osteoporosis are Eucommia and Epimedium totaled 5 cases (4.90％).ConclusionCOPD is prone to cause osteoporosis,but patients are lacking the awareness of osteoporosis.

Construction and pathological features of a rabbit osteoarthritis model in early, middle, and later phases / 中国组织工程研究

BACKGROUND: Application of traditional osteoarthritis animal models was limited by long duplicated time, poor stability, different successful rate, and rough analysis of osteoarthritis.OBJECTIVE: To observe clinical and pathological features of osteoarthritis by modified Hulth modeling way and to determine the stages of ostecarthritis.METHODS: A 2-cm medial longitudinal incision was resected to expose knee joint. Anterior and posterior cruciate ligament and medial collateral ligament were cut off, and medial meniscus was fully cut to reserve articular cartilage., The injured limb was not fixed postoperatively. Animals were allowed to move freely. At one week after surgery, 800 000 U penicillin was used to avoid from infection, 30 mind, twice per day, for 12 successive weeks. The normal group was treated without any treatments. Pathological features were observed using Mankin scores under electron and optic microscope at 4, 6, 8, 10, and 12 weeks postoperatively.RESULTS AND CONCLUSION: Changes of osteoarthritis were observed in the model group at 4 and 6 weeks after operation,showing synovial hyperemia and hyperplasia, increased synovial fluid effusion, cartilage surface roughness, matrix stained tinge,and Mankin score of 3.5-3.8. Intermediate stage changes of osteoarthdUs were found in the model group at 8 weeks after operation, showing synovial hyperplasia, less synovial fluid, flssuration reaching cartilage surface, cartilage cells with tangled and uneven staining matrix, Mankins score of 8-9. Advanced osteoarthritis changes were observed in the model group at 12 weeks after operation, showing severe nodular synovial hyperplasia, less and turbid synovial fluid, osteophyte formation of serious exposure of subchondral bone, cartilage calls reducing the majority of loss of matrix staining, and Mankin score of 12-14. Electron microscopy indicated a coincidence with the histological observation of cell mutation. The rabbit model by Hulth suggested that early change of osteoarthritis occurred at 6 weeks after operation, intermediate stage at 8 weeks, and advanced stage at 12 weeks.This model could be more comprehensive response to osteoarthdtis cartilage degeneration from early compensatory hypertrophy to decompensation after the cartilage cells and matrix reduction, cartilage softened to endarterectomy missing characterized the late changes in the entire process.