Anti-TRAF6 monoclonal antibody, clone FQ602Z

Purpose: Titanium particle-induced osteolysis is one of the important causes of aseptic loosening of artificial joints. Previous studies have shown the potential of natural compounds in preventing Ti particle-induced bone resorption. In this study, we observed the effects of magnesium lithospermate B (MLB) on titanium particle-induced osteoclast activity in vitro.Materials and Methods: RAW264.7 cells were treated with titanium particles (0.1 mg/mL) in the presence or absence of MLB (200nmol/L). We evaluated the osteoclast formation, bone pits formation and tartrate-resistant acid phosphatase 5b (Tracp5b) levels. Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to evaluate osteoclast differentiation-related genes (TRAF6, NFATc1, and c-fos) and protein expression.Results: The number of osteoclasts, pit formation and Tracp5b levels were all the group treated with titanium particles compared to the control group (all p <0.05). Titanium particles also promoted the expression of the TRAF6, NFATc1 and c-fos genes and protein expression. MLB significantly abolished the titanium particle-enhanced osteoclast and pits formation, and Traf6, NFATc1, and c-fos expression.Conclusions: Our data demonstrated that MLB can suppress titanium-induced osteoclast activity via inhibiting c-fos and NFATc1 expression.

Viral myocarditis (VMC) commonly triggers heart failure, for which no specific treatments are available. This study aims to explore the specific role of long non-coding RNA (lncRNA) maternally expressed 3 (MEG3) in VMC. A VMC mouse model was induced byCoxsackievirus B3(CVB3). Then, MEG3 and TNF receptor-associated factor 6 (TRAF6) were silenced and microRNA-223 (miR-223) was over-expressed in the VMC mice, followed by determination of ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS). Dual-luciferase reporter assay was introduced to test the interaction among MEG3, TRAF6 and miR-223. Macrophages were isolated from cardiac tissues and bone marrow, and polarization of M1 or M2 macrophages was induced. Then, the expressions of components of NLRP3 inflammatory body (NLRP3, ASC, Caspase-1), M1 markers (CD86, iNOS and TNF-alpha) and M2 markers (CD206, Arginase-1 and Fizz-1) were measured following MEG3 silencing. In the VMC mouse model, MEG3 and TRAF6 levels were obviously increased, while miR-223 expression was significantly reduced. Down-regulation of MEG3 resulted in the inhibition of TRAF6 by promoting miR-223. TRAF6 was negatively correlated with miR-223, but positively correlated with MEG3 expression. Down-regulations of MEG3 or TRAF6 or up-regulation of miR-223 was observed to increase mouse weight, survival rate, LVEF and LVFS, while inhibiting myocarditis and inflammationviathe NF-kappa B pathway inactivation in VMC mice. Down-regulation of MEG3 decreased M1 macrophage polarization and elevated M2 macrophage polarization by up-regulating miR-223. Collectively, down-regulation of MEG3 leads to the inhibition of inflammation and induces M2 macrophage polarizationviamiR-223/TRAF6/NF-kappa B axis, thus alleviating VMC.