Mechanism Of Qingluo Tongbi Granlue On Inhibiting Osteoclast Differentiation In Rheumatoid Arthritis

Objective:To discuss mechanism of Qingluo Tongbi Granlue on the basis of inhibiting osteoclast differentiation in rheumatoid arthritis through the methods of common cultivation of fibroblasts and mononuclear cells of arthritis rats to induce Osteoclast according to previous clinical and experimental work.Methods:Adjuvant arthritis rat model was made to extract and separate synovial fibroblasts and mononuclear cells, hanging small rooms and board with six orifices were used as training materials. These two types of cells were co-cultured (fibroblasts in the upper and six mononuclear cells in the lower) and made material exchange to simulate pathological process of rheumatoid arthritis and induce osteoclast, which was identified as tartrate resistant acid phosphatase (TRAP) staining positive multinucleated cells. Osteoclast successfully induced by this experimental has the function of bone resorption; Medicated serum with QLT was prepared in6groups, three holes for each group (repeat more than3times), which was added in co-culture system respectively at3-5d and18-21d, and the influence of TRAF6, ERK1/2, p38and JNK expression was observed through Western blot method48hours later. NFATcl expression was observed through immunofluorescence method; To further observe weather QLT plays regulating role on other pathway expression, MAPK pathway inhibitors was added on the basis of medicated serum with QLT to observe the differences of NFATcl expression among each group.Results:Medicated serum with QLT14.4,7.2,3.6g. k g-1could reduce osteoclast TRAF6, MAPK path and NFATcl expression in different degree, among which high and middle dose played obvious inhibitory action (P<0.01orP<0.05); The middle dose of QLT as the control group,added by MAPK pathway inhibitor, NFATcl expression had no significant difference compared with control group (P>0.05).Conclusion:Medicated serum with QLT could generally inhibit the expression of TRAF6, ERK1/2, p38, JNK signal pathways and NFATcl, and may regulate other signal paths which influence osteoclast differentiation and mature.