Study On The Effect Of Qingluo Tongbi Granule On Rheumatoid Arthritis Related Differentiation Factor Of Osteoclast

Objective:On the basis of effective treatment RA,the target is to further explore the effect of Qingluo Tongbi Granule on rheumatoid arthritis related differentiation factor of osteoclast from the molecular level,for further reveal Qingluo Tongbi Granule on rheumatoid arthritis signal path mechanism of osteoclast lay the foundation.Method:This experiment used peripheral blood mononuclear cells in co-culture method, set up a synovial mechanocyte-mononuclear cell training together model. When arthritis rats synovial fibroblasts grow to3-5generation,digest them from the cell bottle,with15%fetal calf serum MEM adjust the concentration to0.5-1.0*106/ml cell suspension liquid set aside,the transwell contained mononuclear cells suspension liquid six orifice cell plate at the bottom of the plate,and then put mechanocyte cell suspension into the transwell. Buy5percent,carbon dioxide incubator37centigrade training,with M-CSF as inducers and induce the formation with typical osteoclast characteristics of osteoclast cell,and identify with TRAP dyeing.In the microscopic:cells form clear,an irregular form,more nuclear (about4-10),sappanwood staining red or palm red part of the TRAP for enzyme activity in parts, which identified as osteoclast meet the appraisal standards. The experiments prove the establishment of the system can be trained induction of success with bone absorption function of the osteoclast. Trained to the third,the eleventh the twenty first day,give the medicine to six groups,each group set three parallel comparison,training after48hours then collect the cell clear liquid,used for RANKL, IL-34and OPN detection.Put the cell clear liquid into ninty-six orifice cell plate,detect the content about RANKL,IL-34and OPN,each group make four parallel comparison,methods follow the each kit instruction book. Determine the effect on the containing medicine serum to RA osteoclast differentiation factor RANKL and broken bone cells related differentiation factor IL-34and OPN.Results:Regular doses of QLT medicated serum can restrain the content of osteoclast cell serum RANKL, IL-34and OPN,can restrain osteoclast proliferation,and make osteoclast differentiation,abate proliferation activity,it shows that QLT osteoclast growth to a direct inhibition.Conclusion:Regular doses of QLT by suppressing the RANKL upstream of inflammatory cells secrete factors (IL-34,OPN etc), thus mediated the OC RANKL differentiation of the key factors in excessive expression, reduced the RA articular cartilage and subchondral bone destruction, and slow down the process of bone destruction.