The Effects Of Duantengyimu Decoction On The Expression Of Chemokine Receptor And Secretion Of Chemokine In Dendritic Cells Of Patients With Rheumatoid Arthritis

ObjectiveTo study the effect of Duantengyimu Decoction on the expression of chemokines receptors and secretion of chemokine in the peripheral blood dendritic cells from patients with rheumatoid arthritis,and to explore the mechanism of immunosuppression and treatment of rheumatoid arthritis about Duantengyimu Decoction on the migration of dendritic cells from the trend of dendritic cell.MethodThe mononuclear cells were isolated from the peripheral blood mononuclear cells of patients with rheumatoid arthritis by density gradient centrifugation.The monocytes were isolated by granulocyte macrophage colony stimulating factor and interleukin-4 for 7 days to differentiate.monocytes into Dendritic cells were treated with methotrexate(positive control group),or Duantengyimu Decoction lyophillization(test group),and then the levels of cytokines CXCL9 and CXCL10 in supernatant were measured by enzyme-linked immunosorbent assay and the expression levels of CCR5 and CCR7 on the surface of dendritic cells were detected by flow cytometry,and compared with the blank group.Result1.Under the common stimulation of GM-CSF and IL-4,peripheral blood mononuclear cells were successfully differentiated into DC,and typical dendritic morphology of DC was observed under light microscope.2.According to the results of CCK8 cytotoxicity and proliferation experiments,the optimal volume ratio of the dry powder of the Duantengyimu Decoction was 100mg/ml,it was obtained by comparing the A value of the different group and the normal group at the same time point.3.The results of ELISA showed that the levels of CXCL9 and CXCL10 of positive control group and test group in the supernatant were significantly decreased compared with the blank group(P<0.05).But there was no significant difference of CXCL9 and CXCL10 between the positive control group and test group(P>0.05).4.Flow cytometry showed that the expression of CCR5 and CCR7 on the DC surface after the Du Duantengyimu Decoction and methotrexate were significantly decreased,and the difference was statistically significant compared with blank group(P<0.05),the expression of CCR5 and CCR7 was slightly decreased(P>0.05),but the difference was not statistically significant compared with control group.Conclusion:The expression of CCR5 and CCR7 on the peripheral blood dendritic cells of patients with rheumatoid arthritis can reduce the secretion of chemokines CXCL9 and CXCL10,and thus inhibit the migration of dendritic cells to secondary lymphoid organs,Reduce its specific binding with T cells,which play a role in the treatment of rheumatoid arthritis.