The Research Of Mechanism On Regulated Treg Cells Of Wen-yang-chu-shi Recipe On Collagen-in Duced Arthritis In DBA/1 Mice

Objective: To study the effects of Wen-Yang-Chu-Shi recipe(Juan-Bi granule)on the regulation mechanism of differentiation of Treg cells,analysis of “Juan-Bi granule in the treatment of Wang-Bi(cold dampness syndrome)” scientific connotation.Methods:(1)To construct the CIA model of bovine type II collagen induced by DBA/1 in mice with arthritis score,clinical score,average score of Juan-Bi granule alcohol extract group(given Juan-Bi granule alcohol extract 2.16mg/kg,equivalent to2.16g/kg crude drug),CIA model group(given the same volume of saline gavage),10 rats in each group;flow cytometry to detect the intracellular expression level in Treg cells;CIA mice of each group of paw joints and arthritis pathological damage were determined by H & E staining.(2)To construct cell model of i Treg cells induced differentiation in vitro.The level of Treg cytokines(TGF-??IL-10)was detected by ELISA,and the expression of intracellular CD4+Foxp3+ was detected by flow cytometry.We analyze the effect of Juan-Bi granule on the tropism of i Treg induced in vitro.(3)To construct Th17 cell model of Treg cells conversion in vitro,the expression of intracellular i Treg(CD4+ Foxp3)and Th17 cells(CD4+ IL-17+)was detected by flow cytometry.Analyze the effect of Juan-Bi granule on the plasticity of T cell differentiation.Result:(1)In Juan-Bi granule alcohol extract group,the arthritis symptoms in themice model group were significantly relieved(P<0.05),and Juan-Bi granule can reduce the local inflammatory cell infiltration in experimental mice,it can increase the proportion of Treg cells,also it can reduce clinical score.(2)In Juan-Bi granule alcohol extract group,the proportion of Treg cells increased,which significantly increased the ratio of Treg/Th17 cells(P<0.05).The alcohol extract of 50ug/ml Juan-Bi granule obviously promoted the level of TGF-? in the supernatant of i Treg in vitro,and the expression of specific cytokine and Specific transcription factor(Foxp3)of i Treg increased obviously,in which the difference between IL-10 and TGF-? was statistically significant(P<0.05).(3)Juan-Bi granule significantly inhibited the conversion of Treg cells to Th17 cells,and the difference was statistically significant(P<0.05),it indicated that Juan-Bi granule could block the transformation of Treg cells into pathogenic Th17 cells to a certain extent.Conclusion: Juan-Bi granule may prevent the differentiation of CD4+T cells plasticity highly pathogenic Th17 cells,it promotes the expression of key cytokines and transcription factors needed to the differentiation of Treg cells,and increases the proportion of Treg cells,also it can block transformation of differentiated Treg cells into hyperplastic cells Th17.These mechanisms maintain the dynamic balance of Treg/Th17,to achieve the purpose of treating Rheumatoid Arthritis.