The Research Of Orientvine Preparation Intervented The Incidence Of Rheumatoid Arthritis By Regulated T Follicular Helper Cells

Rheumatoid arthritis(RA)is an autoimmune disease characterized by the chronic synovitis in the joint,which involved in a variety of immune cells.The imbalance of immune cell subsets in RA patients and the imbalanced immune regulation of related cytokines are the hot spots in RA research field at present.The peripheral blood of most RA patients contain high levels of autoantibodies secreted by B cells,and the activation of B cells depends on the aid of T cells.T follicular helper(Tfh)cells plays an important auxiliary role in the activation and proliferation of B cells,which is a new T cell subset discovered after 2000.Tfh cells play an important bridge role in cellular immunity and humoral immunity,which is abnormality in the pathogenesis of RA.Therefore,the role of Tfh cells in the pathogenesis of RA is gradually taken seriously.The traditional Chinese medicine Orientvine is the dry rattan of the menispermaceae plant sinomenium acutum,which has been used for the treatment of inflammatory joint disease for thousands of years.There is no literature about intervention research of the Orientvine preparation in the rheumatoid arthritis Tfh cells.Introduction of Orientvine preparation into Tfh cells in the pathogenesis of RA will help to fill the blank of this filed.Objective:To observe the effect of Qingfengteng decoction(QFTD)combined with Methotrexate(MTX)on peripheral blood follicular helper Tfh cell,interleukin-21(IL-21),and disease activity in early rheumatoid arthritis patients;To observe the effect of Sinomenine on follicular helper T cell(Tfh)in collagen induced arthritis(CIA)mice,and to explore its possible mechanism for treating rheumatoid arthritis.Methods:Clinical trial:Totally 72 early RA patients were randomly assigned to the research group and control group,36 cases in each group.All patients took MTX 10mg once a week and celecoxib 0.2g bis in die,and the dosage of celecoxib was tapered according to the clinical symptoms.Patients in the research group additionally took QFTD.The treatment lasted for 12 weeks.Erythrocyte sedimentation rate(ESR)and Disease Activity Score in 28 joints(DAS28)were measured at the 0 week,4 weeks,8 weeks and 12 weeks.Tfh was detected by flow cytometry at 0 week and 12 weeks,IL-21 and Anti-cyclic citrullinated peptide antibody(CCP)were detected by enzyme linked immunosorbent assay(ELISA)at 0 week and 12 weeks.Experimental study:35 DBA/1 mice were randomly divided into model group(10 mice),sinomenine group(10 mice,74mg/kg·d)group,dexamethasone group(10 mice.0.88mg/kg-2d),and normal control group(5 mice).All the mice were administered intramuscular injected to the hindlimb.The injection lasted for 58 days.Scoring for CIA was performed after injection and arthritis index was calculated.All mice were killed after the last administration,and the joint pathology was observed.The peripheral blood Tfh cell ratio was determined by flow cytometry,and IL-21 was determined by ELISA.Spleen B-cell lymphoma-6(Bcl-6)mRNA expression were detected by Real-time Quantitative PCR Detecting System(qPCR),while spleen Bcl-6 protein were detected by western blot.Result:1.Compared with the same group before the treatment,ESR and DAS28 in the both groups decreased after the treatment(P<0.05,P<0.01).DAS28 of the research group was lower than that in the control group at 12 weeks(P<0.01).There was no significant difference about Anti-CCP before and after the treatment in both groups.2.Compared with the healthy population,the proportion of Tfh/CD4+T increased in both groups,as well as the level of IL-21(P<0.01).Compared with the same group before the treatment,the proportion of Tfh/CD4+T and the level of IL-21 decreased at 12 weeks,and which was lower in the research group than in the control group(P<0.01,P<0.05).3.Compared with the normal control group,the arthritis index obviously increased in the model group(P<0.01).Compared with the model group,the arthritis index obviously decreased in the sinomenine group and the dexamethasone group(P<0.01).4.Tfh cell ratio and IL-21 obviously increased in the model group than in the normal control group(P<0.01).Compared with the model group,the Tfh cell ratio and IL-21 obviously decreased in the sinomenine group(P<0.05,P<0.01),and the Tfh cell ratio decreased in the dexamethasone group(P<0.05).Compared with the dexamethasone group,the Tfh cell ratio and IL-21 decreased in the sinomenine group(P<0.05,P<0.01).5.Compared with the normal control group,the Bcl-6 mRNA expressions and the Bcl-6 protein in the model group were increased(P<0.01).Compared with the model group,the Bcl-6 mRNA expressions and the Bcl-6 protein in the sinomenine group were decreased(P<0.01).There were no significant difference about the Bcl-6 mRNA and the Bcl-6 protein between the model group and the dexamethasone group.Conclusion:QFTD combined with MTX could decrease the Tfh and IL-21 level in RA patients,and improve the clinical symptoms.Sinomenine could decrease the Bcl-6 mRNA expressions and Bcl-6 protein level,decrease the Tfh and IL-21 level in CIA mice,and improve the clinical symptoms.Orientvine preparation may regulate the onset of RA by intervent the Tfh cell.