TAZ Activates Th17 Cells In Ankylosing Spondylitis And Intervention Mechanism Of Triptolide

Background:Ankylosing spondylitis(AS)is a common inflammatory spondyloarthropathy in clinic.It is characterized by chronic inflammation of sacroiliac joint and spinal attachment point.It can also involve peripheral joints and is often accompanied by various degrees of systemic multisystem injury.This disease often involves young men who are in the best state of brain and physical labor.In the late stage,ankylosis deformity occurs,which seriously reduces the quality of life of patients.Inflammation is the starting link of pathological changes in AS.Once inflammation occurs,it will cause pathological new bone formation in AS patients,make the pathological changes into an irreversible period,and eventually lead to disability.Therefore,the control of inflammation is the focus of AS treatment.T helper cell 17(Th17)is derived from pluripotent CD4+T cells and can play an inflammatory role in secreting a large amount of cytokine interleukin 17(IL-17).Th17/IL-17 inflammatory axis plays an important role in the development of AS inflammation.Inhibiting the differentiation of inflammatory Th 17 has become a key therapeutic target for AS,TAZ(transcriptional co activator with PDZ binding motif)is a key transcriptional coactivator in Th17 cells.After antigen stimulation,initial CD4+T cells are activated,expanded and differentiated into Th1,Th2,Th17 and regulatory T cell(Treg)subsets.The differentiation process is regulated by different cytokines and characteristic transcription factors.Different from Th1,Th2 and Treg subgroups,RORγt is a specific transcription factor that controls the differentiation of Th17 cells.Transcription factors can not play a role alone in the process of gene expression,so they need to cooperate with transcriptional coactivators to form transcription complexes to regulate gene expression.Studies have shown that the transcription coactivator TAZ can interact with RORyt forms transcription complex and enhances RORyt transcriptional activity regulates the differentiation of inflammatory Th17.Therefore,to explore the expression of TAZ in patients with ankylosing spondylitis and its effect on Th17 differentiation in AS patients is of great significance for alleviating and controlling AS inflammation.The pathogenesis of AS is based on the deficiency of kidney and governor vessel,and marked by wind,cold,dampness and heat.Among them,damp heat obstruction is a common clinical syndrome type of AS,which should be treated with the method of promoting dampness and clearing heat.Tripterygium wilfordii is the root,leaf,flower and fruit of Tripterygium Wilfordii in Celastraceae.It is good at dispelling wind and removing dampness,clearing heat and activating blood circulation,dredging collaterals and relieving pain.Because of its strong anti-inflammatory and immunosuppressive effects,Tripterygium Wilfordii and its preparations have been widely used in the treatment of inflammatory diseases and autoimmune diseases.Network pharmacology studies show that the active ingredients of Tripterygium wilfordii may play a role in the treatment of as through Th17 differentiation pathway.Triptolide is the main active ingredient of Tripterygium Wilfordii.Previous studies of our group showed that triptolide can effectively interfere with Th17 differentiation and IL-17 secretion in peripheral blood of AS patients.Based on the above research,this study will continue to focus on Th17 differentiation,and explore the expression of TAZ in AS patients and the intervention effect of triptolide.Objective:To explore the expression of transcriptional co-activation factor TAZ in a single nuclear cell(peria blood cell,PBMC)in AS patients,and its effect on the regulation of Th17 cell differentiation,and the effect of refractory on the expression levels of RORyt and TAZ in active patients.In order to find a new therapeutic target for AS and provide a new direction for new drug research and development.Methods:A total of 50 AS patients were included in the study,including 25 AS active patients and 25 AS stable patients;there were 20 healthy controls.After taking the elbow nest vein blood from each group,individual nucleocells of serum and exosymetic blood were collected using Ficoll-hypaque density gradient centrifugal method(external blood mononuclear cell,PBMC);The protein expression levels of TAZ and RORγt in each group of PBMC were detected by the protein immunoprinting method(western blot,WB).At the same time,the correlation between TAZ expression level and RORγt expression level,serum IL-17 expression level and inflammatory index ESR and CRP were analyzed.Study Ⅱ included 25 cases of AS patients in the active period,and PBMC in the in vitro intervention period of Chinese medicine monogamline retocin,to observe the effects of triptolide on the expression of RORyt and TAZ.Using SPSS 25.0 statistical analysis.All tests are two-sided,with measurement data expressed in x±s,and the difference is considered statistically significant when the p-value is<0.05.Comparing the groups,both sets of data conform to the normal distribution and meet the variance of the two groups of independent samples t-test;For those who do not conform to normal distributions or variances,the non-parameter Kruskal-Wallis H test is used.The two sets of measurement data conform to the normal distribution,using Pearson correlation coefficient for correlation analysis,and not meeting the normal distribution,using Spearman for correlation analysis.Results:1.The serum IL-17 expression level of AS patients in the active period increased significantly(p<0.05)compared to the normal control group(p<0.01),and the serum IL-17 expression level in stable AS patients increased significantly compared to the normal group,but was not statistically significant(p>0.05).2.The expression of RORγt protein in AS patients during the active period was significantly increased during the stable period and in the healthy control group(p<0.01),and the expression of RORγt protein in stable period was significantly higher than in normal people(p<0.01).3.Compared with the healthy control group,TAZ protein expression was significantly increased in AS patients during the active period(p<0.01),and in the active period AS patients with TAZ protein expression was significantly increased(p<0.01)compared to the stabilization period.The expression of TAZ protein in patients with stable AS was significantly higher than that in normal subjects(p<0.01).4.After correlation analysis,it is found that the expression level of TAZ is significantly related to RORγt(r=0.730;p<0.01)and significantly related to ESR and CRP(TAZ-ESR:r=0.603;p=0.001<0.05;TAZ-CRP:r=0.416;p=0.039<0.05),significantly related to serum IL-17 levels(r=0.800;p<0.010).There was no significant correlation between TAZ expression level and the disease activity index BASDAI score(r=0.353;p=0.084>0.05).5.After in vitro intervention of Chinese medicine triptolide,the expression of RORγt protein in PBMC in AS patients during the active period was significantly reduced(p<0.01)and the expression of TAZ protein was significantly reduced compared to pre-intervention(p<0.01).Conclusions:1.The expression of TAZ in PBMC of AS patients was significantly increased,which may be related to RORyt forms a transcription complex,which positively regulates the differentiation of inflammatory Th 17 and participates in the pathogenesis of AS.2.After the vitro intervention of triptolide,the expression of RORyt and TAZ are significantly reduced in PBMC of AS patients,which may be an effective therapeutic drug for intervention of Th17 differentiation in AS patients,thereby alleviating inflammation.