|
Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease with a high incidence in Chinese people, which is characterized by synovitis. Synovitis and the serosa, heart, lung, skin, eyes, blood vessels and other diffuse inflammation of connective tissue are the main clinical manifestation, persistent,recurrent synovitis can lead to articular cartilage and bone destruction, joint dysfunction, severe limb deformities, even disability. Prevalence rate in China is about 0.4%. The cause of RA remains unclear,but genetic, environmental and immune factors may play a part role.It is the main principle of treatment that patients may obtain complete remission for a longtime.and the drugs are non-steroidal anti-inflammatory drugs, immunosuppressants, anti-rheumatic medicines, etc.But they could damage the function of liver and kidney, so we need to clear the cause of RA and find new therapeutic targets.As a new type of helper T cell subsets,it is the newly discovered CD4~+ T cell subsets,with an independent regulatory mechanisms of differentiation and development, can produce IL-17A, IL-17F and IL-22 and other cytokines . High levels of cytokine IL -17A is the markers of TH17 cell. Animal studies showed TH17 cells play an important role in autoimmune diseases and infectious diseases. At the same time there are some research indicates that a number of autoimmune diseases such as multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease and psoriasis have TH17 cells abnormality in quantity and function.This experiment studys of TH17 cells and its cytokine expression in the patient with rheumatoid arthritis and its clinical significance, may play a positive role in clarify the pathogenesis of RA and the development of new targeted therapies.Section I: The expression of TH17 cell related cytokines in RA patients’blood.Objective: Through detecting serum IL-17A protein, the expression levels of IL-17AmRNA in PBMC, this research described functional status of TH17 cell related cytokines in RA patients’blood.Methods: Activity group 32 patients and normal control group 30 people. Utilize Enzyme-linked immunosorbent assay and reverse transcription PCR method, detect IL-17A from protein and gene levels in RA patients blood respectively. Results: IL-17A expression in the serum of active RA patients was significantly higher than the normal control group(15.76±0.62pg/mlVS8.67±0.12pg/ml,P<0.05), IL-17AmRNA expression of PBMC in active RA patients was significantly higher than the normal control group(1.94±0.46VS0.84±0.16,P<0.05).IL-17A protein comtent was positively associated with Mrna expression level(r=0.7, P<0.05).Serum IL-17A level of RA patients and disease activity(DAS28)score were positively correlated(r=0.99, P<0.05).Conclusion: In active RA Patient, IL-17A levels were significantly elevated, and related to disease activity.Section II: the proportion of TH17 cells in RA patients’blood.Objective: IL-17 is a symbol cytokines of TH17 cell. By detecting the proportion of CD4 + IL-17 + T cell Reflect the quantity of Th17 cells in blood of RA patientsMethods: 32 RA patients were divided into activity group 32 patients and normal control group 30 patients. Ficoll density gradient centrifugation separate peripheral blood mononuclear cells (PBMC), a group without stimulation, the other group with phorbol ester and ionomycin stimulation for 4 hours, anti-CD4-PE and anti-IL-17-FITC labeled, Flow cytometry CD4~+IL-17+T cells ratio.Results: In RA patients’blood, CD4~+IL-17+T cells proportion was increased (2.50±0.54VS0.03±0.00,P<0.01), and there was no difference between stimulate and no stimulate group(3.23±0.05%VS2.50±0.54%, P﹥0.05). CD4~+IL-17+T cells of RA patients and disease activity(DAS28)score were positively correlated(r=0.96, P <0.05).Conclusion: the quantity of TH17 cells in RA patients Was increased and was related to disease activity.
|