| Objective: Rheumatoid arthritis (RA) was a kind of autoimmune disease characterized with arthritis and pannus. B lymphocyte differentiantion into plasma cell to produce rheumatoid factor and anti-cyclic cittrullinated peptides and other RA-related autoantibodies. These autoantibodies in the synovial membrane could rely on the role of complement to antigen-antibody complexes activate macrophages, resulting in the destruction of joints. It indicated that B lymphocytes played an important role in the pathogenesis of RA. T, B lymphocyte remained relatively balanced status through the regulatory mechanism, apoptosis. RA disease process can lead to abnormal apoptosis, while the Fas (CD95)/FasL system played an important role in regulation of apoptosis. The aim of this study was to investigate the expression of RA-specific antigen B lymphocytes CD20 and apoptotic factors CD95 in peripheral blood and the level of sFas/sFasL in serum and synovial fluid, to discussion B lymphocytes and apoptosis in the role of RA.Methods: 30 RA patients were chosen. The classification of all RA patients were consistent with American Rheumatism Institute and European congress prevention federation rheumatoid arthritis diagnostic criteria of 2009. All patients were previously untreated. Glucocorticoid (GC), disease modifying antirheumatic drugs (DMARDs) and biologic agent were never used for the RA patients. 20 healthy people as control were chosen. Serum was taken from all the patients and normal controls, synovial fluid was taken from 20 patients with RA. Clinic indexes of patients with rheumatoid arthritis were recorded such as age, disease duration, symptoms, the number of joint tenderness and swelling, DAS28, RF, ESR, CRP, CCP, IgA, IgM, IgG, C3, C4, etc. Avidin biotin peroxidase complex enzyme-linked immunosorbent assay (ABC-ELISA) was used to measure the levels of sFas/sFasL in serum and synovial fluid. Flow cytometric analysis (FCM) was employed for detecting apoptotic factors (CD95) of peripheral blood B lymphocytes cell (CD20). The correlation coefficients and significances were calculated between the percentage of CD20, CD95 and the laboratory parameters (RF, anti-CCP, ESR, CRP).All the data were analyzed by statistical software SPSS17.0 for windows. The mean number±standard deviation ( x±s) was used to express the measurement data. The test was adopted for comparison between groups. Chi-square test was used for the comparison of the enumeration data. Linear correlation was used to in analyse. P value<0.05 was considered significant.Results:1 The demographic details and traditional parameters of disease activity in RA: In the group of 30 patients with RA, the mean disease duration at presentation was (32.87±30.60) months, with a mean age of (48.73±14.86) yr, and 6 subjects were male, 24 subjects were female. In 20 normal controls, with a mean age of (45.00±11.08) yr, 5 subjects were male, 15 subjects were female. There were no differences between the patients and normal subjects in age, gender (P>0.05). In the group of 20 patients with RA, the mean ESR, CRP, RF and DAS28 were (67.03±29.88)mm/h, (32.06±22.68) mg/L, (413.58±463.72)IU/ml, (5.89±1.14).2 The sFas and sFasL levels in serum of RA patients: The sFas and sFasL levels in serum of RA patients were (3.78±1.02)μg/L and (0.77±0.41)μg/L which were significantly higher than normal group (2.68±0.81)μg/L and (0.36±0.21)μg/L. There were significant differences (P<0.01).3 The sFas and sFasL levels in synovial fluid of RA patients: The levels of sFas and sFasL in synovial fluid of RA patients, (5.62±1.4997)μg/L and (1.1935±0.4203)μg/L,were significantly higher than serum (P<0.01).4 The expression of peripheral blood CD20~+, CD95~+, and CD20~+/CD95~+ cells in RA: The expression of peripheral blood CD20~+, CD95~+, and CD20~+/CD95~+ cells in RA were (10.09±2.49)%, (41.35±7.63)%, (1.8±1.02)%. The expression of CD20+ cells in peripheral blood lymphocytes of RA group was significantly lower than normal controls (12.28±2.55)% (P<0.01). The expression of CD95+ cells in peripheral blood lymphocytes of RA group was significantly higher than normal controls (29.20±8.21)% (P<0.01). The expression of CD20+/CD95+ cells in peripheral blood lymphocytes of RA group was significantly lower than normal controls (2.43±1.11)% (P<0.01).5 The correlation between serum levels of sFas, sFasL and activity index of disease (DAS28, RF, ESR, CRP): According to correlation analysis, the levels of sFas in serum were positively correlated with ESR and CRP (r=0.76, P<0.01; r=0.84, P<0.01); The levels of sFasL in serum were positively correlated with ESR and CRP (r=0.81, P<0.01; r=0.54, P<0.01); sFas and sFasL were no found RF and DAS28 have obvious correlation (P>0.05).6 The positive rate of RA patients of cells surface expression of CD20+, CD95+and CD20+/CD95+ cells associated with the laboratory parameters of disease activity: According to correlation analysis, there was no significant correlation between the expression of CD20~+, CD95~+, CD20~+/CD95~+ cells activity index of disease (DAS28, ESR, CRP, RF) (P>0.05).Conclusions:1 The levels of sFas and sFasL in serum significantly increased in patients with RA, sFas and sFasL had correlation with laboratory parameters of disease activity (ESR, CRP), which could provide a disease activity index, monitoring of application of RA disease.2 There were higher levels of sFas and sFasL in synovial fluid than in serum, which suggested that sFas and sFasL might have relation to pathological changes of synovium of peripheral joint and articular cartilage destructions.3 The surface expression of CD20~+cells in peripheral blood lymphocytes of RA group was significantly lower than normal controls group, CD95~+cells in peripheral blood lymphocytes of RA group was significantly higher than normal group, CD20~+/CD95~+cells in peripheral blood lymphocytes of RA group was significantly lower than normal controls group, and it suggested that the abnormal changes of B lymphocyte and apoptotic factors of CD95~+ cells might had a relationship with the pathogenesis of RA.
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