Study On The Significance Of Peripheral Blood CD4~+CD25~+Foxp3~+ T Cells And Its Relationship With OPN In Patients With Rheumatoid Arthritis

Objective:By detecting the levels of Th17,Treg and OPN in peripheral blood of patients with rheumatoid arthritis(RA),to investigate(1)The number of Th17,CD4+CD25+Foxp3+T(Treg)cells and OPN content in peripheral blood of patients with RA;(2)The correlation between OPN content,Treg,Th17 cell count,the ratio of the two levels in peripheral blood of patients with RA and their association with the disease activity index;(3)The correlation between OPN content and Th17,Treg cell count and Th17/Treg,so as to find a possible pathogenesis related to the mechanism of RA,and new way for the treatment of RA.Methods:Cases that diagnosed with RA in first diagnosis were selected in Second Hospital of Shanxi Medical Collage in January 2016 and January 2017.There were 67 RA patients including 46 female cases,21 male cases,aging 29-84.And the average age was(57.5±12.0).Meanwhile,30 normal subjects as control group were selected from physical examination on Second Hospital of Shanxi Medical Collage.There were 21 female cases and 9 males,aging 33-79 years old.The average age was(55.3±16.8).Both groups were extracted 2 tubes of blood of Fasting vein in the next morning.One of the tube was used for detection of the number of Th17 and Treg by flow cytometry;the other was used for detection of OPN by enzyme-linked immunosorbent assay method(enzyme-linked immunosorbent assay ELISA).Meanwhile,The clinical data of RA group were collected,including gender,age,joint swelling,tenderness and so on.And Test parameters were anti-cyclic citrullinated peptide antibody(Anti-CCP),erythrocyte sedimentation rate(ESR),C reactive protein(CRP),rheumatoid factor(RF).At the same time,the DAS28 score of RA group was calculated.The changes of Th17,Treg and OPN between two groups were studied by correlation analysis.Results:1.Make Comparisons of the level of Th17,Treg and OPN in Peripheral blood between the RA and control groups.The numbers of Th17,Treg and levels of OPN in RA group were(7.64±6.44)/ul,(33.67±26.83)/ul,(14.84±8.64)ng/ml,while the control group were(6.64±4.26)/ul,(23.63±2.80)/ul,(10.59±3.40)ng/ml.Compared with the control group,Treg cells in group RA was significantly lower and OPN content increased significantly.the difference was statistically significant(P=0.001,P<0.001).Number of Th17 cells in RA tends to reduce(P= 0.192),and there was no statistical significance.The number of Th17/Treg of RA group is(0.32±0.34).Comparing with normal control group(0.28±0.18),the number of Th17/Treg trend to rise,there is no statistical significance(P=0.08).2.The number of Th17,Treg cell and correlation between Th17/Treg of 67 RA patients and clinical indexes:The number of Th17 was no significantly correlated with ESR,CRP,tenderness,swelling of the joints,DAS28 score,RF,Anti-CCP antibody(r=-0.162,P=0.191;r=-0.045,P=0.738;r=-0.235,P=0.056;r=-0.106,P=0.392;r=-0.153,P=0.289;r=0.875,P=0.574;r=0.170,P=0.224);The number of Treg was significantly negativel correlated with DAS28 score,tenderness joints,RF,(r=-0.317,P=0.009;r=-0.289,P=0.018;r=-0.411,P=0.003);The number of Treg was no significantly correlated The ESR,CRP,swelling of the joints,Anti-CCP antibody(r=-0.223,P=0.070;r=-0.124,P=0.352;r=-0.230,P=0.061;r=0.223,P=0.109).Th17/Treg was no significantly correlated with ESR,CRP,tenderness,swelling of the joints,DAS28 score,RF,Anti-CCP antibody(r=-0.046,P=0.710;r=-0.043,P=0.730;r=0.160,P=0.266;r=0.087,P=0.485;r=0.000,P=0.997;r=0.026,P=0.848;r=-0.016,P=0.896).3.Correlation between OPN levels of 67 RA patients and clinical indexes:OPN of RA patients was significantly positive correlated with ESR,DAS28 score,RF(r=0.284,P=0.02;r=0.290,P=0.039;r=0.376,P=0.007).There was no significant correlation between OPN and the CRP,tenderness joint,swollen joint(r=0.087,P=0.517;r=-0.105,P=0.399;r=-0.116,P=0.349).4.The level of OPN,the number of Th17,Treg in RA group and correlation between Th17,Treg: There is no significant correlation between OPN and Th17(r=0.006,P=0.963);Treg and OPN have a negativel correlation,and the difference was statistically significant(r=-0.459,P=0.000).There is no significant correlation between OPN and Th17/Treg ratio(r=0.202,P=0.102).Conclusion:1.The number of Treg cells in RA group was significantly lower than that in healthy control group,and was negatively correlated with disease activity;The number of Th17 cells did not change significantly,and there is no correlation with disease activity.As a kind of autoimmune disease,RA may be associated with the decrease of Treg cells,which can be used as a predictive index for evaluating RA disease activity.2.The levels of OPN in peripheral blood plasma in RA were significantly higher than healthy group,OPN was positively correlated with the laboratory parameters of disease activity.3.There was a negative correlation between Treg and OPN cells in RA group,which suggested that the reduction of Treg cell levels may be associated with elevated OPN concentration.