| Rheumatoid arthritis(RA)is an autoimmune disease characterized by changes in chronic arthritis,erosion of bone and cartilage,and hyperplasia of synovial tissue,and its pathogenesis has not been fully clarified.Moreover,the disease is easy to recur,the disease course is relatively long,the disability and teratogenic rate is relatively high,which causes a large economic and psychological burden on patients and families.Regulatory T cells(Tregs)are a subset of CD4+T cells that have significant inhibitory effects on immune effects.They can impede the immune response and immune response produced by other cells,maintain internal immune homeostasis,and prevent autoimmune diseases.And other aspects have a crucial role.In recent years,it has gradually become a research focus in the field of immunity.Studies have shown that there may be abnormalities in the number or function of Treg cells in RA patients,but the specific abnormal conditions and pathogenic mechanisms are not clear.ObjectiveTo evaluate the function of regulatory T(Treg)cells in peripheral blood from patients with rheumatoid arthritis,and to explore the possible role of the STAT3 signaling pathway in Treg cell dysfunction.MethodsTotally,60 patients with rheumatoid arthritis,were enrolled into this study.sixty healthy blood donors served as the control group.Venous blood samples were collected from these subjects followed by isolation of Treg cells and responder T(Tresp)cells.Flowcytometry was performed to determine the proportion of Treg cells in peripheral blood as well as that of cells secreting phosphorylated-STAT3(p-STAT3),interferonγ(IFN-γ),tumor necrosis factor α(TNF-α)and interleukin 17A(IL-17A)in Treg cells,and quantitative real-time PCR(qRT-PCR)to measure the expression levels of IFN-y,TNF-α and IL-17mRNAs in Treg cells.Some Treg cells and Tresp cells were cultured in vitro alone or in combination,and flow cytometry was conducted to estimate cellular proliferative activity on day 7 after stimulation with IL-2.Some patient-derived Treg cells were classified into several groups to be cultured alone or in combination with Tresp cells with or without the presence of the STAT3 pathway inhibitor,Stattic V(10 or 50 μg/L),for 7 days.Subsequently,flow cytometry was performed to evaluate the proliferative activity of Tresp cells,and qRT-PCR to measure the expression levels of IFN-γ,TNF-α and IL-17A mRNAs in Treg cells.Results(1)The number of Treg cells in the peripheral blood of the RA patient group(6.51%±0.24%)was significantly different from that in the normal control group(2.23%±0.18%)(t=4.22,P<0.05).(2)The proliferation activity of Treg cells in peripheral blood and the inhibitory function of Tresp cells in patients with RA were significantly reduced(t was 4.23 and 4.35,both P<0.05),and the expression level of phosphorylated STAT3 was significantly increased(t=3.84,P<0.05),the levels of secreted pro-inflammatory factors IFN-γ,TNF-α,and IL-17A increased significantly(t were 3.61,2.51,and 2.69,all P<0.05).(3)After the effect of 50 μg/L StatticV,the Treg inhibition rate of Treg cells in RA patients was(61.24%± 4.62%),and the untreated group was(28.15%±10.37%).The difference between the two groups was statistically significant(P<0.05),the expression levels of pro-inflammatory factors IFN-γ,TNF-α,IL-17AmRNA(2-ΔΔCt)were(1.65 ± 0.88,0.85 ± 0.71,0.57 ± 0.14),which were significantly lower than those in the untreated group Were(23.32 ± 6.71,4.85 ± 1.53,3.10 ± 0.62),and the differences were statistically significant(all P<0.05).ConclusionsThe number and function of Treg cells in RA patients are abnormal,and the inhibitory function of Tresp cells is reduced,leading to disturbances in the secretion of related cytokines.The mechanism of immune disorders is closely related to the abnormal activation of STAT3 signal transduction pathways,which inhibits the abnormality of STAT3 signal transduction pathways.Activation plays an important role in improving and restoring Treg cell function. |
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