| ObjectiveTo investigate the differentiation of T helper cells and its related cytokines’ impact on rheumatoid arthritis progress.study the correlation analysis between dendritic cell subsets/SIRT1mRNA and Th cells differentiation in peripheral blood in patients with RA, which may clue a new target therapy, which illuminates deeply the pathogenesis of rheumatoid arthritis.MethodsA total of94RA patients and22normal controls (NCs) were enrolled. All the RA patients into this study were divided into different disease activity groups according to Disease Activity Score. The frequency of lymphocyte subsets, dendritic cells (mDC, pDC), Th1/Th2cells and Th17/Treg cells in peripheral blood was evaluated by fow cytometry (FACS). Real-time fluorescent quantitative PCR (RQ-PCR) was used to measure the expression of RORγt gene, FoxP3mRNA and SIRT1mRNA in PBMC. The concentrations of IFN-γ, IL-4, IL-17, TGF-β1in serum were measured by enzyme-linked immunosorbent assay (ELISA). The correlation analysis was studied between cytokines and DAS28activity index or laboratory inflammation indexs (ESR, hsCRP, RF, anti-CCP).Results1. There was no signifieant difference of CD3+T cells between RA patients and NCs. A significant increase in the frequency of CD3+CD4+T cells and CD4+/CD8+ratio（p<0.05）, while a significant decrease in the frequency of CD3+CD8+T cells in peripheral blood of RA patients compared with normal controls（p<0.05）. Followed with the elevation of DAS28activity index, the frequency of CD3+CD4+T cells and CD4+/CD8+ratios had a little increase, but it was no signifieant difference（p>0.05）.2. Compared with NCs, the frequency of Th1cells and Th17cells were significantly increased in RA patients（p<0.05）. Th1cells had a sharp elevation in high active RA patients（p<0.001）. As the the elevation of DAS28activity index, the frequency of Th1cells had a little increase among different active RA groups, but there was no obvious difference（p>0.05）. The percentage of Th2cells had no significant difference between RA patients and NCs（p>0.05）. The frequency of Treg cells were significantly decreased（p<0.001）, Th1/Th2ratios showed the trend of increase in RA patients, but no obvious change was observed among different active patients（p>0.05）. Th17/Treg ratios were significantly increased in high active patients（p<0.001）, while no significant difference among other groups（p>0.05）. Th17/Treg ratios were positively correlated with serum ESR, hsCRP and DAS28activity index（p<0.05）, but negatively correlated with Th1/Th2ratios（p<0.05）.3Compared with mild, middle active patients and NCs, the expression of RORγt mRNA levels in high active patients increased（p<0.05）, but the expression of FoxP3mRNA levels decreased（p<0.05）.4. Compared with NCs, the levels of IFN-γ and IL-17were increase in RA patient（sp<0.05）, serum IFN-γ levels had a sharp elevation in high active RA patients（p<0.001）. As the the elevation of DAS28activity index, serum IL-17levels had a significant increase. Serum IL-4levels were significantly decreased in middle active and high active patients（p<0.001）. Serum TGF-β levels were obviously decreased in mild active and middle active patients（p<0.01）, but increased in high active patients（p<0.05）. IFN-γ/IL-4ratios were significantly higher than NCs（p<0.01）, followed with the elevation of DAS28activity index. IFN-γ/IL-4ratios had obvious increase in RA patients. Serum IL-17and TGF-β levels were positively correlated with ESR, hsCRP and DAS28activity index（p<0.05）.5. The number of mDC in peripheral blood of high active RA patients was significantly decreased（p<0.01）, while no obvious difference was foud among other group（sp>0.05）. pDC number were clearly lower in RA patients than NC（sp<0.001）, which had the most decrease in high active patients. But mDC/pDC ratios were higher in RA patients than NCs（p<0.05）.6. The number of mDC in peripheral blood of RA patients was negatively correlated with Th17cells, Th17/Treg ratios, IFN-γ and IL-17levels（p<0.05）. The number of pDC was positively correlated with Treg cells（p<0.01）, but negatively correlated with Th17/Treg ratios, IFN-γ, IL-17and TGF-β levels（p<0.05）.There was a positive correlation between mDC/pDC ratios and Th17cells or Treg cell（sp<0.05）.7. Compared with mild, middle active patients and NCs, the expression of SIRT1mRNA levels in PBMC of high active patients decreased（p<0.05）, The SIRT1mRNA levels were negatively correlated with Th1cells, Th1/Th2ratios, IFN-γ and IL-17（p<0.05）, but positively correlated with Th2cells and RORγt, FoxP3mRNA levels（p<0.05）.Conclusion1. There would be Th1/Th2and Thl7/Treg imbalance in peripheral blood of RA patients, mainly Th17/Treg imbalance, which may be involed in pathogenesis of RA patients.2The number of DC subset is abnormal during the course of RA that probably result in Th17/Treg and cytokines environment imbalances.3. SIRT1may delay the pathogenesis of RA by affecting Th cells differentiation and secretion of cytokines. |
