| Objective Rheumatoid arthritis (RA) is a highly disabling disease, combination is still a major treatment method, and many common combination therapy at international at present. The effect different treatment therapy in RA patients has been clear, it is unknown that the relationship between clinical routine treatment effect and the expression of Thl7/Treg. Clinical routine treatment such as methotrexate(MTX), Sulfasalazin (SSZ),chloroquin (HCQ), corticosteroids and anti-TNF alpha, whether can they adjust Th17/Treg balance and which kind of scheme can better regulate Thl7/Treg balance,it has not been decided yet, and there are many differences between the animal experiment and clinical trials. This study is to understand the expression of IL-17in peripheral blood by clinical conventional treatment, to make the relationship definitely between the clinical routine treatment effect and the expression of Thl7/Treg, in order to formulate more effective treatments to provide the basis. Our study will develope RA animal model, in order to elucidate the mechanism of combination therapy. At home and abroad, there has not been seen the similar reports, for the treatment of RA providing new ideas and methods. By researching collagen induced arthritis (CIA), from the angle of inflammatory cytokines, T lymphocytes balance and apoptosis, to study the mechanism of clinical routine treatment.Methods To established CIA on male wistar rats successfully, then they were randomly divided into CIA model group, MTX (2.7mg/kg/w)+prednisone (5mg/kg/day)group, MTX (2.7mg/kg/w)+chloroquin(40mg/kg/day)+Sulfasalazin (160mg/kg/day) group, MTX(2.7mg/kg/w)+Etanercept (0.3mg/kg/day)group.3weeks after the first immunization, we began to medicine. Treatment effects were assessed by clinically, histologically and radiologically. Clinical evaluations of each paw welling were assessed by arthritis index(AI).21weeks after the first immunization, all of the rats were sacrificed, ankle joints were exposed,fixed, decalcified,wrapped and cut into slices. All slices were stained with hematoxylin and eosin to evaluate synovial inflammation and bone erosive changes, further:A. The serum IL-17level were test by ELISA; B. flow cytometry detection the number of Th17and Treg and their apoptosis.Results1. The rate of rat developed CIA model was90.3%.2. Arthritis index (AI):The treatment groups are more improved obviously than CIA group, the AI of MTX with prednisone group was most lowest in the three treatment groups (P<0.05), and it improve the swelling degree of ankle was most obvious (P<0.05). 3. HE ankle pathology:In CIA group, the synovial hyperplasia, arranging disorderly, a large number of inflammatory cells infiltration, cartilage and bone tissue serious erosion; In MTX with chloroquin and Sulfasalazin group synovial inflammation, cartilage and bone tissue all ease erosion; In MTX with prednisone group and MTX with Etanercept group synovial inflammation, cartilage and bone tissue erosion obviously improved.4. In MTX with prednisone group, the weights (P<0.05)was lighter than other group.5. In CIA model group, serum IL-17level (P<0.05)was higher than normal group, each treatment group all inhibited IL-17level.6. Th17and Treg cells subsets:(1)10weeks after immunization, Treg cells increased and apoptosis reduced in the treatment groups than the CIA model group,which showed statistically significant (P<0.05).(2)21weeks after immunization, Thl7cells in three treatment groups were reduced than CIA model group, which showed statistically significant (P<0.05). In MTX+chloroquin+Sulfasalazin group,Th17cells were less than MTX with prednisone group, which showed statistically significant (P<0.05).Conclusions1. On the CIA rat model:serum IL-17level was higher than normal group; Compared with normal group,Thl7cells increased, it’s apoptosis reduced and Treg cells reduced,it’s apoptosis increased in the CIA group,There are imbalance between Th17and Treg cells 2. MTX with prednisone treatment protocals can highly reduce arthritis index, ankle swelling degree, and restrain serum IL-17level, adjust Th17/Treg balance, but it easy to cause the osteoporosis, lose weight easily.3. In MTX with chloroquin and Sulfasalazin treatment protocals, the role of improvement arthritis index, ankle swelling degree of was weaker than other two treatment groups, but it can increase Treg cells, reduce Thl7cells, to adjust Thl7/Treg balance to make RA remission. It may be the mechanism of playing the curative effect through adjust Th17/Treg balance.4. MTX with Etanercept treatment protocals can restrain serum IL-17level,and reducing the apoptosis of Treg cells, to adjust Thl7/Treg balance. Probably by adjusting the balance between Th17/Treg cells to play the role of treatment RA.5. The three treatment treatment protocals were not by promoting the apoptosis of Th17cells to adjust Th17/Treg balance, and play the role of the treatment of RA.6. Methotrexate joint yisaipu treatment protocals can adjust the balance of Th17/Treg and can restrain serum IL-17level, not easy to cause the osteoporosis, so the treatment effect of Methotrexate joint yisaipu treatment protocals is the best compared to other treatment protocals in this experiment... |
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