The Effect Of Leflunomide And Breviscapine On IL-1,IL-6,TNF-α And MMP3/TIMP1 In The Animal Model Of Rheumatoid Arthritis

Objective: Rheumatoid arthritis(RA)is a systemic disease of unknown etiology that is characterized by chronic polyarticular joint inflammation which lead to destruction of cartilage and bone. Histopathologic features of RA encompass infiltration by macrophages and T cells synovial lining hyperplasia,neoangiogenesis,pannus formation and destruction of cartilage and bone. Study recently show that there is a correlation between cytokings IL-1,IL-6,TNF-α, MMP3/TIMP1 and bone damage. Update,inhibiting joint destruction of cartilage is the major theapy. We found Breviscapine had a good effect in treating RA .The aim of this study is to investigate the therapeutic mechanism of Bre on cytokine IL-1,IL-6,TNF-αand MMP3/TIMP1 in the animal model of rheumatoid arthritis .Methods: (1)Fifty male Wistar rats were randomly divided into 5 groups:the normal group,the model group,the leflunomide group,the Bre group,the lef and bre group.Except for the rats of normal control group,the others were intracutaneously injected with 0.1ml Freund,s complete adjuvant in the right hind limb. (2) The medicines were given from the 18th day to the 32th day once a day to observe its effect on multiple arthritis. AA was assessed :by measuring arthritic index,paw volume,general status and body weight. (3)The levels of IL-1,IL-6,TNF-αwere measured with radioimmunoassay. (4) Under high microscope,we observe the pathological change of ankle joint and record the remark,in order to study the effect on cartilage destruction in AA in rat. (5) Immunohistochemistry was used to detected the expression of MMP3/TIMP1in synovial tissue. The intensity of MMP3/TIMP1 in synovial tissue was analyzed by computerized image system. (6)There was positive correlation between IL-1 and MMP3 ,between pathological change and MMP3.Results 1 bre, lef and combined group could markedly reduce the degree of rat pedal swelling,improve general status,increment the body weight as compared to the model group(P＜0.05).There was difference between model group and combined group(P＜0.05).The increment body weight of combined group had no difference as compared to the control group.2 There was no significantly difference of arthtitis indexs and paw swelling in every group after modeled group .After treatment,every group could reduce the arthritis indexs and paw swelling.there was difference as model group.The effect of combined group was superior to the lef group or bre group.3 The levels of IL-1,IL-6,TNF-αin model group was higher than control group.After treatment,the serum IL-1,IL-6,TNF-αdecreased significantly in the bre group as compared to that in the model group. there was significantly difference(P＜0.05).the levels of IL-1,IL-6,TNF-αof combined group were lower than bre and lef groups(P＜0.05).4 The pathologicals results showed that there was a lot of macrophages and T cells in synovial tissue. There was pannus formatted in synovial tissue. The remarks of model group was higher than that in control group.5 The Immunohistochemistry showed that the expression of MMP3 in model group was higher than control group.Bre could decrease the expression of MMP3.there was difference between bre group and model group. The expression of MMP3 in combined group was most lowest in test group. The result of TIMP1 was contrast to the expression of MMP3. 6 The correlation between serum IL-1,pathological remark and MMP3 was positive significantly(P＜0.01).The result showed that IL-1 played an important role in joint destruction. In the other hand, the result also showed that MMP3 was an important factor in cartilage and bone.Conclusion: 1 The result showed that bre had a defined effect in treating animal model of rheumatoid arthritis. It could reduced the paw swelling and arthritis indexs; It could decrease the levels of serum IL-1,IL-6,TNF-α; Moreover, Bre could decrease effectively the expression of MMP3 in synovial tissue. To sum up, Bre could relieve AA symptoms and suppressed cartilage destruction.