| Objective:To establish the rat model of adjuvant-induced Arthritis, to observe the therapeutic effect of electroapuncture(EA)on Zusanli(ST36),Guanyuan(CV4), A-shi point of adjuvant arthritists(AA) rats' c-IAP1 ? c-IAP2 of gambrel synovial tissue apoptosis inhibition proteins, in order to investigate the possible mechanism of electroapuncture in treating rheumatoid arthritis(RA).Methods: A total of 60 male SD rats were randomized into control, model, MTX and EA, 15 for each group. Adjuvant arthritis model(AA) was established in male SD rats with subcutaneouly injection of Mtb(0.1m L) in mineraloil at the base of the tail of animals(except for the rats of the nomal control group). Intervention treatment was conducted in second days,nomal group and model group received no treatment, while rats in the MTX group was given methotrexate to fill the stomach; EA group rats was treated electroapunctring everyday once,1course for 7 times, 21 times(3 courses) in all. During the course of the experiment, to observe the general situation, body weight, the degree of paw edema and arthritis index(AI) every three days. At he end of the experiment, to observe the synovial of ankle joint pathological changes by light microscopy. Test the positive expressions of inhibitor of apoptosis proteins c-IAP1 and c-IAP2 using immunohistochemistry. And the expressions of apoptosis inhibitor m RNA c IAP1 and m RNA c-IAP2 were detected by reverse transcription polymerase chain reaction(RT-PCR).Results:(1) Effect of body weight, paw edema and arthritis index of rats: Compared with the nomal control group, the model group had decreased body weight,increased degree of paw edema and arthritis index, the difference was statistically significant(P<0.01); Between the MTX and the nomal control group, the difference was no significant(P>0.05). Compared with the model group, The paw edema, arthritis index both decreased in MTX and EA group,and the difference was statistically significant(P<0.01, P<0.05). Compared with the MTXgroup,higher degree of paw edema and arthritis index in EA group,and the difference was statistically significant(P<0.05).(2)Histological changes of rat's synovial of ankle joint in each group:In the nomal control group, rats synovial tissue had no hyperplasia, arranged in neat rows and showed a few small blood vessels; Model group rats ankle joint synovial tissue showed obvious synovial hyperplasia,cells infiltration, pannus formation and cartilage destruction; EA group rats synovial tissue showed hyperplasia,as thick as 3-5 layer and a few inflammatory infiltration,no obvious pannus formation and cartilage destruction. MTX group rats synovial tissue hyperplasia is not obvious, few cells infiltration,which were close to normal.(3) Effect of immunohistochemistry on the ankle joint in rats: Compared with the nomal control group, the c-IAP1?c-IAP2 positive expressions of synovial tissue were increased in model group,the difference was statistically significant(P<0.01); Between the MTX group and the nomal group, the difference was no significant(P>0.05). Compared with the model group, the c-IAP1?c-IAP2 positive expressions of synovial tissue of the EA and MTX group rats were decreased(P<0.05),but the positive expression of EA group was higher than that of MTX group(P<0.05).(4) RT-PCR results of synovial tissue of rats in each group:Compared with the nomal control group, the expression of model group in synovial tissue apoptosis inhibition of protein c-IAP1?c-IAP2 m RNA were higher(P<0.01). Between the MTX group and the nomal group,the difference was no significant(P>0.05).Compared with the model group, EA and MTX group rats apoptosis inhibitionof protein c-IAP1?c-IAP2 m RNA expression were lower than the model group,and the difference was statistically significant(P<0.05).Compared with the MTX group, higher expression of protein c-IAP1 ? c-IAP2 m RNA in EA group,and the difference was statistically significant(P<0.01).Conclusion:(1) Both electroapuncture and methotrexate can reduce AA rats paw edema, arthritis index,ankle synovial tissue hyperplasia,synovial inflammatory cells infiltration, pannus formation and cartilage destruction.And the effect of methotrexate in this area is close to the normallevel, but electroapuncture is less effective compare with methotrexate. It indicated that although the effect of electroacupuncture is effective,but it can not reach the normal level.(2)The electroapuncture and methotrexate can reduce synovial tissue apoptosis inhibition of protein c-IAP1?c-IAP2 m RNA expression, its mechanism may be related to the inhibition of c-IAP2, c-IAP1 m RNA expression, inducing apoptosis of synovial cells.(3) Methotrexate as the internationally recognized drug in treatment of rheumatoid arthritis,and its effect is better than the electroapuncture,the differences indicate that the action way and mechanism of the treatment of electroapuncture and methotrexate is not the same, A further studyis needed indeed; on the other hand, considering the study of electroapuncture cycle is not long enough and shorter course of treatment. |
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