| Objective:To elucidate the mechanism of the pathogenesis of rheumatoidarthritis(RA)by exploring the expression of RhoA in synovium of collagen-induced arthritis(CIA) rat, observe the protection of BZXD on cartilage injury in synovium of CIA rats, and provide experimental evidence for BZXD treating RA.Methods:1,The experiment arthritis rat model was established by injected subcutaneous of bovine II collagen(BIIC)and complete Frennds adjuvant(CFA). The CIA rats were divided into 2 groups randomly, the Bizhongxiao decoction treatment group(BZXD group, n=15)drenched with BZXD and the CIA model group(Model group, n=15) drenched with the same dose of distilled water. Normal control group(Normal group, n=15) drank water freely.2,General condition, weight,arthritis index score. degree of paw swelling and pathology was observed in 3 groups. The arthritis index(AI)of the rats was examined in CIA group and BZXD group on 10th day,15th day,20th day,25th day,30th day,35th day,40th day and 45th day respectively. The weight and ankle thickness of rats were measured in three groups every week. The pathomorphology of ankle was observed on 25th day,35th day,45th day. 3,RhoA expression of synovium at different stages from the model, BZXD and normal group were tested by immunohistochemisty,and analyzed by Computer image analysis system of gray scale. Diversity comparation was assayed by SPSS 11.0 software.Results:1. Copying CIA model:CIA model could be copied by BIIC and CFA with the incidence of 90.0% approximately.(1)General condition:poor appetite was shown in rats of model group, more weight was losen than BZXD group(P<0.05)as a result. Laterly,symptoms as dry and withering hair were manifested, whereas such symptoms were not found in BZXD group. The weight increment in model group was lower than that in normal group in the 3rd w, the 4th W, the 5th W, the 6th w(p<0.05), respectively,without significant difference between them in the 1st w. the 2nd w. The weight increment in BZXD group was higher than that in model group in the 4th w, the 5th w,the 6th w,with significant difference between them (P<0.05), no significant difference among them at other times.(2)Arthritis index score evaluation:Arthritis of CIA rats was found on 6-8 days after injection. Arthritis index score was found increased in model group and BZXD group on 10 days, reaching a peak on 25 days and turning decreasing on 30 days in model group and BZXD group. But there was no manifest difference in arthritis index score between model group and BZXD group before 20 days(P>0.05).2 weeks after treatment, arthritis index score in BZXD group was found decreasing. AI in BZXD group was lower than that in model group on 20d,25d,30d,35d,40d, 45d(P<0.05), respectively.(3)Swelling degrees evaluation in joints of claws and feet:The ankle thickness of foot in model group and BZXD group was thicker than that in normal group in the 1stw, (P<0.05), respectively. The ankle thickness of foot in BZXD group was thinner than that in model group after the 5th w, (P<0.05), respectively.(4)Observation of pathology:Rat's synovial tissues was not manifested by obvious newborm blood vessel and infiltrating inflammatory cells in normal group. While CIA model group on 30 days was manifested by angiogenesis and increased synovial membrane. With aggravation of symptoms, newbom blood vessels were increasing. Synovial tissues a few newborn blood vessels and infiltrating inflammatory cells as well as slightly increased synovial membrane was showed in BZXD group. On following 45 days, the newborn blood vessels decreased, with relieving inflammation and thinner synovial membrane. Pathologic injures could be ameliorated by BZXD.2.The test of RhoA expression in synoviumThe immunohistochemisty analysis showed that, the gray scale of RhoA in model group was lower than that in normal group, and 45days lower than 30days, (P<0.05), respectively. The gray scale of RhoA in BZXD group were obviously higher than that in model group, and 45days higher than 30days in BZXD group, (P<0.05), respectively.Conclusions:1. Subcutaneous injection of BIICand CFA imitates CIA model successfully, which can be used in as research in RA.2. During the process of CIA,RhoA expression was significantly enhanced,and with arthritis becoming more severe, RhoA expression edged up, prompted that RhoA may be involved in the process of RA synovium and cartilage disease.3.BiZhongXiao Decoction can down regulate RhoA expression in synovial tissue of Collagen Induced Arthritis rat, which is one of the probable mechanism of treating RA.
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