Study Of Relations Between Inflammation Network Factors And The Prethrombotic State In Rats With CIA And Its Intervention By Traditional Chinese Medicine

Objective:In order to evaluate the effects of inflammation network factors on relationship between the pathopoiesis of immune inflammatory vasculopathy in rheumatoid arthritis (RA) and prethrombotic state, a model of rats with collagen typeâ…¡-induced arthritis (CIA) was established to explore the relationship between inflammation network and the prethrombotic state in CIA rats, and to test the intervention of traditional Chinese medicine, salvia miltiorrhiza and berberine.Methods:A model of rats with collagen-induced arthritis (CIA) was established by intradermal injection with collagen typeâ…¡emulsified with complete Freund's Adjuvant (CFA). Next day, rats with CIA in the administration groups which were divided into 5 groups were administrated by intragastric administration, while the rats with CIA in model group were daily administration of 1.5ml of distilled water. And the normal control group drink freely. The mean arthritis index (AI) of rats in each group was scored every week. Blood samples were collected from the heart of the rats on the 35th day, and the sera were immediately separated by centrifugation. IL-6 and D-dimer concentrations in sera of rats from each group were determined with the enzyme-linked immunosorbent assay (ELISA), and the level of MMP-9 in sera of rats from each group was evaluated by Western Blotting, and the relative concentration of MMP-9 was also performed. Knee joints, resected from the sacrificed rats were estimated in routine knee pathology, and the expression of CD40 in the synovium was detected using immunohistochemistry assay.Results:1. To copy the RA model successfully, the symptoms of arthritis in rats, such as a clear local and continuing joint swelling, joint motion handicap to clinical manifestations, were induced using collagen typeâ…¡emulsified with complete Freund's Adjuvant, and that showed the model of RA successfully. Histopathologically, synovial thickening, and a large number of inflammatory cells diffused or locally infiltrated, and fibroplasias were observed.2. The levels of IL-6, MMP-9 and D-D in sera of rats with CIA were much higher than that in control group (P<0.05), and the expression of CD40 in the synovium was also raised obviously (P<0.05). Compared with the model group, they decreased in varying degrees in administration groups, while the arthritis index score dropped correspondingly.3. Compared with that in model group and in berberine group, the levels of IL-6, MMP-9 and D-D in sera and expression of CD40 in synovium of rats in administration groups were significantly dropped (P<0.05). Correspondingly, the symptom of arthritis relieved and the arthritis index score changed. Moreover, the above-mentioned items in salvia miltiorrhiza plus MTX group were much lower than that in MTX alone group, salvia miltiorrhiza alone group, and berberine plus MTX group (P<0.05).4. Compared with that in model group, the levels of IL-6, MMP-9 and D-D in sera and the expression of CD40 in the synovium of the rats in berberine plus MTX group dropped dramatically (P<0.05), while in berberine alone group, they had a certain impact on the above-mentioned items, but there were not statistically significant (P>0.05).Conclusions:1. The increased release of inflammation network factors in the pathogenesis of rat with CIA is involved in the process of immune arthritis and synovium pathological changes in RA. And it is also the pathological basis of promoting the prethrombotic state or local thrombogenesis of acra in RA.2. The therapeutic effect of combined salvia miltiorrhiza and MTX was significantly better than that of single administration. The results presented here demonstrated that treatment with salvia miltiorrhiza plus MTX led to synergistic anti-immune and anti-inflammatory effects, decreased the levels of immune inflammation network factors, and improved the prethrombotic state which caused by immune inflammation.3. To a certain degree, combination of berberine and MTX decreased the expression or release of the inflammation network factors in rats with CIA, however, it's effectiveness is still inferior to treatment with salvia miltiorrhiza plus MTX. It is not so effective to use berberine alone.