Effect Of Fuyuan Capsule On Cartilage Matrix And Nitric Oxide,Inducible Nitric Oxide Synthase,Prostaglandin E2 Of Inflammatory Mediators In Knee Osteoarthritis

Objective:1 . To investigate the effect of Fuyuan capsule ( FYC ) on histomorphology of articular cartilage and expression of cartilage matrix in experimental knee osteoarthritis(KOA) as well as to further explore the mechanism how does it perform in protection of articular cartilage against OA.2.To investigate the effect of FYC on the expression of inducible nitric oxide synthase (iNOS),nitric oxide (NO) and prostaglandin E₂(PGE₂) of inflammatory mediators in experimental KOA, and explore the anti-inflammatory action of FYC on OA.Methods:Sixty male Sprague-Dawley(SD) rats were randomly divided into 6 groups: normal group, model group, tetracycline group, FYC low-dose group, FYC middle-dose group and FYC high-dose group. OA model was induced by improved Hulth's method. Two weeks later, the treated groups were respectively given different dose of drugs administered once a day: R ats in FYC groups were given Fuyuan capsule 371.65 mg·kg^-1·d^-1,743.30 mg·kg^-1·d^-1 and 2229.90 mg·kg^-1·d^-1, and rats in tetracycline group was given tetracycline 50 mg·kg^-1·d^-1. 12 weeks later after administering, obtain the blood from Abdominal aorta and cut the condylus medialis femoris from the right knee joint of Rats. Histomorphology of articular cartilage was observed by general and HE staining. The levels of proteoglycan and collagen in cartilage were tested by staining with toluidine blue and Masson respectively. Moreover, the expression of collagen type II,collagen type I and iNOS in articular cartilage was detected by immunohistochemistry. Meanwhile contents of NO and PGE₂ in serum were also tested by nitrate reductase and enzyme-linked immunosorbent assay respectively.Results:1.Effect of FYC on histomorphology of articular cartilage in KOA The surface of cartilage in normal group was smooth and glossy,but that in model group was rough. Chondrocytes of light microscopein in normal group arrayed regularly, and cartilage matrix was dyed uniformly. But chondrocytes in model group were very indiscriminate and its numbers were decreased. Also, chondrocytes in FYC groups showed tidily and numbers were increased compared with model group. The general scores and Mankin's scores in model group were higher than normal group(P<0.05), and that in FYC groups were lower than model group(P<0.05).2.Effect of FYC on cartilage matrix in KOA(1) Contents of proteoglycan and collagen: cartilage staining with toluidin blue and Masson in normal group showed uniformly, however that in model group showed unevennessly. While FYC groups were well-staining than model group. Compared with normal group, the contents of proteoglycans and collagen in model group were minorer remarkably(P<0.01), and those in FYC groups were more than model group(P<0.05).(2) Expression of collagen type II and collagen type I: In normal group, we could view the expression of collagen type II in cartilage of full-thickness, but could not view the expression of collagen type I in cartilage.Compared with normal group, the expression of collagen type II in model group was lower remarkably(P<0.01), but that of collagen type I was higher(P<0.01). FYC groups show higher expression level of collagen type II(P<0.05), and lower expression level of collagen type I (P<0.05) compare to model group.3.Effect of FYC on expression of iNOS in cartilage and NO,PGE₂ in serum in KOA(1) Expression of iNOS: There was very little expression of iNOS in normal group artilage cartilage. But in model group, the expression of iNOS was viewed in cartilage of full-thickness, and was higher remarkably than normal group (P<0.01). The expression of iNOS in FYC groups were lower than model group (P<0.05).(2) Content of NO: The level of NO in model group was higher remarkably than normal group (P<0.01), and FYC groups were lower than model group (P<0.05).(3) Content of PGE₂: The level of PGE₂ in model group was higher remarkably than normal group (P<0.01), and FYC groups were lower remarkably than model group (P<0.01) .Conclution:1. We can view typical manifest of OA after the model was induced for 12weeks, which indicate that OA model was successed. FYC could alleviate pathological injuries of articular cartilage in KOA, which indicate that FYC have a protective role in cartilage.2. FYC can enhance the expression of proteoglycans and collagen (especially for collagen type II), reduce the expression of collagen type I as well, which would maintain the normal collagen phenotype and protect articular cartilage.3. FYC can reduce the expression of iNOS in cartilage and levels of NO and PGE₂ in serum in osteoarthritis, which would have some prevent effect in alleviating arthritis.