Experimental Study On Cervus And Cucumis Polypeptide Against Articular Cartilage Degeneration Of The Knee OA Models In Rabbit By Intraarticular Injection

Objective:To study effect and protective mechanisms of Cervus and Cucumis Polypeptide against articular cartilage degeneration of the knee OA models in Rabbit by intraarticular injection;Methods:Fifty health adult New Zealand rabbits for experiments were randly divided into 10 normal rabbits as the normal group and 40 models rabbits whose unilateral hind joints were fixed by plaster cast. After the OA modeling, they were avergely divided into 4 groups (model group, HA group, Cervus and Cucumis Polypeptide InjectionⅠgroup,Ⅱgroup) in random. Except the normal group, the other four groups were injected the corresponding drug by intraarticular. After 5 weeks, rabbits were sacrificed by air embolism.(1) Cut apply adequate amount of articular cartilage and made ordinary optical microscope slides and transmission electron microscope slides, to observe ultrastructural articular cartilage pathology. According to the Mankin articular cartilage of trypan-blue and histological scoring system for the pathological changes of the cartilage tissue and compare half grading the differences between groups. (2) we extracted joint fluid, measured,the levels on NO and IL-1βand observed synovial inflammation in each group.(3) we observed the mRNA expressions of MMP-3,MMP-13 and TIMP-1 in knee cartilage of OA model rabbit by RT-PCR;(4) We cut the right amount of articular cartilage of rabbit femoral condyle, and respectively measured the level of GAG and positive stain area of Collagen TypeⅡin each group By IHC, and compared the differences between groups.Results:(1) In macroscopic view, articular cartilage of the HA groups,Cervus and Cucumis Polypeptide InjectionⅠgroup were the basic same in pathological changes:Surface was mostly yellow, patchy or diffuse slightly rough, hadn't the obvious crack or defect; In Cucumis Polypeptide Injection II group articular cartilage surface was light yellow and smooth; Light and electron microscopic observation found that after medical treatment the form, structure, arrangement, etc of the articular cartilage cells of Cervus and Cucumis Polypeptide InjectionⅡgroup were better than HA group, Cervus and Cucumis Polypeptide Injection I group.(2) Before treatment the levels on NO and IL-1βof each group no statistically significant difference(P>0.05). After treatment there weren't significant differences between Cervus and Cucumis Polypeptide Injection I group and HA group(P>0.05); While the levels on NO and IL-1βof Cervus and Cucumis Polypeptide Injection II group were lower than others, and showed significant difference (P<0.05). Synovium of the three groups—the HA group,Cervus and Cucumis Polypeptide InjectionⅠ,Ⅱgroup—no or mild redness, swelling, hyperplasia, and the synovitis significantly reduced compared with model group.(3) In the normal group, the mRNA expression rates of MMP-3, MMP-13 were relatively low, and TIMP-1was relatively high; while in the model group, the mRNA expression rates of MMP-3, MMP-13 significantly increased and TIMP-1was significantly low; After treatment, the mRNA expression rates significantly decreased in the three treatment groups. Between Cervus and Cucumis Polypeptide Injection I group and the HA group, no statistically significant difference (P>0.05), while Between Cervus and Cucumis Polypeptide InjectionⅡgroup and the HA group, showed significant difference after treatment (P<0.05).(4) GAG content results in each group:①The GAG content of cartilage matrix was no significant difference between the model group and the HA group (P>0.05);②Cervus and Cucumis Polypeptide InjectionⅠ,Ⅱgroup and the HA group, model group were compared, the GAG content of cartilage matrix of Cervus and Cucumis Polypeptide Injection groups were significantly different (P<0.01);③The GAG content of cartilage matrix was significant difference between Cervus and Cucumis Polypeptide InjectionⅠandⅡgroup (P>0.05). Results of positive stain area of Collagen TypeⅡin each group:①The positive stain area of Collagen TypeⅡwas no significant difference between the model group and the HA group (P>0.05);②Cervus and Cucumis Polypeptide InjectionⅠ,Ⅱgroup and the HA group, model group were compared, The positive stain area of Collagen TypeⅡof Cervus and Cucumis Polypeptide Injection groups were significantly different (P<0.01);③The positive stain area of Collagen Type II was significant difference between Cervus and Cucumis Polypeptide Injection I and II group (P>0.05).Conclusions:(1) Cervus and Cucumis Polypeptide Injection could significantly reduced the levels on NO and IL-1βin the articular cavity of the knee OA model in rabbit, and could inhibit synovitis disease. (2) Cervus and Cucumis Polypeptide Injection could inhibit the mRNA expressions of MMP-3, MMP-13 in Knee Cartilage of OA Model Rabbit by Intraarticular Injection, and increase the mRNA expressions of TIMP-1,and delay the degeneration of articular cartilage.(3) Cervus and Cucumis Polypeptide could not only improve the GAG content of cartilage matrix, and promote the synthesis of collagen typeⅡ, and the results had a certain relationship with the dose.(4) Cervus and Cucumis Polypeptide could promote extracellular matrix synthesis, and degradation of newly synthesized matrix was significantly inhibited.By the naked eye, light microscopy, and electron microscopy, results showed that:Under the common function of various factors in the above, Cervus and Cucumis Polypeptide could promote the synthesis of collagen typeⅡ, increase GAG content, inhibit degradation of ECM, reduce or control inflammation of KOA, thus maintain and keep the articular cartilage normal structure and function, prevent the process of degenerative diseases of rabbit KOA model.