The Effects Of Tonifying Kidney And Supplementing Qi And Promoting Blood Recipe And The BMSCs On COL II And INOS Of Cartilage And IL-1β Of Serum In Rabbits With Knee Osteoarthritis

Objective:We research the effects of the expression of collagen II and iNOS in cartilage and the levels of IL-1βin serum of KOA rabbits by Tonifying Kidney and Supplementing Qi and Promoting Blood recipe and the BMSCs, investigate the probability of protection and mechanism, and explore a new approach of the prevention and treatment of KOA by integrated traditional and western medicine, so as to provide a new method for prevention and treatment of KOA.Methods:1. BMSCs were isolated and cultured primarily by whole bone marrow adherent method, and identified it by morphological observation, and ability of induced differentia-tion.2.30 rabbits were randomly divided into 5 groups:normal control group, model group, BMSCs group, Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group, and Combined group(Application of Tonifying Kidney and Supplementing Qi and Promoting Blood recipe and BMSCs group). The animal models of osteoarthritis were induced by papain. Intervention was given individually after two weeks, the rabbits were killed at 30 days and the articular cartilage and synovial defects were prepared for general observation, examination of the cartilage surface morphology by scanning electron microscope(SEM), pathological test by light microscope, expression of collagenⅡand iNOS by immuno-histochemical detection, and the levels of IL-1βin serum by ELISA method.Results:1. Cultured cells were identified as BMSCs by morphological observation, and ability of induced differentiation.2. General observation, surgical microscope and SEM observation:every group have different degrees of OA cartilage degeneration except for the normal control group, and the articular cartilage surface grading have statistical significance (P<0.01).3. C.Colombo integral of Histopathological examination:the model group is above the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group and the Combined group(P<0.05,P<0.01,.P<0.01). There was no significants between the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group(P>0.05). The Combined group is lower than the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group(P<0.01,P<0.05).4. The expression of typeⅡcollagen:the model group is obviously lower than the normal control group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group and the Combined group(P<0.01),and a little bit lower than the BMSCs group (P<0.05).There is no significant between the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group(P>0.05). The Combined group is above the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group(P<0.01,P<0.05).5. The expression of cartilage iNOS:there is no expression of iNOS in the the normal control group.The model group is obviously above the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group and the Combined group(P<0.01),and a little bit above the BMSCs group(P<0.05). There is no significant between the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group(P>0.05). The Combined group is lower than the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group(P<0.01,P<0.05).6. The levels of IL-1βin Serum:the model group is obviously lower than the normal control group. The model group is obviously above the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group and the Combined group(P<0.01). The Combined group is lower than the BMSCs group and the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe group(P<0.01,P<0.05).Conclusions:1. Rabbit primary BMSCs was successfully separated by whole bone marrow adherent method, and well growth of the rabbit BMSCs can be obtained after several circles of passage and purification.2. KOA rabbit model can be successfully established by papain intra-articular injection.3. BMSCs of joint cavity might induce chondrocyte differentiation, improve extracellular matrix secretion, repair of degeneration and destruction of the articular cartilage, and may play a certain role of for preventing and treating KOA.4. The Tonifying Kidney and Supplementing Qi and Promoting Blood recipe can prevent and treat KOA might through improve cartilage pathological scores, promote chondrocyte synthesize collagen II, decrease expression of articular cartilage cells iNOS, and reduce the levels of IL-1βin serum.5. The Tonifying Kidney and Supplementing Qi and Promoting Blood recipe and the BMSCs can improve cartilage pathological scores, promote chondrocyte synthesize collagenⅡ, decrease expression of articular cartilage cells iNOS, and reduce the levels of IL-1βin serum obviously. The effect is superior to that of the only the Tonifying Kidney and Supplementing Qi and Promoting Blood recipe or the BMSCs.