| Background:Through the study of the pathogenesis and various treatment methods of KOA,the basic principle of "urgent loosening tendons and strengthening bones,slow tonifying kidney and strengthening bones" was formed.therefore,the experimental optimization of knee joint shaking exercise rapid pain relief,Bushen Huoxue recipe for the formation of a permanent cure,this scheme can not only quickly relieve pain,but also prevent repeated attacks of pain symptoms in KOA,so as to delay the progress of KOA.Purpose:The purpose of this study is to explore the effect of Bushen Huoxue recipe + traction method on cartilage inflammation of knee osteoarthritis by observing the changes of morphological degeneration and damage of knee cartilage in rats,and through the changes of NLRP3 inflammatory corpuscle signal pathway and downstream related factors in cartilage tissue,so as to provide experimental basis for the prevention and treatment of knee osteoarthritis.Method:Fifty rats were fed adaptively for one week and randomly divided into blank control group(n = 10)and intervention group(n = 40).The rats in the intervention group were induced by sodium iodoacetate method.On the 1st,4th and 7th day,0.05ml(40mg/ml)was injected into both knee joints to establish KOA rat model,while the blank control group(both knees)was injected with the same amount of normal saline.In our early KOA rat experiment,it has been verified by double knee 2min,double knee 4min and double knee 6min that the effect of double knee 4min is the best.After the establishment of the model,40 rats in the intervention group were randomly divided into model group,knee joint shaking group(double knee 4min),Bushen Huoxue recipe group(traditional Chinese medicine group),knee joint shaking and Bushen Huoxue recipe combined group(Bushen Huoxue recipe intragastric administration + double knee 4min),with 10 rats in each group.The blank control group was given the same volume of normal saline once a day for 8weeks.The model group was given the same volume of normal saline once a day for 8weeks.Bushen Huoxue recipe group was treated with Bushen Huoxue recipe 3.2ml/ only once a day for 8 weeks.The knee joint traction group was given knee joint traction every2 days,with 4min of both knees every time for 8 weeks.The combined group was treated with Bushen Huoxue recipe(3.2ml)by intragastric administration once a day + knee joint shaking method was performed every 2 days,4min of both knees every time for 8 weeks.After the rats were killed,the experimental results were observed by gross articular cartilage section staining,Mankin’s score,NLRP3 and other key inflammatory factor protein pathway and m RNA expression.The experimental results were statistically analyzed by SPSS22.0 statistical software.Results:1.Gross observation of articular cartilage in rats In the blank group,the surface of articular cartilage was smooth and complete,there was no local cartilage destruction,no cracks,a small amount of transparent synovial fluid,normal synovial tissue,no inflammatory and pathological changes of joints.Model group:articular cartilage surface uneven,articular cartilage thinning,local wear and destruction,increased articular fluid,viscous follicular changes,yellowish,synovium hypertrophy,the whole joint showed inflammatory changes.The surface of articular cartilage in the traction group was more uneven than that in the blank group,with a small amount of crack destruction,less synovial fluid and no obvious pathological changes in synovium.The surface of articular cartilage in the traditional Chinese medicine group was more uneven than that in the blank group,with local wear and tear,increased articular fluid and cracks.In the combination group,the joint surface was smooth and complete,light blue,there was no obvious cartilage destruction,more articular fluid,some of them showed sticky follicular changes,the color was lighter,and the synovium had no obvious pathological changes.2.Results of staining and Mankin’s score of gross articular cartilage sections in ratsCompared with the model group,the articular surface of the articular cartilage section combination group was smooth and complete,there was no obvious cartilage destruction,and there were no obvious pathological changes of synovium.There were significant differences in Mankin’s score between blank group and other four groups,between combination group and traditional Chinese medicine group,and between traditional Chinese medicine group and draft group(P < 0.05).3.Protein pathway and m RNA expression of NLRP3 and other key inflammatory factors3.1 Results of Western blot detection of knee joint cartilage in SD ratsThe expression of NLRP3,Caspase-1 and IL-1β protein in the cartilage tissue of the traction group was much lower than that of the traditional Chinese medicine group,and the expression of NLRP3,Caapase-1 and IL-1β protein in the traditional Chinese medicine group was significantly lower than that of the model group.The protein expression levels of NLRP3,Caspase-1 and IL-1β in the combination group were significantly lower than those in the model group(P < 0.001).3.2 Expression of related m RNA in cartilage of SD ratsCompared with the model group,the m RNA expression levels of IL-1,caspase-1 and NLRP3 in the cartilage tissue in the traction group were significantly lower than those in the model group,and the m RNA expression levels of IL-1,caspase-1 and NLRP3 in the traditional Chinese medicine group were significantly lower than those in the model group.The m RNA expression of IL-1,caspase-1 and NLRP3 in cartilage tissue in the combination group was significantly lower than that in the model group.Conclusion:Shaking method and Bushen Huoxue recipe intervention method can effectively protect and repair the morphological injury of cartilage in KOA model of SD rats,and the effect of both is the best. |
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