| Objective:1.Objective To observe the physiological and pathological effects of regulating knee on knee osteoarthritis(KOA),interleukin-1 beta(IL-1β)and tumor necrosis factor-alpha(tumor necrosis factor-alpha)in rabbits.TNF-α).2.Based on Wnt/β-catenin signaling pathway,the protective mechanism of knee-regulating method on KOA rabbit cartilage was investigatedMethods:New Zealand rabbits were randomly divided into normal group,model group,manipulation group and drug group,8 in each group.KOA model was replicated by intra-articular injection of papain solution.No intervention was given to the normal group and the model group.Manipulative group was treated with knee adjustment method every day,10 min each time,while the drug group was treated with celecoxib solution by Gavage every day.After continuous treatment for 21 days,the samples were collected and the levels of IL-1β and TNF-α in serum were measured by Elisa,and the histological morphology of knee articular cartilage was observed by H&E stain method and saffron-green method The expression levels of IL-1β and TNF-α in cartilage were observed by immunohistochemistry,and their integrated optical density(IOD)was analyzed The m RNA and protein levels of WNT3Α,β-catenin and matrix metalloproteinase metalloproteinase-13(MMP-13)in cartilage were determined by real-time quantitative PCR and Western blot,respectively.Results:1.Serum ELISA test results showed that compared with normal group,serum levels of IL-1β and TNF-α in model group were significantly increased,and the difference was statistically significant(P<0.05).Compared with model group,the serum levels of IL-1β and TNF-α in manipulation group and drug group were significantly decreased,with statistical significance(P<0.05).The serum levels of IL-1β and TNF-α in drug group were significantly lower than those in manipulation group,and the difference was statistically significant(P<0.05).2.The HE stain method and the saffron-green method showed that the cartilage of the knee joint in the normal group was intact,the surface of the cartilage was smooth,there was no exfoliation on the surface,and there was no abnormal change in the cartilage layer and nucleus.In the model group,the structure of cartilage was disordered,the surface of cartilage was rough,the surface layer of cartilage was denuded,the thickness of cartilage layer was thin,and the number of chondrocytes was decreased.The above pathological changes in the manipulation group and the drug group were thicker than those in the model group,cartilage denudation was less,and nuclear degeneration was milder,and the pathological changes in the drug group were improved more obviously than those in the manipulation group.3.Compared with the normal group,the IOD of the model group was significantly increased(P<0.05),and the IOD of the model group was significantly higher than that of the normal group(P<0.05),the positive expression of cartilage in manipulation group and drug group decreased significantly(P<0.05),and the positive expression of cartilage in drug group decreased significantly(P<0.05).4.Real-time quantitative PCR detection of Wnt3α,β-catenin and MMP-13 m RNA showed that compared with normal group,the m RNA expression levels of Wnt3α,β-catenin and MMP-13 in model group were significantly increased,and the difference was statistically significant(P<0.05).Compared with model group,m RNA expression levels of Wnt3α,β-catenin and MMP-13 in cartilage tissues of manipulation and drug groups were significantly decreased,and the difference was statistically significant(P<0.05).The expression level of drug group was significantly lower than that of manipulation group,and the difference was statistically significant(P<0.05).5.Western blot analysis of wnt3α,β-catenin and MMP-13 protein in cartilage showed that the expression levels of wnt3α,β-catenin and MMP-13 protein in the model group were significantly higher than those in the normal group,compared with the model group,the expression levels of Wnt3α,β-catenin and MMP-13 in the cartilage of the manipulation group and drug group were significantly decreased(P<0.05)The expression level in the drug group was significantly lower than that in the manipulation group(P<0.05).Conclusions:1.Pathological changes were observed in rabbit cartilage osteoarthritis of KOA in the model group,and the expression levels of IL-1β and TNF-α in serum were increased,suggesting that the classic KOA model was successfully established,and the inflammatory mechanism was involved in the progression of KOA.2.After the intervention of knee-regulating therapy and celecoxib,the pathological state and inflammatory reaction of cartilage in KOA rabbits were improved,which indicated that knee-regulating therapy and celecoxib had good therapeutic effect,and the therapeutic effect of drugs was better than that of manipulation.3.Regulating knee can inhibit the inflammatory response and improve the pathological status of knee cartilage,and its mechanism may be related to blocking the activation of Wnt/β-catenin signaling pathway. |
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