| Objective:To observe the effects of fire needle “Dubi” and “Neixiyan” on the expression of Wnt/β-catenin signaling pathway-related proteins Wnt-4,β-catenin and MMP-13 in articular cartilage of Knee Osteoarthritis(KOA)rats.Combined with histological indexes and behavioral scores,the mechanism of fire needle “Dubi” and “Neixiyan” on KOA in rats was explored.Methods:1.Grouping and modelingThirty male SD rats were randomly divided into blank group(n(28)11)and model group(n(28)19).After 1 week of adaptive feeding,the model group was injected with 4% papanin 0.2m L on both knees on the 1st,4th and 7th day.After 2 weeks of normal feeding,3 rats were randomly selected from the blank group and model group for model identification.In the blank group,8 rats remained.The remaining 16 rats in the model group were randomly divided into a model group(n(28)8)and a fire needle group(n(28)8).2.InterveneIn the fire needle group,bilateral Dubi and Neixiyan acupoints were stimulated with fire needle,once per acupoint,once every 3 days,for a total of 4 weeks.The blank group and the model group were treated with the same grasping and fixation as the fire needle group,without treatment.3.Observe indicatorsThe weight growth rate,knee circumference growth rate,and Lequesne MG behavior score were recorded after modeling and after intervention.After the intervention,the appearance of cartilage was observed in each group.After modeling and intervention,HE staining was used to observe the pathological changes of knee joints in each group,and Mankin’s score was performed.The serum levels of IL-1β,TNF-α and MMP-13 were detected by ELISA after intervention.The protein expressions of Wnt-4,β-catenin,and MMP-13 in cartilage tissue were detected by Western Blot.Results:1.Weight gainAfter modeling,the weight growth rate of the model group was lower than that of the blank group(P<0.05).After intervention,the weight growth rate in the model group was lower than that in the blank group(P<0.05),and the weight growth rate in the fire needle group was higher than that in the model group(P<0.05).2.Growth rate of knee circumferenceAfter modeling,the growth rate of knee circumference in the model group was higher than that in the blank group(P<0.05).After intervention,the growth rate of knee circumference in the model group was lower than that in the blank group(P<0.05),and the growth rate of knee circumference in the fire needle group tended to be lower than that in the model group,but the difference was not statistically significant(P>0.05).3.Lequesne MG behavioral scoreAfter modeling,the behavioral score of the model group was higher than that of the blank group(P<0.05).After intervention,the behavior score of the model group was higher than that of the blank group(P<0.05),and the behavior score of the fire needle group was lower than that of the model group(P<0.05).4.Mankin’s cartilage histological scoreThe Mankin’s score of model identification in the modeling group was higher than that in the blank group(P<0.05),indicating that the modeling was successful.After intervention,the Mankin’s score of the model group was higher than that of the blank group(P<0.05),and the Mankin’s score of the fire needle group was lower than that of the model group(P<0.05).5.Serum levels of inflammatory factors IL-1β,TNF-α and MMP-13After intervention,the serum levels of IL-1β,TNF-α and MMP-13 in the model group were higher than those in the blank group(P<0.05),and the serum levels of IL-1β,TNF-α and MMP-13 in the fire needle group were lower than those in the model group(P<0.05).6.Relative expression of Wnt-4,β-catenin and MMP-13 proteins in cartilage tissueAfter intervention,the relative expressions of Wnt-4,β-catenin and MMP-13 proteins in the model group were higher than those in the blank group(P < 0.05),and the relative expressions of Wnt-4,β-catenin and MMP-13 proteins in the fire needle group were lower than those in the model group(P<0.05).Conclusions:1.Fire needle can reduce inflammatory response,inhibit cartilage degeneration,and improve the symptoms of KOA.2.The activation of Wnt/β-catenin signaling pathway may be involved in the occurrence and development of KOA by up-regulating the expression of MMP-13 protein.3.Fire needle “Dubi” and “Neixiyan” can reduce the inflammatory response,inhibit cartilage degeneration,and improve the symptoms of KOA in rats,which may be related to the regulation of Wnt/β-catenin signaling pathway and down-regulation of MMP-13. |
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