| Objective:In this experimental study,by constructing a model of knee osteoarthritis in rats,we explored whether Danzikang knee punch modulates chondrocytes degeneration and repair by selectively modulating the Wnt/β-catenin signaling pathway,and explored the mechanism of action of Danzikang knee punch in the treatment of knee osteoarthritis.Methods:There were 40 male rats in SD,and they were randomly divided into blank group(8 rats)and module(32 rats).The modified Hulth modeling method was applied to shape the rat KOA model.7 days after molding,rats were forced to move for 1h every day,and 2 rats were randomly selected from each group after 4 weeks to verify the modeling effect.The module-making rats were randomly divided into low-dose group,medium-dose group,high-dose group,model group,and ammonia sugar group,with 6 rats in each group and 6 in the blank group.At the beginning of week 5,the blank group and the model group were given 3mL/(kg·d)of 0.9%normal saline for gastric lavage.The high,medium and low dose groups were given 0.52g,0.26g,0.13g/(kg·d)of Danzikang knee punch for gastric gavage,respectively.The aminosugar group was given 4.27 mg/(kg·d)glucosamine sulfate capsules for gavage at a frequency of 6 times/week.Imaging after 8 weeks of intervention;Different staining methods were used to observe the pathological morphological changes of knee chondrocytes in rats.WB,RT-PCR detect protein and mRNA expression of Wnt3a,β-catenin in cartilage tissue.Results:1.General morphological observation:rough joint surface,cartilage wear,synovial thickening,osteophyte formation in the model group;Compared with the model group,the ammonia sugar group and the Danzikang knee punch group had different degrees of improvement,and the high-dose group had better effects than the ammonia sugar group and the medium and low-dose groups.2.Imaging examination:the knee joint in the model group was severely deformed,the joint space was narrowed,the cartilage thinned and disappeared,and a large number of bone spurs and osteophytes were formed.Compared with the model group,the overall improvement of rats in the knee punch group and glucosamine group was significantly higher than that in the model group,and the performance of the low-dose group and glucosamine group was roughly the same,and the overall improvement was more obvious in the high-and medium-dose groups.3.Histomorphological observation:chondrocytes in rat cartilage tissue in the model group were significantly reduced,the staining color became lighter,the tidal line was irregular and broken,and the cartilage level and structure were disordered.Glucosamine group and Danzikang knee punch had different degrees of improvement,and the performance of the high-dose group was closer to that of the blank group.4.The WB test results showed that the contents of Wnt3a and β-catenin protein decreased with the increase of the dose of Danzikang knee punch,which was lower than that of the model group(P<0.05).The high-and medium-dose groups were lower than the low-dose group(P<0.05),the low-dose group was higher than the ammonia sugar group,the medium-and high-dose groups of Wnt3a were lower than the ammonia sugar group(P<0.05),the medium-dose group of β-catenin was not significantly different from the ammonia sugar group(P>0.05),and the high-dose group decreased significantly(P<0.05).5.The RT-PCR test results showed that the relative expression of Wnt3a and β-catenin showed a downward trend with the increase of the dose of Danzikang knee punch,both lower than the model group(p<0.05),the medium and high dose groups were lower than the low dose group(P<0.05),compared with the ammonia sugar group,Wnt3a low and medium dose groups were increased,β-catenin low dose group increased,the difference between the medium dose group was not obvious,and the high dose group decreased significantly(P<0.05).Conclusion:The health status of knee cartilage in rats was negatively correlated with the Wnt/β-catenin signaling pathway.Danzikang knee punch can inhibit the expression of Wnt3a and β-catenin in the knee cartilage tissue of KOA rats,enhance the activity of joint chondrocytes in KOA rats,and promote the repair of cartilage tissue. |
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