| Objective:To evaluate the effect of Bushen Yiganhuoxue Decoction on knee osteoarthritis in rats,and to evaluate the effect of Bushen Yiganhuoxue Recipe on articular cartilage by pathology,ELISA and immunohistochemistry.Methods:Fifty-four standard SD rats were randomly divided into blank group and 45 rats.The remaining 45 rats were injected with papain dilution model.The model group was randomly divided into model group(n = 5)The rats in the blank group were sacrificed and the rats were randomly divided into 5 groups.The rats were randomly divided into 5 groups:traditional Chinese medicine(Bushen Yigan Huoxue Decoction)high dose group,middle dose group and low dose group,model group,western medicine group(positive control group),8 rats in each group.The local changes of the model joints and the changes of the behavior of the rats in the experimental rats were observed and recorded during the drug intervention according to the pre-designed experimental method.After 4 weeks of administration,the abdominal aorta was taken and the rats were sacrificed.The cartilage tissue sections were taken by HE staining,related cytokine detection,cartilage tissue immunohistochemistry,etc.?To observe and evaluate the surface of the cartilage tissue by referring to the literature standard:After the rats were sacrificed,the model side(right posterior)knee joint was cut and the three professionals were followed according to the double-blind principle.?The pathological changes of cartilage tissue were observed by HE staining,and the scores were measured according to Mankin's score.?The contents of IL-1?and TNF-a in the serum of each group were determined by ELISA.?The expression of MMP-3 and EFEMP-1 was detected by immunohistochemistry.Results:?The performance and changes of the knee joint were made before and after the operation.Before and after the experiment,the shape and activity of the knee joint were normal.After the completion of the model,The rats in the control group showed a certain degree of modeling joint activity limitation,rat limping,local swelling and other KOA performance,after the end of the intervention of the rats in the blank group,change of the knee is not obvious,The treatment groups show a certain degree of KOA performance,but were significantly less than those in the model group.?The gross score of the knee in the normal group was lower than that in the model group and the intervention group(P<0.05),and the difference was statistically significant;The level of gross of middle?low dose group and positive control group was lower than that of model group(P<0.05),and the difference was statistically significant;Bushen Yigan Huoxue Decoction group middle dose group and positive control group were lower than those in the low dose group(P<0.05),and the difference was statistically significant;There was no significant difference in Bushen Yiganhuoxue Decoction high dose group compared with the positive control group(P>0.05)??The results of the Mankin's score were grouped after light microscopy:Mankin's scores were higher in each group than in the normal group(P<0.05),and the difference was statistically significant;The Mankin's scores of Bushen Yigan Huoxue Decoction middle dose group,low dose group and positive control group were lower than those of model group(P<0.05),and the difference was statistically significant;The high dose group,middle dose group and positive control group were lower than those of Bushen Yiganhuoxue Decoction low dose group(P<0.05),and the difference was statistically significant;There was no significant difference in Bushen Yiganhuoxue Decoction high dose group compared with the positive control group(P>0.05)??Serum IL-1?,TNF-? levels were compared between the groups:The levels of IL-1? and TNF-? in the serum of the model group and the drug treatment group were higher than those in the normal group(P<0.05),and the difference was statistically significant;The levels of IL-1? and TNF-? in the serum of the model group were higher than those in the drug treatment group(P<0.05),and the difference was statistically significant;The levels of IL-1? and TNF-? in the serum of the rats in Bushen Yiganhuoxue Decoction high dose and middle dose groups were lower than those in the low dose group(P<0.05),and the difference was statistically significant;The levels of IL-1? and TNF-a in the serum of the positive control group were lower than those of Bushen Yiganhuoxue Decoction high dose group and middle dose group(P<0.05),and the difference was statistically significant??Immunohistochemistry was used to detect the expression of MMP-3 and EFEMP-1 in cartilage tissue:The positive expression of MMP-3 and EFEMP-1 in the model group and each drug intervention group were higher than those in the blank group(P<0.05),and the difference was statistically significant;The positive expression of MMP-3 was lower in the Bushen Yiganhuoxue Decoction low?middle?high dose group and positive control group than that in the model group(P<0.05),and the difference was statistically significant;The positive expression of EFEMP-1 was higher in the Bushen Yiganhuoxue Decoction low,middle?high dose group and positive control group than that in the model group(P<0.05),and the difference was statistically significant;The positive expression of MMP-3 in the Bushen Yiganhuoxue Decoction middle?high dose group and positive control group was lower than that in Bushen Yiganhuoxue Decoction low dose group(P<0.05),and the difference was statistically significant;The positive expression of EFEMP-1 in the Bushen Yiganhuoxue Decoction middle?high dose group and positive control group was higher than that in Bushen Yiganhuoxue Decoction low dose group(P<0.05),and the difference was statistically significant;There was no significant difference in the expression of MMP-3 and EFEMP-1 between the Bushen Yiganhuoxue Decoction high dose group and the positive control group(P>0.05)?Conclusion:? The method of papain prepared by joint cavity can successfully induce rat knee osteoarthritis model,and it has a high stability.?The positive expression of MMP-3 and EFEMP-1 in model-side knee cartilage of each drug-treated group and model group was significantly higher than that in normal group,which indicated that MMP-3 and EFEMP-1 were closely related to the occurrence and development of KOA.?Bushen Yiganhuoxue Decoction can delay the degeneration of articular cartilage,thus to a certain extent,delayed the knee osteoarthritis of rats,so Bushen Yiganhuoxue Decoction on the treatment of KOA have significant clinical efficacy.?One of the mechanisms of Bushen Yiganhuoxue Decoction to delay cartilage degeneration and effectively relieve KOA disease may down-regulate and up-regulate the expression of MMP-3 and EFEMP-1 in articular cartilage.?There is no significant difference of the observation of the activity status and the histopathological observation of the rats between the Bushen Yiganhuoxue Decoction high dose group and the western medicine group(positive control group).In reducing the inflammatory factors,the positive control group was better than the Bushen Yiganhuoxue Decoction high,middle dose group.it is concluded that high doses of Bushen Yiganhuoxue Decoction can effectively treat KOA,and has the same effect with Diacerein Capsules. |
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