Experimental Study On Effects Of The Method Of Huashi And Bushen On Knee Osteoarthritis In Rats

Objective On the basis of previous studies on Guilu Erxian gum and its related prescriptions,this study discussed the mechanism of action of Huashi Dingtong decoction in the treatment of knee osteoarthritis,as well as a study on effects of the method of Huashi and Bushen(Namely Huashi Dingtong decoction adding Guilu Erxian gum)on knee osteoarthritis in rats.Methods1.Establishment and identification of knee osteoarthritis model in rats: 56 SD rats were divided into groups by random number table method: 16 SD rats were first selected for model identification and divided into Blank group and Model group,8 rats in each.The KOA model was established by injecting papain into the knee joint cavity,and the expression levels of IL-1 and IL-6 in the serum of the two groups of rats were detected.2.The experimental study on effects of the method of Huashi and Bushen on knee arthritis in rats: The remaining 40 rats were randomly divided into 5 groups: Blank group,Model group,Huashi Dingtong Decoction group(simplified Huashi group),Guilu Erxian gum group(simplified Guilu group),Huashi Dingtong Decoction and Guilu Erxian gum group(simplified Huagui group),with 8 in each group.The rats outside the Blank group were modeled,the Blank group and the Model group were given normal saline by gavage,and the other groups were given the corresponding traditional Chinese medicine by gavage,draw materials after 4 weeks of intervention.The cartilage morphology was observed by HE staining,and the expression levels of MMP-13 and MEK1/2 in cartilage were detected by immunohistochemistry.The expression of ERK1/2 in cartilage was detected by PCR.ELISA was used to detect the expression levels of NO in cartilage,NF-?B and TLR-3 in synovium,IL-1 and IL-6 in serum.Results1.Model identification results: After observation of HE staining in modeling results and Mankin scoring,it was found that: compared with the Blank group,the score of the Model group was significantly increased(P < 0.05).ELISA results showed that the expression levels of IL-1 and IL-6 in the Model group were higher than those in the Blank group(P < 0.05).This indicates that the KOA model has been successfully established.2.Results of the study on effects of the method of Huashi and Bushen on knee arthritis in rats:(1)HE staining results: The cartilage structure in the Blank group was complete,the surface morphology was normal,and the layout was close and uniform;In the Model group,there were defects in the surface layer and the transitional layer,and the cells were distributed in groups with obvious proliferation;The Huashi group was comparable to the Model group;The surface layer of Guilu group was damaged,the transitional layer was relatively complete,with hyperplasia;In the Huashi Guilu group,there was a small amount of damage in the surface layer and a complete transitional layer,with hyperplasia.(2)Immunohistochemical detection results:(1)The expression level of MMP-13 in the Guilu group was lower than that in the Model group(P < 0.05),and that in the Huashi Guilu group was lower than that in the Guilu group(P < 0.05);(2)The expression level of ERK1/2 in the Guilu group was higher than that in the Model group(P < 0.05).and that in the Huashi Guilu group was higher than that in the Guilu group(P < 0.05).(3)ELISA results:(1)The expression levels of TLR-3,NFB,IL-1,and IL-6 in the Huashi group were lower than that in the Model group(P < 0.05).The Huashi Guilu group was lower than the Guilu group(P < 0.05);(2)The expression of NO in the Guilu group was lower than that in the Model group(P < 0.05).The Huashi Guilu group was lower than the Guilu group(P < 0.05).(4)q PCR results:the expression level of ERK1/2 in Guilu group was higher than that in Model group(P < 0.05),and the expression level of ERK1/2 in Huashi Guilu group was higher than that in Guilu group(P < 0.05).Conclusion1.By reducing the inflammatory edema and local swelling of the articular cartilage and synovium of the rats,Huashi Dingtong decoction can enhance the original curative effect of Guilu Erxian gum decoction.2.Use the method of Huashi and Bushen,the effect of the Huashi Guilu group of chondrocytes apoptosis is better than the Guilu group.This was related to the fact that the combination of the Huashi Dingtong decoction with the Guilu Erxian gum decoction could reduce the expression of NF-B,TLR-3,IL-1,and IL-6 to inhibit the activation of the MEK/ERK signaling pathway,thereby reducing the expression of NO and MMP-13.