Study On Chondrogenic Repair Mechanism Of Papain-induced Rat Knee Osteoarthritis Model By Method Of Huashi And Bushen

ObjectiveOur previous study confirmed that Guilu Erxian Gum can inhibit cartilage degeneration in rats.The purpose of this study was to explore the mechanism of the treatment of knee osteoarthritis(KOA)by the Method of Huashi and Bushen.Methods:1.Establishment and identification of knee osteoarthritis model in SD rats The establishment of animal models of osteoarthritis by papain injection is one of the commonly used methods for the prevention and treatment of osteoarthritis.In this stu dy,Wang Zongbao's papain injection method was used frequently.According to the random number table method,48 SD rats were divided into 8 blank groups.The rest SD rats were modeled,and papain was injected into the knee joint on the 1st,3rd and 7th day respectively.The model was identified by Mankin's score and HE staining under light microscopy.2.Intervention methods: Besides the blank group,the remaining SD rats were divided into model group,Guilu Erxian gum group,Huashi Dingtong Decoction group and Guilu Erxian gum group ac cording to random number table method.The blank group and model group were fed with Normal saline,Guilu Erxian gum group was fed with Guilu Erxian gum,Huashi Dingtong Decoction group was fed with Huashi Dingtong decoction,Huashi Dington g Decoction and Guilu Erxian gum group was fed with Huashi Dingtong decoction and Guilu Erxian Gum.3.Observation of cartilage histomorphology under light microscopy Hematoxylin-eosin(HE)staining was used to observe the cartilage morphology and Mankin's score.4.The expression of p38 MAPK,type II collagen and P53 in tibial platform cartilage was detected by qPCR.5.Elisa was used to detect the expression of IL-1?,TNF-a,PGE2 and COX-2 in synovium and synovial fluid.The results:1.The results of HE staining showed that the surface tangential layer of cartilage surface in blank group was smooth and without defect,and the cells were normal in shape,uniform in arrangement and without cluster.In the model group,the tangential layer of the cartilage surface was damaged and the fissures reached the bone radiation layer,and the cells clustered.The Mankin's score of model group was significantly higher than that of blank group.The higher the score,the more serious the articular cartilage injury was.The difference was statistically significant(p < 0.05).It shows that the model of knee osteoarthritis is successful.2.HE staining results: HE staining showed that the surface articular cartilage of the blank group was smooth and flat,with uniform cell arrangement and no proliferation.In the model group,the tangential layer of cartilage surface was damaged,the fissure was deep to the radiated layer of bone,and the cells were clustered.The surface layer was destroyed,the transitional layer was intact,the cells were arranged in disorder,and there was hyperplasia.Tortoise deer erxian gluing weidingtong decoction group had smooth surface,a little damage,intact transitional layer and relatively uniform cell arrangement.3.ELISA results: The expressions of PGE,COX-2,IL-1? and TNF-a in Huashi Dingtong Decoction group were lower than those in model group(p< 0.05),and the expressions of PGE,COX-2,IL-1? and TNF-a in Huashi Dingtong Decoction and Guilu Erxian gum group were lower than those in Guilu Erxian gum group(p < 0.05).4.The expression of P38 and P53 in cartilage of Huashi Dingtong Decoction and Guilu Erxian Gum group was lower than that of Guilu Erxian Gum group(p< 0.05),and the expression of type II collagen in Huashi Dingtong Decoction and Guilu Erxian Gum group was higher than that in Guilu Erxian Gum group(p < 0.05).Conclusion:Huashi Dingtong Decoction can effectively inhibit the expression of inflammatory factors in KOA,alleviate inflammatory edema,reduce the stimulation of cartilage,reduce the apoptosis of cartilage,and then promote the repair of cartilage by Guilu Erxian Gum,and delay the process of KOA.