Study On The Efficacy And Mechanism Of Jiawei Duhuojijie Mixture In Promoting Cartilage Repair Of Knee Osteoarthritis

Objective: 1 Animal experiments:To establish the animal model of Knee Osteoarthritis(KOA)in combination with traditional Chinese medicine(TCM)pathogenesis of "deficiency,blood Stasis and toxin",and to discuss the effects and mechanism of Jiawei Duhuo Jisheng Mixture on promoting the repair of KOA cartilage.2 Clinical experiments: A randomized controlled trial was used to verify the effects of Jiawei Duhuo Jisheng Mixture in the treatment of KOA,and to evaluate the safety of its clinical application.Methods: 1 Animal experiments: A method of plaster fixation,hydrocortisone feeding and 4? ice water stimulation was used to establish the KOA model.Then,rabbits were randomly assigned to Groups A(Control),B(TCM),C(positive control)and D(Model).Rabbits of group A and D were fed with normal saline(NS),rabbits of group B were fed with Jiawei Duhuo Jisheng Mixture(2.26 g/kg)while rabbits of group C were fed with diacerein capsules(2.725 mg/kg)twice a day.Six weeks later,rabbits were sacrificed and joint fluid and articular cartilage were obtained.The contents of IL-1? and NO were detected by ELISA.The degree of destruction of articular cartilage was evaluated by HE staining and Mankin's score.m RNA expressions of Wnt5 a,?-catenin,Sox9 and Collagen?were surveyed by Real-time fluorescence quantitative RT-PCR.Protein expressions of Wnt5 a,?-catenin,Sox9 and Collagen?were measured by Western blot.2 Clinical experiments: 60 cases were randomly divided into treatment group and control group with 30 cases in each group.Treatment group was given Jiawei Duhuo Jisheng Mixture,62.5 ml each time,twice a day,orally;while control group was given diacerein capsules,1 pill a day,twice a day,orally.Basic treatments were the same in both groups.Six weeks later,VAS score,WOMAC score were observed before and after treatment,so were the IL-1?and NO contents by ELISA.Results:1 Animal experiments: 1.1 ELISA results showed:(1)Compared with group A,contents of IL-1? and NO in group B were reduced obviously(P<0.05);compared with group B,contents of IL-1? and NO in group C and D were reduced obviously(P<0.05);compared with group D,contents of IL-1? and NO in group C showed no statistic difference.1.2 HE staining and Mankin score showed:(1)Compared with group A,Mankin's score in group B were reduced obviously(P<0.05);compared with group B,Mankin score in group C and D were reduced obviously(P<0.05);compared with group D,Mankin score in group C showed no statistic difference(P>0.05).1.3 RT-PCR results showed: Compared with group A,m RNA expressions of Wnt5 a and ?-catenin in group B were increased obviously(P < 0.05)while m RNA expressions of Sox9 and collagen?in group B were reduced obviously(P<0.05).Compared with group B,m RNA expressions of Wnt5 a and ?-catenin in group C and D were reduced obviously(P<0.05)while m RNA expressions of Sox9 and collagen?in group C and D were increased obviously(P<0.05).Compared with group D,m RNA expressions of Wnt5 a,?-catenin,Sox9 and collagen?in group C showed no statistic difference(P>0.05).1.4 Western blot results showed: Compared with group A,protein expressions of Wnt5 a and ?-catenin in group B were increased obviously(P<0.05)while protein expressions of Sox9 and collagen?in group B were reduced obviously(P<0.05).Compared with group B,protein expressions of Wnt5 a and ?-catenin in group C and D were reduced obviously(P<0.05)while protein expressions of Sox9 and collagen?in group C and D were increased obviously(P<0.05).Compared with group D,protein expressions of Wnt5 a,?-catenin,Sox9 and collagen?in group C showed no statistic difference(P>0.05).2 Clinical experiments: Compared with pre-treatment,VAS score,WOMAC score,contents of IL-1?,NO,Wnt5 a and ?-catenin by ELISA in both control and treatment groups were reduced obviously after treatment(P<0.05)while the content of Sox9?Collagen? were increased(P<0.05);VAS score,WOMAC score,contents of IL-1?,NO,Wnt5 a,?-catenin,Sox9 and Collagen?by ELISA showed no statistic difference in control and treatment groups after treatment(P>0.05).Conclusions: 1 Animal experiments: The KOA model with TCM pathogenesis of "deficiency,blood Stasis and toxin" was established.Inflammatory and overactive Wnt signaling pathways may be an important factor leading to cartilage destruction.Jiawei Duhuo Jisheng Mixture may promote cartilage repair via Wnt signal pathway and Sox9 may be one of its effect target.2 Clinical experiments: Jiawei Duhuo Jisheng Mixture can reduce inflammatory factors of the join fluid and may inhibit the Wnt signal pathway through regulating the expression of Sox9 and collagen ? in the pathway,which can promote the cell proliferation of knee osteoarthritis,cartilage corpuscle.Jiawei Duhuo Jisheng Mixture is a safe drug to treat KOA.