The Regulatory Mechanism Research Of Chinese Medicine Articular Cavity Perfusion To MSCs In Steroid-induced Avascular Necrosis Of The Femoral Head

Objective:By building the rabbit model of steroid-induced avascular necrosis of the femoral head,we explore the action mechanism on two aspects of the results.On the one side,observe and investigate the influence of the traditional Chinese medicine HuoGU injection working on the mesenchymal stem cells' proliferation and differentiation.On the other side,research the repair function of the HuoGU injection injecting cooperate with MSCs transplanting to the femoral head of early SANFH rabbits.Methods:(1)Select the adult male rabbits,build the steroid-induced avascular necrosis model.Extract the mesenchymal stem cells.Divide the Experimental animals into three groups:experimental group,control group and blank group.Describe proliferation curve determined by MTT,calculate the activity of ALP and calcified nodules.Test three sets of transfection MSCs migration ability.Use RT-PCR to detect the MMP-2 mRNA.Observe and investigate the influence of the traditional Chinese medicine HuoGU injection working on the mesenchymal stem cells' proliferation,differentiation and migration.(2)Randomly divide the rabbits into four groups:blank group,model group,control group and experimental group.Control group is simply transplanted MSCs to necrotic area.Experimental group uses HuoGU injection cooperate with MSCs transplanting to treat.Use immunohis-tochemical SABC method to detect Protein expression of MMP-2,t VEGF,TGF-?1;RT-PCR detects the MMP-2 mRNA,VEGF mRNA,TGF-?1 mRNA expression.Von Kossa staining observes the calcium nodules appear time and quantity;HE staining observe the repair of the femoral head under the microscope,research the influence of the traditional Chinese medicine HuoGU injection to transplanted MSCs migration,proliferation and osteogenesis differentiation.Results:the method of combined use of horse serum and methylpred-nisolone can successfully cause the SANFH rabbit model.HuoGu injection can effectively promote the MSCs proliferation,promote MSCs different-tiation to osteogenesis,compared with the rest of the groups,the difference is statistically significant(P<0.05).RT-PRC detection of MMP-2 mRNA experimental results show that HuoGu injection can promote the MSCs migration ability.In vivo tests results show that HuoGu injection can effectively improve the MMP-2,VEGF,TGF-?1 within the femoral head in both protein and mRNA expression It promotes the MSCs osteogenesis differentiation,migration in the body and promotes restoration of the necrotic femoral head,efficacy significantly(P<0.05).Conclusion:Horse serum cooperates with hormone intervention can simulate early pathological changes of SANFH of people.HuoGu injection promotes the MSCs differentiation,proliferation,migration and increase the femoral head local blood circulation,promote restoration of the femoral head effect,effectively prevents the progress of the disease.By discussing the traditional Chinese medicine of SANFH MSCs mechanism research,making deeper understanding of SANFH early pathological changes.Select one of the key links to intervene,put forward a better treatment to SANFH.alleviate the pain of patients,improve the quality of life.