Experimental Study, Spleen And Kidney Side Control Rabbit Femoral Head Necrosis

1 Objective:To study the role of an effective way and target of the therapy of JianPiBuShen Tang for SINFH.To guide the clinical application,improve the clinical efficacy and to play the advantages of Chinese medicine for prevention and treatment of SINFH.2 Method:50 rabbits were randomly assigned to 5 groups(n=10):blank control,pathologic model group,JianPiBuShen treat group(high,medium,low dosage group).Model group and observation group copy the model of SINFH through gluteal injection of methylprednisolone sodium succinate(20mg/kg/d)for 3 consecutive days. At the same time,prevent the infection by intramuscular injection of penicillin.All treatment groups were high(5.52g/kg/d),medium(2.76g/kg/d),low(1.38g/kg/d) doseage administered for JianPiBuShen Tang For 12 weeks respectively.Model group and blank control group administered distilled water(5g/kg/d) for 12 weeks.TG, CHO,HDL,LDL and PT,APTT,AT-Ⅲwere detected 12 hours after oral administration at 1,4,8 and 12 weeks after the first administration.Executed all animals after the last blood sampling.Bilateral femoral heads of HE staining were observed the changes of trabecular bone,bone cells,and fat cells from microscope. Calculated the ratio of empty osteocytes lacuna and adipose tissue area.Observe collagen distribution in cartilage and trabecular bone and calculate the area of green staining of collagen by Masson staining of paraffin sections.The medicated serum was extracted from the animals of high-dose group and blank control group.2 rabbits randomly taked from the blank group were used for MSCs isolating.The 3rd generation cells which had been identificated and purificated were divided into 4 groups(JianPiBuShen Osteogenesis group,Osteogenesis group, JianPiBuShen group and blank control group).Each group cultured for corresponding medium.Respectively,after induction of 2,4,8,12,16 days,the ALP activity of each group was tested.Specimens in each group were marked with tetracycline staining 22 days later.Count the number of nodules of calcium.All statistical analysis was performed with SPSS 12.0.The data were expressed as means±standard deviation value((?)±S),and statistical significance was considered if P＜0.05.Comparison of measurement data between groups used one-way ANOVA.Inter-group comparison of the rate of osteonecrosis of the femoral head used chi-square test.3 Results:JianPiBuShen Tang can significantly improved lipid metabolism disorders and coagulopathy of the model animals caused by glucocorticoid.And pathology results showed that:JianPiBushen Tang can effectively reverse the pathological changes in rabbits SINFH.The JianPiBuShen Tang can promote the differentiation of MSCs to osteoblasts.4 Conclusion:4.1 Three consecutive high-dose LPS injection in rabbits are able to replicate the SINFH model.It is a simple and easy way to implement and has low mortality.The SINFH model copy in line with the early pathological changes of human SINFH.It is recommended. 4.2 High-dose LPS injection in rabbits could result in lipid metabolism disorder and blood into a hypercoagulation state.JianPiBuShen Fang can improve lipid metabolism and the hypercoagulable state.It can prevent the formation of fat emboli and thrombosis in local micro-circulation,and the occurrence of SINFH.4.3 The JianPiBuShen medicated serum can promote the differentiation of rMSCs to osteoblast,which may reveal cellular mechanism of prevention and treatment of SINFH.