| ObjectiveIn researching of the transformation resistant control of the Bone marrow mesenchymal stem cell (MBCs) to lipocyte. to discuss the function of Jianpibushen fomula in curing the disease of Femur Head Necrosis and clinic feasibility. With the perspective of cytomorphological Oil-red O staining and PPAR Y formation, to identify the influence of transformation from the MBCs to lipocyte on testing rabbit caused by Jianpibushen fomula carrier blood serum, under the induce adipogenic differentiation.Method1Draw high density MBCs from testing rabbit by utilizing density gradient centrifugation with lymphocyte Percoll in the density of1.073g/ml. Then cultivate and select best P2cells of the third generation as Target cell.2Preparing the Jianpibushen fomula carrier blood serum:4Sterilized New Zealan rabbit average age of5; Jianpibushen fomula provided by the Guang Anmen Hospital decoction pharmacy department. The constitution of the Jianpibushen fomula is Fuling. Baizhu, Gancao, Niuxi. Buguzhi, Gusuibu, Yuanhu, Honghua. prescribed as1g/ml for each; Two rabbits were treated with Jianpibushen fomula earner blood serum5.52g/kg/d(which is equivalents to4.6times dosage to adult weighs60kg) by gavage once a day for7days. The rest of two rabbits were treated with distilled water with same volume by gavage. Collect blood sample accordingly and treat with Centrifugal separation to obtain serum from the upper level. Then store the serum at-20?for latter operation.3Induce adipogenic differentiation of MBCs by doing The classic adipocyte-induced system including0.1?mol/L dexamethasone,0.1mmol/L IBMX.0.1mmol/L indomethacin, and lOmg/L insulin. At the same, formula carrier Group be added with formula carrier blood serum, non interfered Group be added with blank blood serum and Control Group be added with physiological saline, adjust to a density of20%.4With the perspective of cytomorphological, to exam the obtained MBCs by induce of Fat cell differentiation, inorder to confirm the obtained cells are Bone marrow mesenchymal stem cells. 5Through the Observsion of cytomorphology, Oil-red O staining, rationing OD, and PPAR? by ELISA, to identify the differences between the formula carrier Group, the non interfered Group and the Control Group.6References were displayed with Mean and Standard deviation and All statistical analysis was performed with SPSS12.0. The data were expressed as means±standard deviation value (x±S), and statistical significance was considered if P<0.05. Comparison of measurement data between groups used one-way ANOVA. And Inter-group comparisons were analyzed with SNK-q test.Results1Density gradient centrifugation ensure high density rabbit MBCs with trusted Heterogeneity: Passage cell were grown in Uniform distribution in the form of fusiform and Long spindle. With the Induced agent, considerable quantities fat droplets were formed, it is convinced that the cultivated cell is the Bone marrow mesenchymal stem cells.2Red oil O test results:the significance between the formula carrier Group, the non interfered Group and the Control Group are statistical considerable(F=59.12, P<0.0001). The formula carrier Group<the non interfered Group, the Control Group(P<0.05), the significance between the non interfered Group and the Control Group is not statistical considerable (P>0.05). ELISA results:the PPAR? significance between the formula carrier Group, the non interfered Group and the Control Group are statistical considerable(F=33.04, P<0.0001). The formula carrier Group<the non interfered Group, the Control Group(P<0.05), the significance between the non interfered Group and the Control Group is not statistical considerable (P>0.05).Conclusion Jianpibushen fomula carrier blood serum is capable of restrain the rabbit's MBCs from Adipogenic differentiation. Together with the proved capability of Jianpibushen fomula in facilitating rabbit's MBCs Osteogenic differentiation, they demonstrate the cellular mechanism in curing the disease of the avascular necrosis of the femoral head. |
PDF Full Text Request