Study On The Effect And Mechanism Of Shennao Fuyuan Decoction In Inducing The Proliferation, Mobilization And Differentiation Of BMSCs

Objective:To observe the effects of serum containing ShenNaoFuYuan Decoction(SNFYD)on the proliferation,migration and differentiation of rat bone marrow mesenchymal stem cells(BMSCs)and its effect on the expression of SDF-1/CXCR4 cytokine,and provide the experimental basis for elucidating the possible mechanism of ischemic stroke and elucidating its clinical efficacy.Methods:1,the acquisition and identification of BMSCs: strictly aseptic operation to obtain and expose the bone marrow cavity of standard SD rats,DMEM-HG repeatedly rinse,blow after centrifugation,collect cells,put in the double anti DMEM-HG medium of 10%FBS and culture,and use flow cytometry to identify rat bone marrow mesenchymal stem cells.2,SNFYD containing serum culture medium for culture and generation of BMSCs: use qualified SD rats to prepare the medicine serum of SNFYD,prepare culture liquid with different concentration of serum containing medicine,and set up ordinary culture liquid and blank sera as normal control group and blank control group,and compare and analyze the culture of BMSCs in culture medium.3,Detection of related data: under electron microscope,BMSCs morphology,CCK8 method were observed to detect the proliferation of BMSCs,Transwell method was used to detect the migration ability,NSE immunohistochemical method was used to determine the differentiation ability,and the Western blot method was used to detect the expression of CXCR4 and SDF-1 molecular protein.Results:Compared with the control group,the number of BMSCs in the serum culture solution group increased significantly,the morphology was closer to the spindle and fibroblast like structure,the amplification ability was enhanced,the expression rate of NSE increased,and the expression of CXCR4 and SDF-1 increased.These results suggest that the drug serum of SNFYD can enhance the proliferation of bone marrow mesenchymal stem cells and induce its differentiation into neuron cells.This effect may be related to the up-regulation of CXCR4 and SDF-1.Conclusions:1.The serum containing SNFYD can obviously affect the physiological process of BMSCs,which is mainly reflected in accelerating its formation and proliferation,promoting its differentiation to nerve like cells and enhancing the migration ability.2.The expression of SDF-1/CXCR4 in the sera cultured with SNFYD increased.At present,it is clear that SDF-1/CXCR4 signaling pathway has the role of promoting vascular regeneration and brain protection in the region of brain necrosis.The effect of kidney brain compound decoction on BMSCs may be related to the activation of the cell pathway.3.At present,the drug serum cell experiment of SNFYD supports the theory of "renal medullary through the brain",and the follow-up should be based on this theory to design animal experiments to verify the effect and effect of kidney brain compound soup in the mobilization of endogenous BMSCs in vivo,and further improve the experimental basis of the theory of "renal medullary through brain".