三七总皂苷在骨髓间质干细胞分化为神经元样细胞中的调解作用研究

目的:探讨三七总皂苷在骨髓间质干细胞(MSCs)分化为神经元样细胞中的调解作用。方法:①实验材料:成年健康Wister大鼠(雌雄不限),SRF级,体质量(150±30)g;②实验方法:无菌的条件下从大鼠的股骨中分离并继代培养MSCs细胞,当传至第5代时,先用完全培养液含10μg/L碱性成纤维细胞生长因子(bFGF)]进行预诱导,再更换无血清培养液(含三七总皂苷0.25g/L)进行诱导,采用免疫组织化学SABC法检测神经丝蛋白(NF-M)、神经元特异性烯醇化酶(NSE)、巢蛋白(nestin)、微管联合蛋白-2(MAP-2)、生长相关蛋白43(GAP-43)以及胶质纤维酸性蛋白(GFAP)的表达。结果:①采用三七总皂苷诱导后,大多数MSCs呈神经元样细胞分化。经免疫组织化学鉴定,上述细胞表现为NSE、MAP-2和NF染色阳性,呈棕黄色,而GFAP染色阴性。②采用三七总皂苷进行诱导时,MSCs的胞体逐渐增大,并伸出细长的突起,在突起的末端出现分支,有些相邻细胞的突起还成网状连接。③三七总皂苷诱导MSCs转变为类似于神经元样的形态,经免疫组织化学鉴定,大多数细胞表现为NF-M、NSE、nestin、MAP-2和GAP-43染色阳性,并在胞质和突起中有棕黄色的物质,而GFAP染色均为阴性。SABC法显示NSE和MAP-2阳性细胞数分别为(72.64±2.04)%和(73.5±2.17)%。结论:三七总皂苷在体外能使MSCs向神经元样细胞转化,且具有神经元样细胞的特性,这为MSCs治疗相关疾病提供了理论基础。

Study on the Mediating Role of PNS in Bone Marrow Mesenchymal Stem Cells Differentiating into Neuron - like Cells

Objective:To probe into the mediating role of PNS in bone marrow mesenchymal stem cells differentiating into neuron-like cells.Methods:①Experimental material: Healthy adult Wister rats(male or female),SRF level,weighted(150±30)g.②Experimental Methods: In sterile conditions,the isolated rat MSCs cells femur were subcultured,on the 5th generation,containing 10μg/L basic fibroblast growth factor(bFGF) in complete culture was for pre-induction medium,then replace by the PNS 0.25g/L of serum free medium to induce.Immunohistochemical SABC assay was used to test neurofilament(NF-M),neuron-specific enolase(NSE),nestin(nestin),united microtubule protein-2(MAP-2),growth associated protein 43(GAP-43),and glial fibrillary acidic protein(GFAP) expression.Results:①MSCs used for induction of PNS,the majority of MSCs were neuron-like cells.Identified by immunohistochemistry,these differentiated cells showed NSE,MAP-2 and NF staining,cells were brown,and GFAP was negative.②Induced by PNS,the mesenchymal stem cells increase cell bodies and extended long thin processes,a branch in the protruding end of the emergence of the sudden adjacent cells to form a network connection can also be played.③PNS induced mesenchymal stem cells into neuron-like morphology,identified by immunohistochemistry,the majority of cells showed NF-M,NSE,nestin,MAP-2 and GAP-43 staining,and cell quality and processes in the brown substance,and GFAP staining was negative.SABC method showed that NSE and MAP-2 positive cells were(72.64±2.04)% and(73.57±2.17)%.Conclusions:Inducer of bone marrow MSCs in vitro effect of PNS neuron-like cells have been transformed,and has the characteristics of neuron-like cells,which will promote the application of bone marrow MSCs as a treatment of diseases related to cell engineering and provide a theoretical basis.