丹参素定向诱导骨髓间质干细胞分化为神经元样细胞的研究

目的 研究丹参素体外定向诱导骨髓间质干细胞(mesenchymal stem cells，MSC)分化为神经元样细胞。方法 利用Ficoll-Paque液将骨髓细胞进行密度梯度离心和贴壁筛选分离出MSC，体外扩增，流式细胞仪检测MSC表面抗原表达。采用含丹参素的无血清L-DMEM诱导MSC分化为神经元样细胞，并且探讨丹参素不同浓度及不同作用时间对MSC诱导分化为神经元样细胞的影响。以免疫组化方法检测神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF-M)、巢蛋白(nestin)、胶质纤维酸性蛋白(GFAP)的表达、结果 MSC在体外传代扩增后，流式细胞仪检测结果显示CD29、CD44、CD166表达阳性，CDl4、CD34、CD45、HLA-DR为阴性。丹参素诱导后，MSC胞体收缩，突起伸出，呈典型的核周体形态，类似神经元；免疫组化显示诱导出的神经元样细胞NSE、NF-M、nestin表达阳性，GFAP阴性。结论 丹参素可以在体外诱导MSC分化为神经元样细胞，其诱导转化率与丹参素浓度和作用时间有关。

Study on Effect of Danshensu in Directional Differentiation of Mesenchymal Stem Cells into Neuron-like Cells

OBJECTIVE: To study the effect of Danshensu (DSS) on directional differentiation of mesenchymal stem cells (MSC) into neuron-like cells. METHODS: MSC were separated from bone marrow with density gradient centrifugation, wall sticking screening and amplified in vitro. Flow cytometry was used to monitor the expression of surface antigens. DSS contained in non-serum L-DMEM was used to induce differentiation of MSC to neuronlike cells, and the effect of DSS when different concentration and acting time used was explored. And levels of neuron-specific enolase (NSE), neurofilament protein (NF-M), nestin, and expression of glial fibrillary acidic protein (GFAP) were measured by immunohistochemical method. RESULTS: After being propagated and amplified in vitro, MSC were positively expressed for CD29, CD44, CD166, and negatively expressed for CD14, CD34, CD45, HLA-DR. After induction of DSS, MSC exhibited the typical form of perikaryon with pyknotic cell body and prominence projected like that of neuron. These cells were positively expressed in NSE, NF-M and nestin, and negatively expressed in GFAP. CONCLUSION: DSS could induce differentiation of MSC to neuron-like cells in vitro, the action is concentration- and time-dependent.