心脉隆注射液诱导大鼠骨髓间充质干细胞分化为神经元样细胞的研究

目的 探讨心脉隆注射液体外诱导大鼠骨髓间充质干细胞(BMSCs)向神经元样细胞分化的可行性。方法 采用贴壁培养法分离纯化大鼠骨髓间充质干细胞，流式细胞仪检测细胞表面抗原的表达，免疫细胞荧光法检测诱导后巢蛋白(Nestin)、神经元特异性烯醇化酶(NSE)、微管相关蛋白2(MAP2)、胶质纤维酸性蛋白(GFAP)的表达。结果 流式细胞仪检测显示CD29、CD90阳性，CD34、CD45阴性。实验组诱导4、12h后Nestin阳性表达率分别为(81.0±1.6)%、(22.5±1.9)%，36h后Nestin阴性；诱导4h后NSE、MAP2阴性，12、36h后NSE阳性表达率分别为(73.5±2.2)%、(94.3±1.8)%，MAP2阳性表达率分别为(80.0±2.2)%、(96.4±2.8)%；诱导4、12、36h后GFAP阴性。结论 心脉隆注射液可在体外快速、高效地诱导大鼠骨髓间充质干细胞向神经元样细胞分化。

Differentiation of rat bone marrow mesenchymal stem cells into neuron-like cells induced by Xinmailong injection

Objective To explore the feasibility of Xinmailong injection inducing rat bone marrow mesenchymal stem cells(BMSCs) to differentiate into neuron-like cells.Methods The rat BMSCs were separated and purified with the adherent culture method.Following passages,the cell surface markers CD29,CD90,CD34 and CD45 were measured by flow cytometry.The expressions of Nestin,Neuron specific enolase(NSE),Microtubule associated protein-2(MAP2) and Glial fibrillary acidic protein(GFAP) were detected by immunofluorescence assay.Results Flow cytometry showed positive expression of CD29(99.06%) and CD90(99.61%) and negative expression of CD34(2.03%) and CD45(2.00%).At 4 h,12 h after induction,the positive rates of Nestin were(81.0±1.6)%,(22.5±1.9)%,while at 36 h after induction,Nestin was negative.At 4h after induction,NSE and MAP2 were negative.At 12 h,36 h after induction,the positive rates of NSE were(73.5±2.2)%,(94.3±1.8)% and those of MAP2 were(80.0±2.2)%,(96.4±2.8)%.At 4 h,12 h,36 h after induction,GFAP was nagative.Conclusion Xinmailong injection can rapidly and efficiently induce rat BMSCs to differentiate into neuron-like cells.