丹参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:探讨丹参注射液在体外诱导大鼠骨髓间充质干细胞(MSCs)向神经元样细胞分化的作用。方法:采用差速贴壁法体外分离培养大鼠MSCs,流式细胞仪检测其表面标志,经碱性成纤维细胞生长因子(bFGF)预诱导24h,采用含丹参注射液的无血清L-DMEM培养液诱导MSCs,倒置显微镜下观察细胞形态的变化,免疫荧光细胞化学检测巢蛋白、神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF-M)、胶质纤维酸性蛋白(GFAP)的表达。结果:体外培养的大鼠MSCs表达CD29、CD44、CD105、CD166。丹参注射液诱导后MSCs胞体收缩,突起伸出,较密的部分神经元拉成网状,免疫细胞化学显示巢蛋白、NSE、NF-M表达阳性,阳性率分别为(58.68±4.50)%、(61.23±5.72)%、(63.47±6.38)%,GFAP阳性细胞率(1.47±0.23)%。结论:丹参注射液在体外可以将大鼠MSCs诱导分化为神经元样细胞。

Differentiation of Rats'' Bone Marrow Mesenchymal Stem Cells into Neuron-like Cells Induced by Salvia Miltiorrhiza Injection in Vitro

OBJECTIVE：To investigate the differentiation of rats＇bone marrow mesenchymal stem cells（MSCs） into neuron- like cells induced by Salvia miltiorrhiza injection in vitro.METHODS： MSCs which were separated from adult rats＇ bone marrow and cultured with wall sticking method in vitro were identified by flow cytometry to detect its surface marker.MSCs were pre - induced for 24 h by bFGF then induced by Salvia miltiorrhiza injection and serum - free L - DMEM culture solution.The morphological changes of MSCs were evaluated by microscope. And nestin, neuronspecific enolase （NSE）, neurofilament protein（NF-M）, glial fibrillary acidic protein（GFAP） were detected by immunoeytochemistry. RESULTS： MSCs which were positive for CD29, CD44, CD105 and CD166 were induced by Salvia miltiorrhiza injection to display neuronal morphologies, such as contracted muhipolar cell body and formed extensive networks, and express neuronal phenotype similar to the cell appearance of neurons with nestin, NSE and NF- M positive.The percentages of positive nestin,NSE and NF-M expression were（58.68 ±4.50） %, （61.23 ±5.72） % and （63.47 ± 6.38） % respectively, and the positive rate of GFAP was（1.47± 0.23）% .CONCLUSION： MSCs can be induced to differentiate into neuron- like cells in vitro by Salvia rniltiorrhiza injection.