[刺五加体外诱导骨髓基质细胞向神经元样细胞分化的作用]
[Effects of Ciwujia in inducing marrow stromal cell differentiation into neuron-like cells in vitro].
作者信息
Yang Qin, Yang Jun, Xie Peng, Zeng Zhi-Lei, Li Ao, Yang Jun, Zhang Xiao-Dong, Li Yong-Mei, Lü Fa-Jin
机构信息
Department of Neurology, First Affiliated Hospital, Key Laboratory of Neurology of Chongqing, First Affiliated Hospital, Chongqing Medical University, Chongqing, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2009 Mar;29(3):487-90.
OBJECTIVE
To study the effect of the traditional Chinese herbal drug Ciwujia in inducing the differentiation of marrow stromal cells (MSCs) into neuron-like cells.
METHODS
Rat MSCs isolated from the whole bone marrow were amplified by adherent culture in vitro and induced to differentiate into neuron-like cells using serum-free DMEM/F12 containing Ciwujia. The protein and mRNA expressions of nestin, beta-Tubulin III and glial fibrillary acidic protein (GFAP) in the differentiated cells were detected by indirect immunofluorescence method, Western blotting and RT-PCR.
RESULTS
The third-passage MSCs showed positive expression rates for CD44 and CD54 beyond 90% with decreased CD14 expression rate to 2.37%. Induction by Ciwujia of the MSCs resulted in cell body shrinkage and protrusion of the cell processes resembling those of neurons. The differentiated cells were positive for nestin and beta-Tubulin III expression and negative for GFAP as shown by immunofluorescence assay, Western blotting and RT-PCR.
CONCLUSION
Ciwujia can induce the differentiation of rat MSCs into neuron-like cells in vitro.
目的
研究中药刺五加诱导骨髓基质细胞（MSCs）向神经元样细胞分化的作用。
方法
从全骨髓中分离大鼠MSCs，通过体外贴壁培养进行扩增，并用含刺五加的无血清DMEM/F12诱导其分化为神经元样细胞。采用间接免疫荧光法、蛋白质免疫印迹法和逆转录-聚合酶链反应（RT-PCR）检测分化细胞中巢蛋白、β-微管蛋白III和胶质纤维酸性蛋白（GFAP）的蛋白质和mRNA表达。
结果
第三代MSCs的CD44和CD54阳性表达率超过90%，而CD14表达率降至2.37%。刺五加诱导MSCs后，细胞体收缩，出现类似神经元的细胞突起。免疫荧光分析、蛋白质免疫印迹法和RT-PCR结果显示，分化细胞中巢蛋白和β-微管蛋白III表达呈阳性，而GFAP表达呈阴性。
结论
刺五加可在体外诱导大鼠MSCs分化为神经元样细胞。
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