川芎嗪体外诱导小鼠骨髓间充质干细胞分化为神经元样细胞的研究  

Differentiation of mouse bone marrow mesenchymal stem cells into neuron-like cells induced by tetramethylpyrazine in vitro

作　　者：刘云云[1] 赵兴绪[1] 赵红斌[2] 葛宝丰[2] 刘兴炎[2] 陈克明[2] 

机构地区：[1]甘肃农业大学生命科学学院,兰州730070 [2]兰州军区兰州总医院骨科研究所,兰州730050

出　　处：《生物学杂志》2010年第6期21-24,36,共5页Journal of Biology

基　　金：甘肃省自然科学基金(3ZS051-A25-078)

摘　　要：为了探讨川芎嗪体外诱导小鼠骨髓间质干细胞(BMSCs)分化为神经元样细胞的作用,以小鼠骨髓间充质干细胞为研究对象,实验分为空白对照组、β-巯基乙醇(BME)阳性对照组和川芎嗪诱导组。采用荧光免疫化学和Western blot方法,分别检测神经干细胞巢蛋白(nestin)和经元特异性烯醇化酶(NSE)的表达;RT-PCR检测诱导不同时间对神经细胞相关基因Nestin、NSE、β-微管蛋白III(β-Tubulin III)和核受体相关因子-1(Nurr1)mRNA表达的影响。结果显示川芎嗪诱导间充质干细胞24 h后,细胞形态发生显著改变,细胞突起形成且数目不等,形成神经元样细胞。细胞死亡率低于β-巯基乙醇诱导组。免疫荧光化学法和western blot结果显示:川芎嗪诱导后的细胞nes-tin和NSE蛋白表达呈阳性,且表达丰度显著高于β-巯基乙醇诱导组。川芎嗪作用不同时间的BMSCs表达神经细胞相关基因Nestin、β-Tubulin III、NSE和Nurrl。结果表明川芎嗪能定向诱导小鼠骨髓间充质干细胞分化为神经元样细胞,是较理想的诱导剂。The effect of tetramethylpyrazine on inducing differentiation of mouse bone marrow-derived mesenchymal stem cells（BMSCs） into neuron-like cells was studied in vitro.The experiments were divided into 3 groups such as the control group,β-mercaptoethanol induction group and the tetramethylpyrazine induction group.Flourescence immunohistochemistry and Western blot were used to detect several neural specific proteins such as nestin and NSE.The related genes of nerve cells,Nestin,β-Tubulin III,NSE and Nurr1,were deteced by RT-PCR method in different times.The induced MSCs by tetramethylpyrazine gradually exhibited neuronal morphological characteristics.The mortality rate of MSCs caused by tetramethylpyrazine was much lower than by β-mercaptoethanol.The expression of several neural specific proteins like nestin and neuron-specific enolase（NSE） was detected by immunocytochemistry and western blot methods.The level of these proteins in BMSCs induced by tetramethylpyrazine were much higher than by β-mercaptoethanol.RT-PCR analysis revealed that an expression of several neural genes including Nestin,β-Tubulin III,NSE nd Nurrl was found in the induced BMSCs.These results suggested that the mouse bone marrow-derived mesenchymal stem cells could be differentiated into neuron-like cells in vitro with the induction of tetramethylpyrazine.

关 键 词：川芎嗪 骨髓间充质干细胞 神经元样细胞 细胞分化 

分 类 号：Q813.5[生物学—生物工程] R282.71[医药卫生—中药学]