BMSCs诱导分化为胆碱能神经元过程中干扰Hes1表达对Stmn1和Sept9基因的影响  

Effects of interfering with gene Hes1on the expression of the genes Stmn1 and Sept9 inducing BMSCs differentiation into cholinergic neurons

作　　者：李鹏飞[1] 王春芳[2] 柳勇[3] 崔素梅 邓春圣[3] 龙志晶[2] 刘彩玉[2] 

机构地区：[1]山西医科大学科研实验中心,太原030001 [2]山西医科大学实验动物中心,太原030001 [3]中国人民解放军第322医院,大同037006 [4]山西康宝生物制品股份有限公司,长治046000

出　　处：《神经解剖学杂志》2013年第1期45-50,共6页Chinese Journal of Neuroanatomy

基　　金：山西省自然科学基金(2009011057-1);山西省实验动物专项资金项目(2012k01);中国人民解放军第322医院自然科学基金(2012001)

摘　　要：目的:通过检测体外培养的骨髓基质细胞(bone marrow stromal cells,BMSCs)定向诱导分化为胆碱能神经元过程中干扰Hes1基因对Stmn1和Sept9基因的表达变化情况,探讨骨髓基质细胞定向分化为胆碱能神经元的机制。方法:采用全骨髓培养法体外分离培养获得BMSCs,将第三代培养的BMSCs进行诱导分化,实时定量PCR技术分析干扰Hes1基因后,在骨髓基质细胞诱导分化为胆碱能神经元过程中Stmn1和Sept9基因的表达变化情况。结果:BMSCs诱导分化为胆碱能神经元后,经免疫荧光检测有乙酰胆碱转移酶阳性细胞表达;Hes1基因表达在诱导组和干扰后诱导组中均较对照组高(P<0.05);Stmn1基因表达水平在干扰后诱导组、干扰组、诱导组均较对照组高(P<0.05);Sept9基因在干扰后诱导组和干扰组中表达水平均较对照组高(P<0.05),诱导组和对照组间无统计学意义。结论:在BMSCs向胆碱能神经元分化过程中,干扰Hes1基因可调控Sept9,Stmn1的表达,Hes1基因的表达变化导致与分化和增殖相关基因的表达改变。Objective： To study the expression of the genes Sept9 and Stmnl in the process of cholinergic neurons differ- entiation from bone marrow stromal cells （BMSCs） after interference with Hesl gene, and to investigate the mechanisms underlying the differentiation of neural stem cells into cholinergic neurons. Methods ： BMSCs were isolated from the whole bone marrow culture in vitro. The third generation of BMSCs was used to induce differentiation of cholinergic neurons. Af- ter interference with Hesl gene in the inducing process, the expression of Stmnl and Sept9 mRNAs was examined by real- time quantitative PCR. Results： After induction, some cells were positive for choline acetyl transferase （CHAT） as re- vealed by immunofluorescence. The expression of Hesl in differentiation group and the differentiation-interference group, was higher than that in the control（ P 〈 0. 05 ）. Stmnl level in the differentiation-interference group, interference group, and differentiation group, was higher than that in the control（ P 〈 0.05 ）. Sept9 expression level in the differentiation-in- terference group and interference group, was higher than that in the control group（ P 〈 0.05 ） ; and there was no statistical significance between differentiation group and control. Conclusion： In the process of cholinergic neurons differentiation from BMSCs, interference with the gene Hesl can affect the expression of Stmnl and Sept9 genes, and the changes of Hesl gene＇s expression lead to the expression changes of differentiation- and proliferation-related genes.

关 键 词：骨髓基质细胞 胆碱能神经元 RNA干扰 Hes1基因 Stmn1基因 Sept9基因 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]