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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 detect the changes of Stmn1 and Sept9 gene expression in bone marrow stromal cells (BMSCs) induced by differentiation into cholinergic neurons in vitro and to explore the role of bone marrow stromal cells in differentiation into gall bladder Mechanism of Alkaline Neurons. Methods: BMSCs were isolated and cultured in vitro by whole bone marrow culture. BMSCs cultured in the third generation were induced to differentiate. After real-time quantitative PCR analysis of Hes1 gene, Stmn1 and BMSCs were induced to differentiate into cholinergic neurons Sept9 gene expression changes. Results: After BMSCs differentiated into cholinergic neurons, the expression of acetylcholinesterase-positive cells was detected by immunofluorescence. The expression of Hes1 gene was higher in the induced group and the induced group than in the control group (P <0.05). Stmn1 gene (P <0.05). The expression level of Sept9 gene in the induction group and the interference group was higher than that in the control group (P <0.05), while the induction group There was no significant difference between control group and control group. Conclusion: In the process of BMSCs differentiating into cholinergic neurons, the interference of Hes1 gene can regulate the expression of Sept9 and Stmn1. The change of Hes1 gene leads to the change of the expression of genes related to differentiation and proliferation.