DSCAM在大鼠骨髓间质干细胞分化为神经细胞中的表达变化

发布时间：2019-03-15 09:30

【摘要】：【背景和目的】骨髓间质干细胞(marrow mesenchymal stem cells,MSCs)因具有很强的分裂、增殖及自我更新能力,传统上认为MSCs可分化为肌细胞、骨细胞、血细胞等组织细胞。近年来研究发现MSCs在一定的条件下可横向分化为神经细胞,如在有丝分裂因子、维甲酸类化合物、神经营养因子、中药等诱导下。唐氏综合症细胞粘附分子(Down syndrome cell adhesion molecule,DSCAM)的基因位于21q22,其过量表达,如21号染色体的三体化,即唐氏综合症(Downsyndrome),造成神经细胞迁移、增殖、分化的异常,最终导致先天性智能发育障碍。若DSCAM表达降低则出现神经连接异常机会增加。而且,DSCAM是神经细胞连接中必须的细胞粘附分子,在轴突和树突的生长、突触的形成、对神经网络的形成和维持有重要作用。研究唐氏综合症细胞粘附分子(DSCAM)在大鼠骨髓间质干细胞(MSCs)分化为神经细胞中的作用。 【方法】在建立黄芩苷诱导大鼠MSCs分化为神经细胞的基础上,采用免疫细胞化学法、Western Blot法等检测DSCAM的表达变化;同时采用RNA干扰技术,观察DSCAM-siRNA转染MSCs后诱导分化情况。 【结果】诱导前大鼠MSCs不表达DSCAM:预诱导24h:MSCs开始少量表达DSCAM(1.71%±0.67%);黄芩苷诱导6h,部分表达DSCAM(15.79%±4.24%);诱导后3d,DSCAM表达最高(53.16%±5.94%);诱导6d,DSCAM表达明显下降(28.99%±6.72%)。DSCAM-siRNA转染MSCs,DSCAM表达显著下降。而且,诱导前MSCs不表达神经细胞特异性标记蛋白β-Ⅲ-tubulin;诱导6h,β-Ⅲ-tubulin表达为(1.40%±0.79%)。诱导3d达到(41.59%±3.17%);诱导6d,β-Ⅲ-tubulin表达为(59.11%±4.76%)。但是,DSCAM-siRNA转染MSCs,诱导3d、6d,β-Ⅲ-tubulin蛋白的表达显著下降(28.57%±2.91%、43.90%±12.31%)。 【结论】DSCAM可能在骨髓间质干细胞(MSCs)分化为神经细胞中可能起到重要的作用。
[Abstract]:[background and objective] Bone marrow mesenchymal stem cells (marrow mesenchymal stem cells,MSCs), due to their strong ability of division, proliferation and self-renewal, have traditionally been thought to differentiate into muscle cells, bone cells, blood cells and other tissue cells, such as muscle cells, bone cells, blood cells and so on. In recent years, it has been found that MSCs can differentiate into nerve cells transversely under certain conditions, such as mitosis factor, retinoic acid compound, neurotrophic factor, traditional Chinese medicine and so on. The Down's syndrome cell adhesion molecule (Downsyndrome cell adhesion molecule,DSCAM) gene is located in 21q22, and its overexpression, such as trimerization of chromosome 21, that is, Down's syndrome (Downsyndrome), results in abnormal migration, proliferation and differentiation of nerve cells. Eventually leads to congenital mental retardation. If the expression of DSCAM decreased, the chance of abnormal nerve connection increased. Moreover, DSCAM is a necessary cell adhesion molecule in nerve cell junctions. The growth of axons and dendrites, the formation of synapses and the formation of synapses play an important role in the formation and maintenance of neural networks. To investigate the role of Down's syndrome cell adhesion molecule (DSCAM) in the differentiation of rat bone marrow mesenchymal stem cells (MSCs) into nerve cells. [methods] on the basis of establishing baicalin-induced differentiation of rat MSCs into nerve cells, the expression of DSCAM was detected by immunocytochemical method (, Western Blot) and RNA interference technique was used to observe the differentiation induced by DSCAM-siRNA transfection into MSCs. [results] A few expression of DSCAM (1.71% 卤0.67%) and partial expression of DSCAM (15.79% 卤4.24%) were observed at 6 h after induction of baicalin in MSCs (1.71% 卤0.67%) and DSCAM (15.79% 卤4.24%). On the 3rd day after induction, the expression of DSCAM was the highest (53.16% 卤5.94%), the expression of DSCAM was decreased significantly (28.99% 卤6.72%) on the 6th day after induction, and the expression of MSCs,DSCAM was significantly decreased by DSCAM-siRNA transfection. Moreover, MSCs did not express neuron specific marker protein 尾-鈪，

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