發(fā)布時(shí)間：2019-03-15 09:30

【摘要】：【背景和目的】骨髓間質(zhì)干細(xì)胞(marrow mesenchymal stem cells,MSCs)因具有很強(qiáng)的分裂、增殖及自我更新能力,傳統(tǒng)上認(rèn)為MSCs可分化為肌細(xì)胞、骨細(xì)胞、血細(xì)胞等組織細(xì)胞。近年來研究發(fā)現(xiàn)MSCs在一定的條件下可橫向分化為神經(jīng)細(xì)胞,如在有絲分裂因子、維甲酸類化合物、神經(jīng)營養(yǎng)因子、中藥等誘導(dǎo)下。唐氏綜合癥細(xì)胞粘附分子(Down syndrome cell adhesion molecule,DSCAM)的基因位于21q22,其過量表達(dá),如21號(hào)染色體的三體化,即唐氏綜合癥(Downsyndrome),造成神經(jīng)細(xì)胞遷移、增殖、分化的異常,最終導(dǎo)致先天性智能發(fā)育障礙。若DSCAM表達(dá)降低則出現(xiàn)神經(jīng)連接異常機(jī)會(huì)增加。而且,DSCAM是神經(jīng)細(xì)胞連接中必須的細(xì)胞粘附分子,在軸突和樹突的生長、突觸的形成、對神經(jīng)網(wǎng)絡(luò)的形成和維持有重要作用。研究唐氏綜合癥細(xì)胞粘附分子(DSCAM)在大鼠骨髓間質(zhì)干細(xì)胞(MSCs)分化為神經(jīng)細(xì)胞中的作用。 【方法】在建立黃芩苷誘導(dǎo)大鼠MSCs分化為神經(jīng)細(xì)胞的基礎(chǔ)上,采用免疫細(xì)胞化學(xué)法、Western Blot法等檢測DSCAM的表達(dá)變化;同時(shí)采用RNA干擾技術(shù),觀察DSCAM-siRNA轉(zhuǎn)染MSCs后誘導(dǎo)分化情況。 【結(jié)果】誘導(dǎo)前大鼠MSCs不表達(dá)DSCAM:預(yù)誘導(dǎo)24h:MSCs開始少量表達(dá)DSCAM(1.71%±0.67%);黃芩苷誘導(dǎo)6h,部分表達(dá)DSCAM(15.79%±4.24%);誘導(dǎo)后3d,DSCAM表達(dá)最高(53.16%±5.94%);誘導(dǎo)6d,DSCAM表達(dá)明顯下降(28.99%±6.72%)。DSCAM-siRNA轉(zhuǎn)染MSCs,DSCAM表達(dá)顯著下降。而且,誘導(dǎo)前MSCs不表達(dá)神經(jīng)細(xì)胞特異性標(biāo)記蛋白β-Ⅲ-tubulin;誘導(dǎo)6h,β-Ⅲ-tubulin表達(dá)為(1.40%±0.79%)。誘導(dǎo)3d達(dá)到(41.59%±3.17%);誘導(dǎo)6d,β-Ⅲ-tubulin表達(dá)為(59.11%±4.76%)。但是,DSCAM-siRNA轉(zhuǎn)染MSCs,誘導(dǎo)3d、6d,β-Ⅲ-tubulin蛋白的表達(dá)顯著下降(28.57%±2.91%、43.90%±12.31%)。 【結(jié)論】DSCAM可能在骨髓間質(zhì)干細(xì)胞(MSCs)分化為神經(jīng)細(xì)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% 鹵0.67%) and partial expression of DSCAM (15.79% 鹵4.24%) were observed at 6 h after induction of baicalin in MSCs (1.71% 鹵0.67%) and DSCAM (15.79% 鹵4.24%). On the 3rd day after induction, the expression of DSCAM was the highest (53.16% 鹵5.94%), the expression of DSCAM was decreased significantly (28.99% 鹵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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