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Expression changes of Notch-related genes during the differentiation of human mesenchymal stem cells into neurons
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- Author:
- No author available
- Journal Title:
- ACTA PHYSIOLOGICA SINICA
- Issue:
- 3
- DOI:
- 10.3321/j.issn:0371-0874.2007.03.004
- Key Word:
- 骨髓间充质干细胞;Notch1;JAG1;早老素1;HES1;分化;神经细胞
Abstract： 本研究探讨Notch信号通路在人骨髓间充质干细胞(human mesenchymal stem cells,hMSCs)体外增殖及向神经细胞分化过程中的作用.采集健康自愿者骨髓,体外培养获得hMSCs,取第3代hMSCs,在诱导剂(β-ME,DMSO,BHA)作用下向神经细胞分化.诱导后用免疫细胞化学鉴定神经元特异性烯醇化酶(neuron-specific enolase,NSE)和尼氏体的表达以确定诱导效果;用流式细胞术检测细胞生长周期时相的变化.在诱导前后,用免疫荧光和RT-PCR 方法检测Notch通路中Notch1受体蛋白、配体Jagged1(JAG1)、调节蛋白活化相关物早老素1(presenilin 1,PS1)、靶基因hairy and enhancer of split1(HES1)信号分子表达的变化.结果显示:诱导前,处于G0/G1期的hMSCs占58.5%,S+G2/M 期的细胞占41.5%;诱导后,G0/G1期细胞比例升高,而S+G2/M期细胞比例下降,NSE阳性细胞率达(77±0.35)%,细胞质中可见深蓝色的块状或颗粒状尼氏体.免疫荧光显示,诱导前后hMSCs内Notch1和JAG1均呈阳性表达,但RT-PCR 检测发现诱导后Notch1、JAG1、PS1和HES1 mRNA表达量较诱导前明显降低(均P＜0.05).结果表明,诱导hMSCs向神经细胞分化能抑制Notch信号分子表达,低水平的Notch信号激活可能有利于神经细胞的分化.
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