电磁场在转录水平对胚胎干细胞源性神经前体细胞凋亡相关基因影.doc

电磁场在转录水平对胚胎干细胞源性神经前体细胞凋亡相关基因影 　 作者：秦雅鑫，宋琦，殷樱，许川山 【摘要】 目的： 利用小鼠胚胎干（ES）细胞分化成神经前体细胞模型，研究极低频率电磁场（ELFEMF）对细胞周期调节基因、凋亡相关基因以及细胞增生和凋亡的影响. 方法： 分离、培养和诱导ES成神经前体细胞，于诱导后第4日起，实验组予ELFEMF干预，对照组采用假暴露，于4+4 d, 4+7 d，4+11 d，4+17 d，4+23 d使用BrdU和Nestin免疫荧光染色检测细胞生长分化，RTPCR检测bcl2，bax，GADD45基因转录产物水平. 结果： 对照组和实验组的BrdU和Nestin免疫荧光染色未能发现细胞生长和分化有明显改变，对照组和实验组的RTPCR检测发现，在4+11 d时bcl2和bax mRNA 表达升高(Plt;0.01; Plt;0.05). GADD45转录水平在终末期（4+23 d）下降(Plt;0.01). 结论： EMF能够暂时性地影响ES源性神经前体细胞凋亡和细胞周期调控相关基因的转录水平，但这些改变不足以使细胞发生可检测到的细胞生理上改变. 【关键词】 小鼠; 胚胎; 干细胞；低频率电磁场；基因, bcl2；bax；GADD45 　　【Abstract】 AIM: To investigate the effect of electromagnetic fields on the cell cycle regulatory gene and apoptosisrelated gene and on cells proliferation and apoptosis with the model of mouse embryonic stem cells (ESCs) differentiated into neural progenitor cells. METHODS: ESCs were cultured and differentiated into neural progenitor cells; after 4 d of differentiation, the experimental group was exposed to extremely low frequency electromagnetic fields, while the control group was shamexposure. Immunofluorescence of BrdU and Nestin was used to detect the development and differentiation, and RTPCR was applied to detect the changes of transcript levels of bcl2, bax, GADD45 gene at the day (4+4), (4+7), (4+11), (4+17), and (4+23). RESULTS: No significant difference was found in the experimental and control groups in cell growth and differentiation. ELFEMF induced upregulation of bcl2 and bax at d (4+11) (Plt;0.01; Plt;0.05) and downregulation of GADD45 at d (4+23) (Plt;0.01). CONCLUSION: EMF exposure of ESCsderived neural progenitor cells transiently affects the transcript level of genes related to apoptosis and cell cycle control, and these responses are not associated with detectable changes of cell physiology. 　　【Keywords】 mic embryo; stem cells; low frequency electromagnetic fields; genes bcl2; bax; GADD45 　　0引言 　　多能胚胎干（ES）细胞可以分化成三个胚层的特异性细胞. 在分化的过程中，ES细胞再现了早期胚胎的发育过程，因此这提供了一个极好的体外分析遗传和环境因素对细胞分化影响的模型［1］. 极低频率电磁场（EL