【摘要】 目的　通过尼古丁对小鼠胚胎干细胞 (EmbryonicStem ,ES)特异基因Oct 4转录的影响来探讨尼古丁是否影响早期胚胎发育。方法　ES细胞选用饲养层培养法 ,尼古丁 (1～ 10 0 0nmol L)和 或 10 μmol L筒箭毒 (尼古丁乙酰胆碱受体阻断剂 )作用 2 4h后 ,纯化ES细胞 ,提取总RNA。首先扩增Fgf 5以观察纯化后ES细胞的纯度 ;同时用半定量RT PCR法观察尼古丁对Oct 4和 β actin表达的影响 ,及筒箭毒对尼古丁作用的影响。结果　尼古丁对β actin转录没有明显的影响 ,而可明显的增强Oct 4的转录 ;筒箭毒单独作用ES细胞可提高Oct 4的转录 ,而在尼古丁同时存在的情况下 ,筒箭毒对Oct 4的作用将随着尼古丁浓度的增加而减弱 ,当尼古丁的浓度为 10 0～ 10 0 0nmol L时 ,筒箭毒可明显抑制尼古丁诱导的Oct 4的表达。结论　研究表明 ,Oct 4的转录水平和胚胎干细胞的命运密切相关 ,维持胚胎干细胞的未分化状态需要Oct 4保持相对恒定的转录水平。因而 ,尼古丁对Oct 4转录的影响可能会影响胚胎干细胞的发育和分化 ,从而影响胚胎的早期发育更多还原

【Abstract】 To clarify whether nicotine affected the early embryo development, the paper investigated the influence of nicotine on embryonic stem (ES) cells specific gene Oct-4. Methods ES cells were treated with nicotine (1～1000 nM) and/or 10 μM tubocurarine (a usual nicotinic acetylcholine receptors [nAChRs] blocker) for 24 h. Subsequently, they were purified to discard feeder cells and total RNA was isolated. Fgf-5 was amplified to detect the purification of ES cells, and the relative expression of Oct-4 and β-actin to GAPDH was analyzed with RT-PCR. Results RT-PCR analysis illustrated that nicotine (10～1000 nM) significantly enhanced Oct-4 transcription, while had no effect on β-actin transcription; meanwhile, compared with nicotine (100 nM and 1000 nM) treatment alone, tubocurarine inhibited Oct-4 transcription evidently. Conclusion It is reasonable to assume that nicotine could influence the development and differentiation of ES cells, and impinge on the early embryo development. $$$$更多还原