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目的观察组蛋白去乙酰化酶(HDAC)抑制剂曲古抑菌素A(TSA)对鸡胚翅芽发育中某些基因表达水平的作用,探讨不同浓度TSA(75μmol/L、750μmol/L、1.5mmol/L)对鸡胚翅芽发育的影响,提高我们对染色质结构重组和表观遗传对基因表达的调控作用,以及对正在开发的抗癌药物的认识。方法以鸡胚翅芽作为实验模型,用TSA处理翅芽,对鸡胚胎实施原位杂交,检测与肌肉发育相关基因的表达,并采用硫酸耐尔蓝(Nile blue sulfate)法检测细胞凋亡状况。结果75μmol/L TSA能够抑制与肌肉发生相关的基因MyoD和Myf5在鸡胚翅芽的表达,经TSA(75μmol/L)处理的胚胎没有发生明显的细胞凋亡。但是随着TSA药物浓度的提高,TSA(≥750μmol/L)不仅强烈抑制相关基因的表达,而且出现翅芽外部畸形(外形改变、体积缩小)以及细胞凋亡。结论75μmol/LTSA调控某些重要的转录因子及有力地控制肌肉细胞的分化;高浓度TSA(≥750μmol/L)导致细胞凋亡和胚胎翅芽畸形;发育的鸡胚能够作为一个便利的、供体内研究药物(如HDAC抑制剂类等)的模型。
Objective To investigate the effect of trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, on the expression of some genes in the development of chicken wing embryos. To investigate the effect of TSA (75μmol / L, 750μmol / L, 1.5 mmol / L) on the development of chicken embryo buds, enhancing our ability to regulate chromatin remodeling and epigenetic gene expression, and our understanding of the anticancer drugs under development. Methods The embryo of chicken embryo was used as the experimental model, the wing buds were treated with TSA, the embryo of chicken was in situ hybridized, the expression of genes related to muscle development was detected, and the apoptosis status was detected by Nile blue sulfate . Results 75μmol / L TSA could inhibit the expression of MyoD and Myf5, a gene involved in muscle development, in chicken embryo buds. No apoptosis was observed in TSA (75μmol / L) -treated embryos. However, TSA (≥750μmol / L) not only strongly inhibited the expression of related genes, but also resulted in external deformities (shape change, volume reduction) and apoptosis in TSA. CONCLUSIONS: 75μmol / L TSA regulates some important transcription factors and strongly controls the differentiation of muscle cells. High concentrations of TSA (≥750μmol / L) lead to cell apoptosis and embryonic wing bud deformity. The developed chick embryo can be used as a convenient In vivo study of drugs (such as HDAC inhibitors, etc.) model.