不同浓度的组蛋白去乙酰化酶抑制剂Scriptaid对水牛胎儿成纤维细胞生长及其组蛋白乙酰化修饰的影响

Effect of Different Concentrations of Scriptaid on the Development and Histone Acetylation Modification of Buffalo Fetal Fibroblasts

研究旨在探讨组蛋白去乙酰化酶抑制剂Scriptaid对水牛胎儿成纤维细胞(BFF)生长特性、细胞毒性及其组蛋白H3K18乙酰化修饰的影响,为今后研究供体细胞的重编程及提高水牛核移植效率提供一定的理论依据。试验采用不同浓度(0、250、500、750nmol/L)的Scriptaid分别处理BFF后观察细胞的生长形态,然后利用台盼蓝染色法检测细胞的存活率,同时通过细胞免疫组化技术分析细胞组蛋白H3K18乙酰化的表达情况。结果发现,BFF经不同浓度的Scriptaid处理后,细胞形态没有显著变化,为长梭形、立体感较强,生长曲线与对照组相似,均呈"S"型分布,且各处理组细胞的存活率与对照组之间差异不显著(P>0.05);此外,Scriptaid处理组的细胞组蛋白H3K18乙酰化的相对荧光强度显著高于对照组(P<0.05),其中500和750nmol/L 2个处理组细胞组蛋白乙酰化水平显著高于250nmol/L处理组(P<0.05),而500和750nmol/L 2个处理组之间差异不显著(P>0.05)。可见,适合浓度的Scriptaid对BFF细胞形态和存活率影响不大,且能显著提高BFF的组蛋白乙酰化水平。

The purpose of this study was to investigate the effect of various concentrations of histone deacetylase inhibitor （Scriptaid） on the growth characteristics, cytotoxicity and the level of histone H3K18 acetylation （AcH3K18） of buffalo fetal fibroblasts （BFF）, so that to find a novel way to improve the efficiency of buffalo somatic cell nuclear transfer in future. The experiment investigated the effect of various concentrations （0, 250, 500, 750 nmol/L） of Scriptaid on the growth characteris- tics, cell survival rate through trypan blue staining and the level of histone H3K18 acetylation （AcH3K18） by immunohisto- chemistry detection. The results showed that there was no effect on the biology characteristics of the BFF treated with Scriptaid, the growth curve was similar with that of the control group, showing ＂S＂ distribution, moreover, there was no sig- nificant difference on cell survival rate between Scriptaid-treated group and the control group （P〉0.05）. In addition, immuno- histochemistry detection found that the relative fluorescent intensity of AcH3K18 of BFF in the Scriptaid-treated group was sig- nificantly higher than that of the non-treated group （P〈0.05）, and the levels of AcH3K18 of BFF in the 500 and 750 nmol/L groups were significantly improved compared with that of the 250 nmol/L group （P〈0.05）, but there was no significant difference between 500 and 750 nmol/L treatment groups （P〉0.05）. These results indicated that an optimum concentration of Scriptaid had little effect on the cell morphology and cell survival rate of BFF, and could significantly improve the levels of histone acetylation.