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

中国畜牧兽医

• 生物技术 • 下一篇

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

阮秋燕，孙洪亮，蒙镇淼，刘晓淋，杜凤娇，石德顺，陆凤花

（广西亚热带农业生物资源保护与利用国家重点实验室，广西南宁 530005）

收稿日期:2013-11-04 出版日期:2014-05-20 发布日期:2014-06-25
通讯作者: 石德顺，研究员，主要从事动物胚胎工程与转基因动物研究。E-mail：ardsshi@gxu.edu.cn 陆凤花，研究员，主要从事动物胚胎工程与转基因动物研究。E-mail：lfhgggg@163.com
作者简介:阮秋燕（1989—），女，广西人，硕士生，研究方向：胚胎工程与技术。
基金资助:
国家“863”项目（2011AA100607）；国家自然科学基金项目（31072033）；教育部高等学校博士学科点博导基金项目（20114501110001）；广西杰出青年基金项目（2012GXNSFFA060004）；广西研究生教育创新计划资助项目（GXU11T31071）。

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

RUAN Qiu-yan, SUN Hong-liang, MENG Zhen-miao, LIU Xiao-lin, DU Feng-jiao, SHI De-shun，LU Feng-hua

(Guangxi Key Laboratory of Subtropical Bio-resource Conservation and Utilization, Nanning 530005,China)

Received:2013-11-04 Online:2014-05-20 Published:2014-06-25

摘要/Abstract

摘要： 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、750 nmol/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750 nmol/L 2个处理组细胞组蛋白乙酰化水平显著高于250 nmol/L处理组（P＜0.05），而500和750 nmol/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。

关键词: Scriptaid; 水牛胎儿成纤维细胞; 生长特性; 组蛋白乙酰化

Abstract: 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 characteristics, cell survival rate through trypan blue staining and the level of histone H3K18 acetylation (AcH3K18) by immunohistochemistry 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 significant difference on cell survival rate between Scriptaid-treated group and the control group (P＞0.05). In addition, immunohistochemistry detection found that the relative fluorescent intensity of AcH3K18 of BFF in the Scriptaid-treated group was significantly 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.

Key words:

Scriptaid; buffalo fetal fibroblasts; growth characteristic; histone acetylation

阮秋燕，孙洪亮，蒙镇淼，刘晓淋，杜凤娇，石德顺，陆凤花. 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[J]. 中国畜牧兽医.

RUAN Qiu-yan, SUN Hong-liang, MENG Zhen-miao, LIU Xiao-lin, DU Feng-jiao, SHI De-shun，LU Feng-hua. Effect of Different Concentrations of Scriptaid on the Development and Histone Acetylation Modification of Buffalo Fetal Fibroblasts[J]. .

参考文献

1 王雷, 丁春华, 栾爽艳. 我国水牛奶业的发展现状与开发前景［J］.中国奶牛，2008，4:8～11. 2 李正阳. Scriptaid提高小鼠体细胞核移植克隆胚早期发育能力的实验研究［D］. 广州：南方医科大学, 2012. 3 陆凤花, 石德顺, 谢英，等.不同来源的供体细胞对水牛体细胞核移植效果的影响［J］. 广西农业生物科学，2005,25(2):89～93. 4 高川, 熊显荣, 符梅，等. Scriptaid处理牦牛成纤维细胞对组蛋白乙酰化及重编程的影响［J］. 农业生物技术学报，2013,21(1):62～68. 5 温兵强. Scriptaid和咖啡因对绵羊体细胞核移植效果的影响研究［D］. 保定：河北农业大学, 2012. 6 Akagi S, Matsukawa K, Mizutani E, et al. Treatment with a histone deacetylase inhibitor after nuclear transfer improves the preimplantation development of cloned bovine embryos［J］. J Reprod Dev, 2011, 57(1):120～126. 7 Bourc’His D, Le Bourhis D, Patin D, et al. Delayed and incomplete reprogramming of chromosome methylation patterns in bovine cloned embryos［J］. Curr Biol, 2001,11(19):1542～1546. 8 Dean W, Santos F, Stojkovic M, et al. Conservation of methylation reprogramming in mammalian development: Aberrant reprogramming in cloned embryos［J］. Proc Natl Acad Sci USA, 2001, 98(24):13734～13738. 9 Huang J C, Lei Z L, Shi L H, et al. Comparison of histone modifications in in vivo and in vitro fertilization mouse embryos［J］. Biochem Biophys Res Commun, 2007, 354(1):77～83. 10 Iager A E, Ragina N P, Ross P J, et al. Trichostatin A improves histone acetylation in bovine somatic cell nuclear transfer early embryos［J］. Cloning Stem Cells, 2008, 10(3):371～379. 11 Su G H, Sohn T A, Ryu B, et al. A novel histone deacetylase inhibitor identified by high-throughput transcriptional screening of a compound library［J］. Cancer Res, 2000, 60(12):3137～3142. 12 Turner B M. Histone acetylation as an epigenetic determinant of long-term transcriptional competence［J］. Cell Mol Life Sci, 1998, 54(1):21～31. 13 Van Thuan N, Bui H T, Kim J H, et al. The histone deacetylase inhibitor scriptaid enhances nascent mRNA production and rescues full-term development in cloned inbred mice［J］. Reproduction, 2009, 138(2):309～317. 14 Zhao J, Ross J W, Hao Y, et al. Significant improvement in cloning efficiency of an inbred miniature pig by histone deacetylase inhibitor treatment after somatic cell nuclear transfer［J］. Biol Reprod, 2009, 81(3):525～530. 15 Zlatanova J, Caiafa P, Van Holde K. Linker histone binding and displacement: Versatile mechanism for transcriptional regulation［J］. Faseb J, 2000, 14(12):1697～1704.

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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

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