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

Abstract

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

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]. .

References

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.

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

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