Alcohol Consumption During Gestation Causes Histone3 Lysine9 Hyperacetylation and an Alternation of Expression of Heart Development-Related Genes in Mice
Alcohol Consumption During Gestation Causes Histone3 Lysine9 Hyperacetylation and an Alternation of Expression of Heart Development-Related Genes in Mice
复制标题
妊娠期间饮酒会导致小鼠组蛋白 3 赖氨酸 9 过度乙酰化以及心脏发育相关基因表达的改变
DOI：
10.1111/acer.12518
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发表时间：
2014-09-01
影响因子：
3.2
通讯作者：
Tian, Jie
中科院分区：
文献类型：
--
作者：
Pan, Bo;Zhu, Jing;Tian, Jie
背景：妊娠期酒精滥用可能导致先天性心脏病（CHD）。酒精性心脏畸形的机制尚不清楚。最近的研究表明，组蛋白修饰可能在这一病理过程中起着至关重要的作用。此外，我们以前的研究报告，乙醇可以诱导组蛋白3赖氨酸9（H3 K9）的高乙酰化和心脏发育相关基因的表达在体外。本研究的目的是探讨妊娠期饮酒对H3 K9乙酰化失衡和心脏发育相关基因表达的影响。采用苏木精-伊红（H&E）染色观察胚胎心脏的结构。Western blotting和实时定量PCR检测H3 K9乙酰化水平和基因表达。结果：心脏组织H&E染色显示E17.5胚胎心脏异常，HAT和HDAC活性均高于正常胚胎。H3 K9乙酰化水平在E17.5时达到高峰，出生时急剧下降至较低水平，此后一直维持在较低水平。酒精暴露分别在E11.5、E14.5、E17.5和E18.5增加H3 K9乙酰化（p < 0.05），并在E14.5和E17.5增加胚胎心脏中Gata 4的表达，在E14.5增加Mef 2c的表达，在E14.5和E17.5增加Nkx2.5的表达（p < 0.05），但Tbx 5没有（p > 0.05）。结论：胚胎发育过程中H3 K9乙酰化水平的变化存在一定的时间过程，表明子宫内酒精暴露可诱导HAT活性的增加，从而导致H3 K9乙酰化水平的升高和心脏发育相关基因表达的增加。这些发现揭示了一种新的表观遗传机制，将妊娠期间的饮酒与胎儿CHD的发展联系起来。
Background: Alcohol abuse during gestation may cause congenital heart diseases (CHDs). The underlying mechanisms of alcohol-induced cardiac deformities are still not clear. Recent studies suggest that histone modification may play a crucial role in this pathological process. Moreover, our previous studies reported that ethanol could induce histone3 lysine9 (H3K9) hyperacetylation and overexpression of heart development-related genes in vitro. The aim of this study was to investigate the effect of alcohol consumption during gestation on the imbalance of H3K9 acetylation and the alternation of the expression of heart development-related genes during cardiogenesis.Methods: Pregnant mice were exposed to a single dose of alcohol (10 mu l/g/d, 56% alcohol) by gavage every day in the morning from embryo day 7.5 (E7.5) to E15.5. Hematoxylin and eosin (H&E) staining was applied for observing the structure of the embryonic hearts. Western blotting and quantitative real-time polymerase chain reaction were used for detecting the level of H3K9 acetylation and gene expression. Histone acetyltransferase (HAT) and histone deacetylase (HDAC) activities were detected by colorimetric assay and fluorometric assay.Results: H&E staining of cardiac tissue showed abnormalities of embryonic hearts at E17.5. The level of H3K9 acetylation reached peak at E17.5 and decreased sharply to a low level at birth and maintained at low level afterward. Alcohol exposure increased H3K9 acetylation at E11.5, E14.5, E17.5, and E18.5, respectively (p < 0.05), and enhanced the expression of Gata4 in the embryonic hearts at E14.5 and E17.5, Mef2c at E14.5, and Nkx2.5 at E14.5 and E17.5, (p < 0.05) but not for Tbx5 (p > 0.05). On embryonic day 17.5, HAT activities of embryonic hearts increased significantly, however alcohol exposure did not alter HDAC activities.Conclusions: These data indicate a time course of H3K9 acetylation change during heart development and demonstrate that alcohol exposure in utero may induce an increase of HAT activities, which results in H3K9 hyperacetylation and an increase of the expression of heart development-related genes. These findings reveal a novel epigenetic mechanism that connects the alcohol consumption during the pregnancy and the development of CHD in the fetus.