摘要
目的:研究孕期酒精暴露对小鼠胚胎及幼鼠心脏CBP/P300反式作用因子2(CBP/P300-interacting transactivator with EDrich 2,CITED2)基因表达的影响,探讨孕期酒精暴露与CITED2基因启动子区Cp G岛甲基化水平的关系。方法:以10μl/(g·d),56%的饮用白酒给20只孕鼠[胚胎小鼠发育期(E0.5-E20.5)]连续灌胃作为干预组,生理盐水灌胃20只孕鼠作为对照组。取E14.5、E17.5、新生鼠心脏及生后7 d小鼠心脏组织作为标本,q RT-PCR检测心脏发育相关基因CITED2、GATA4结合蛋白4(GATA binding protein 4,GATA4)m RNA表达量的变化,MSP及BSP检测CITED2基因启动子区Cp G岛甲基化变化情况。结果:孕期酒精暴露致小鼠心脏发育相关基因GATA4在E14.5及E17.5[(1.538±0.071)、(1.734±0.101)]较对照组[(0.819±0.176)、(1.115±0.118)]表达水平升高,分别为其对照组1.88、1.56倍(P<0.05)。CITED2的表达在E14.5、E17.5及新生小鼠(0.896±0.069,1.336±0.121,0.988±0.108)较对照组(1.246±0.158,1.833±0.086,1.355±0.126)表达水平降低,分别为其对照组0.72、0.73、0.74倍(P<0.05),酒精暴露组生后7 d CITED2 m RNA表达量(1.276±0.097)较正常组(1.024±0.078)降低,但差异无统计学意义(P>0.05)。结论:孕期酒精暴露对CITED2基因m RNA表达水平起到了负调节作用,说明孕期酒精暴露通过了某种途径对CITED2基因表达起到了抑制作用。孕期酒精暴露可能并不是通过改变CITED2基因启动子区Cp G岛甲基化水平来影响CITED2基因的表达,从而影响心脏发育。
Objective :To reveal the effect of prenatal alcohol exposure on CBP/P300-interacting transactivator with ED-rich 2 (CIT- ED2) gene expression in mouse embryos and heart,and to investigate the relationship between alcohol exposure during pregnancy and the methylation level of CpG island in the promoter region of CITED2 gene. Methods:Twenty pregnant mice were exposed to a single dose of alcohol(10 μl/g/d, 56% alcohol) by garage between EO.5-E20.5;another 20 pregnant mice exposed to saline lavage were taken as control group. Embryonic hearts of E14.5 ,E17.5 fetal mice, newborn mice and postnatal 7 d mice were collected re- spectively. The mRNA expressions of the heart development-related genes (CITED2, GATA binding protein 4(GATA4)) were mea- sured by Q-PCR method. CpG islangds, methylation of CITED2 promoter was measured, Results:Alcohol exposure increased the ex- pression of GATA4 at E14.5 and E17.5 by 1.88 and 1.56 folds,respectively(P〈0.05),decreased the expression of CITED2 at E14.5, E17.5 and newborn by 0.72,0.73 and 0.74 fold(P〈0.05). Alcohol exposure increased the expression of CITED2 on postnatal 7 d,however it showed no statistical difference(P〉05). Conclusion: Alcohol exposure during pregnancy exerts a negative regulatory effect on CITED2 gene expression,the mechanism of which may not through changing methylation of CpG islands of CIT- ED2 thus affecting cardiac development.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2016年第7期669-674,共6页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(编号:81270234)
重庆市卫生局医学科研计划资助项目(编号:2009-2-259)
重庆市渝中区科技计划资助项目(编号:20120216)