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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 investigate the effects of prenatal alcohol exposure on gene expression of CBP / P300-interacting transactivator with EDRI 2 and CITED2 in mouse embryos and young mice, and to explore the relationship between alcohol exposure during pregnancy and CITED2 promoter Cp G island methylation level of the relationship. Methods: Twenty pregnant rats [embryonic mouse developmental period (E0.5-E20.5)] were inoculated intragastrically with 10μl / (g · d) and 56% drinking liquor as the intervention group, and saline Pregnant rats as control group. Cardiac tissues from E14.5, E17.5, neonatal rat hearts and 7 days after birth were used as the specimens. Expression of CITED2, GATA binding protein 4 (GATA4) mRNA was detected by q RT-PCR The changes of CpG island methylation in CITED2 promoter region were detected by MSP and BSP. Results: Compared with the control group [(0.819 ± 0.176), (1.115 ± 0.118)], the GATA4 gene expression in heart during E14.5 and E17.5 [(1.538 ± 0.071) and (1.734 ± 0.101) The expression levels were increased by 1.88 and 1.56 folds in the control group (P <0.05). CITED2 expression in E14.5, E17.5 and newborn mice (0.896 ± 0.069,1.336 ± 0.121,0.988 ± 0.108) compared with the control group (1.246 ± 0.158,1.833 ± 0.086,1.355 ± 0.126) expression decreased, respectively (P <0.05). The expression of CITED2 mRNA in the alcohol exposure group (1.276 ± 0.097) was lower than that in the normal group (1.024 ± 0.078) at 7 days after the alcohol exposure, but the difference was not statistically significant (P > 0.05). CONCLUSION: Alcohol exposure during pregnancy plays a negative regulatory role on the expression of m RNA in CITED2 gene, indicating that alcohol exposure during pregnancy may inhibit the expression of CITED2 gene in some way. Alcohol exposure during pregnancy may not affect the CITED2 gene expression by changing the methylation level of CpG island in the promoter region of CITED2 gene, thus affecting cardiac development.