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目的明确GATA4在小鼠胚胎心脏发育过程中的时序表达,并阐明其组蛋白修饰调控机制。方法选取胎龄11.5 d(E11.5)至新生0.5 d胎鼠心脏,RT-PCR检测胎鼠心肌细胞中GATA4mRNA表达水平,CHIP-QPCR检测GATA4启动子区组蛋白H3K9ac水平及与CBP/P300的结合水平。用CBP30干预心肌祖细胞,RT-PCR检测心肌祖细胞在不同浓度CBP30(0.5、1、2、4μmol/L)干预不同时间点(6、12、24、36 h)后GATA4 mRNA表达水平,CHIP-QPCR检测心肌祖细胞中GATA4启动子区组蛋白H3K9ac水平及与CBP/P300的结合水平。结果 1小鼠胚胎心脏发育过程中GATA4表达量,E14.5明显高于E11.5(P<0.05);CHIP结果显示E14.5GATA4启动子区组蛋白H3K9ac水平高于E11.5(P<0.05);E14.5与CBP/P300的结合水平亦高于E11.5(P<0.05)。2CBP30干预心肌祖细胞后,GATA4表达量较对照组明显降低(P<0.05)。CHIP结果显示CBP30组GATA4启动子区组蛋白H3K9ac水平及与CBP/P300的结合水平较对照组显著降低(P<0.05)。结论 CBP/P300可通过介导组蛋白H3K9乙酰化修饰参与调控GATA4在小鼠胚胎心脏发育过程中的时序表达。
Objective To determine the temporal expression of GATA4 in mouse embryonic heart development and elucidate the regulatory mechanism of histone modification. Methods The heart of fetal rat was collected from 11.5 days gestation (E11.5) to 0.5 days gestation. The expression of GATA4 mRNA in fetal rat cardiomyocytes was detected by RT-PCR. The level of histone H3K9ac in GATA4 promoter region was detected by CHIP-QPCR, Binding level. The cardiac progenitor cells were treated with CBP30, and the expression of GATA4 mRNA was detected by RT-PCR after different concentrations of CBP30 (0.5, 1, 2, 4μmol / L) for different time points (6, 12, 24, 36 h) The levels of histone H3K9ac in GATA4 promoter region and the level of binding to CBP / P300 in cardiac progenitor cells were detected by qPCR. Results The expression level of GATA4 in E14.5 was significantly higher than that in E11.5 (P <0.05) during embryonic heart development. The level of H3K9ac in E14.5GATA4 promoter region was higher than that in E11.5 by CHIP ). The binding level of E14.5 to CBP / P300 was also higher than E11.5 (P <0.05). After intervention of 2CBP30 on cardiac progenitor cells, the expression of GATA4 was significantly lower than that of the control group (P <0.05). CHIP results showed that the levels of H3K9ac and the binding to CBP / P300 in GATA4 promoter region of CBP30 group were significantly lower than those in control group (P <0.05). Conclusion CBP / P300 may be involved in the regulation of GATA4 expression in mouse embryonic heart through the acetylation of histone H3K9.