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目的探讨表没食子儿茶素没食子酸酯[(-)-epigallo-catechin-3-gallate,EGCG]能否通过抑制组蛋白去乙酰化酶1(histone deacetylase 1,HDAC1)参与老年小鼠心脏SERCA2a的表达。方法24只16月龄雄性C57 BL/6老年小鼠随机数字表法分为EGCG+老年组[腹腔注射50 mg/(kg·d)的EGCG]、DMSO+老年组(腹腔注射与溶解EGCG同剂量的二甲基亚砜)以及未处理老年组,每组8只;3月龄雄性C57 BL/6成年小鼠作为青年组(n=8)。干预8周,收集心脏组织。RT-PCR检测Atp2a2及HDAC1的mRNA表达量,Western blot检测SERCA2a的表达量。Ch IP-Q-PCR检测Atp2a2启动子区组蛋白Ac H3K9水平及HDAC1、GATA4、Mef2c的结合量。结果 RT-PCR及Western blot结果表明,老年小鼠心脏SERCA2a在mRNA及蛋白水平的表达降低(P<0.05),HDAC1在mRNA水平表达升高(P<0.05)。Ch IP-Q-PCR结果表明,老年小鼠心脏Atp2a2启动子区HDAC1结合量升高,组蛋白Ac H3K9水平降低,GATA4、Mef2c的结合量降低(P<0.05)。EGCG干预8周后,老年小鼠心脏HDAC1的表达量及其在Atp2a2启动子区的结合量降低,组蛋白Ac H3K9水平升高,GATA4、Mef2c的结合量增加(P<0.05),EGCG在mRNA及蛋白水平升高了老年小鼠心脏SERCA2a的表达(P<0.05)。结论由HDAC1介导的组蛋白H3K9低乙酰化可能是老年小鼠心脏SERCA2a表达降低的关键机制之一,EGCG通过抑制这一过程升高老年小鼠心脏SERCA2a的表达。
Objective To investigate whether epigallo-catechin-3-gallate (EGCG) can participate in SERCA2a in the heart of old mice by inhibiting histone deacetylase 1 (HDAC1) expression. Methods Twenty-four male C57BL / 6 aged mice aged 16 months were randomly divided into EGCG + elderly group (intraperitoneal injection of EGCG at a dose of 50 mg / (kg · d)] and DMSO + elderly group (intraperitoneal injection and dissolution of EGCG at the same dose Dimethyl sulfoxide), and untreated elderly group, 8 in each group. Male C57BL / 6 mice at 3 months of age were used as youth group (n = 8). Intervention for 8 weeks, collecting heart tissue. The mRNA expression of Atp2a2 and HDAC1 was detected by RT-PCR and the expression of SERCA2a by Western blot. The level of histone Ac H3K9 and the binding of HDAC1, GATA4 and Mef2c in Atp2a2 promoter region were detected by ChIP-Q-PCR. Results The results of RT-PCR and Western blot showed that the expression of SERCA2a mRNA and protein in the heart of senile mice decreased (P <0.05) and the expression of HDAC1 increased at the mRNA level (P <0.05). Ch IP-Q-PCR results showed that the HDAC1 binding capacity of Atp2a2 promoter in the heart of aged mice was decreased, the level of histone Ac H3K9 was decreased and the binding capacity of GATA4 and Mef2c was decreased (P <0.05). At 8 weeks after EGCG treatment, the expression of HDAC1 and its binding to Atp2a2 promoter in the heart of aged mice increased, the levels of histone Ac H3K9, GATA4 and Mef2c increased (P <0.05) And protein levels increased the expression of SERCA2a in the heart of senile mice (P <0.05). Conclusions HDAC1-mediated hypo-acetylation of histone H3K9 may be one of the key mechanisms for the decrease of SERCA2a expression in the heart of senile mice. EGCG may up-regulate the expression of SERCA2a in aged mice by inhibiting this process.