Hydralazine-induced promoter demethylation enhances sarcoplasmic reticulum Ca2+-ATPase and calcium homeostasis in cardiac myocytes
Hydralazine-induced promoter demethylation enhances sarcoplasmic reticulum Ca2+-ATPase and calcium homeostasis in cardiac myocytes
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DOI：
10.1038/labinvest.2011.92
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发表时间：
2011-09-01
影响因子：
5
通讯作者：
Chen, Yi-Jen
中科院分区：
文献类型：
--
作者：
Kao, Yu-Hsun;Cheng, Chen-Chuan;Chen, Yi-Jen
肌浆网（SR） Ca2+- atp酶（SERCA2a）在Ca2+稳态和心功能中起重要作用。SERCA2a的启动子区CpG岛含量高；因此，通过抑制甲基化的表观遗传修饰可以增强心肌细胞中SERCA2a的表达。Hydralazine是一种经常用于心力衰竭的药物，是一种潜在的DNA甲基化抑制剂。我们评估了肼是否可以通过调节甲基化增加SERCA2a表达来调节Ca2+处理。我们使用了吲哚-1荧光、实时RT-PCR、免疫印迹和甲基化特异性PCR来研究HL-1心肌细胞内Ca2+、RNA和蛋白质的表达以及SERCA2a的甲基化。（对照）给予羟嗪（1、10和30 μ M） 72小时后，羟嗪（10和30 μ M）增加了细胞内Ca2+瞬态和SR Ca2+含量。Hydralazine （10 μ M和30 μ M）降低SERCA2a启动子区域的甲基化，增加SERCA2a RNA和蛋白的表达。此外，10 μ M和30 μ M的肼嗪降低了DNA甲基转移酶1的表达。此外，在异丙肾上腺素诱导的心力衰竭大鼠中，用肼治疗可降低SERCA2a启动子甲基化，增加SERCA2a RNA表达。综上所述，肼嗪诱导的启动子去甲基化可能通过增加SERCA2a和调节心肌细胞钙稳态来改善心功能。实验室调查（2011）91,1291-1297；doi: 10.1038 / labinvest.2011.92;2011年7月11日在线发布
Sarcoplasmic reticulum (SR) Ca2+-ATPase (SERCA2a) plays an essential role in Ca2+ homeostasis and cardiac functions. The promoter region of SERCA2a has a high content of CpG islands; thus, epigenetic modification by inhibiting methylation can enhance SERCA2a expression in cardiomyocytes. Hydralazine, a drug frequently used in heart failure, is a potential DNA methylation inhibitor. We evaluated whether hydralazine can modulate Ca2+ handling through an increase in SERCA2a expression via regulating methylation. We used indo-1 fluorescence, real-time RT-PCR, immunoblotting, and methylation-specific PCR to investigate intracellular Ca2+, the expressions of RNA and protein, and methylation of SERCA2a in HL-1 cardiomyocytes with and without (control) the administration of hydralazine (1, 10, and 30 mu M) for 72 h. Hydralazine (10 and 30 mu M) increased the intracellular Ca2+ transients and SR Ca2+ contents. Hydralazine (10 and 30 mu M) decreased methylation in the SERCA2a promoter region and increased the RNA and protein expressions of SERCA2a. Additionally, hydralazine (10 and 30 mu M) decreased the expression of DNA methyltransferase 1. Moreover, treatment with hydralazine in isoproterenol-induced heart failure rats decreased the promoter methylation of SERCA2a and increased SERCA2a RNA expression. In conclusion, hydralazine-induced promoter demethylation may improve cardiac function through increasing SERCA2a and modulating calcium homeostasis in cardiomyocytes. Laboratory Investigation (2011) 91, 1291-1297; doi: 10.1038/labinvest.2011.92; published online 11 July 2011