Differential regulation of DNA methylation versus histone acetylation in cardiomyocytes during HHcy in vitro and in vivo: an epigenetic mechanism
Differential regulation of DNA methylation versus histone acetylation in cardiomyocytes during HHcy in vitro and in vivo: an epigenetic mechanism
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DOI：
10.1152/physiolgenomics.00168.2013
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发表时间：
2014-04-01
影响因子：
4.6
通讯作者：
Tyagi, Suresh C.
中科院分区：
文献类型：
--
作者：
Chaturvedi, Pankaj;Kalani, Anuradha;Tyagi, Suresh C.
体外和体内高同型半胱氨酸诱导心肌细胞DNA甲基化与组蛋白乙酰化的差异调节：一种表观遗传学机制。Physiol Genomics 46：245-255，2014.首次发表于2014年2月4日; doi：10.1152/physiolgenomics.00168.2013.-同型半胱氨酸介导的心脏威胁的机制知之甚少。同型半胱氨酸是S-腺苷甲硫氨酸（甲基供体）通过甲硫氨酸的前体，间接参与DNA、RNA和蛋白质的甲基化现象。我们以前报道过，心脏特异性缺失N-甲基-D-天冬氨酸受体1（NMDAR 1）可改善同型半胱氨酸引起的心脏威胁，在这项研究中，我们的目的是探讨NMDAR 1在心力衰竭的表观遗传机制中的作用，使用心肌细胞在高同型半胱氨酸（HHcy）。高同型半胱氨酸水平激活NMDAR 1，从而通过调节心肌细胞中的DNA甲基转移酶1（DNMT 1）、HDAC 1、miRNA和MMP 9，导致DNA甲基化相对于组蛋白乙酰化异常。用100 μ M同型半胱氨酸以剂量依赖性方式处理在Claycomb培养基中培养的HL-1心肌细胞。在不存在和存在10 μ M高半胱氨酸的情况下以剂量依赖性方式加入NMDAR 1拮抗剂（MK 801）。通过实时PCR和蛋白质印迹法评估DNMT 1、组蛋白脱乙酰酶1（HDAC 1）、NMDAR 1、microRNA（miR）-133 a和miR- 499的表达。通过检查5=-甲基胞嘧啶DNA甲基化和染色质免疫沉淀来确定甲基化和乙酰化水平。使用高同型半胱氨酸血症小鼠模型（CBS +/-）来确认体内结果。在HHcy中，NMDAR 1、DNMT 1和基质金属蛋白酶9的表达随着H3 K9乙酰化的增加而增加，而HDAC 1、miR-133 a和miR- 499在心肌细胞中的表达减少。在CBS +/-小鼠的心脏组织中获得了类似的结果。高同型半胱氨酸水平通过NMDAR 1、miR-133 a、miR- 499和DNMT 1引发心血管重塑。HDAC 1的减少和H3 K9乙酰化和DNA甲基化的增加提示HHcy中的染色质重塑。
Differential regulation of DNA methylation versus histone acetylation in cardiomyocytes during HHcy in vitro and in vivo: an epigenetic mechanism. Physiol Genomics 46: 245-255, 2014. First published February 4, 2014; doi:10.1152/physiolgenomics.00168.2013.- The mechanisms of homocysteine-mediated cardiac threats are poorly understood. Homocysteine, being the precursor to S-adenosyl methionine (a methyl donor) through methionine, is indirectly involved in methylation phenomena for DNA, RNA, and protein. We reported previously that cardiac-specific deletion of N-methyl-D-aspartate receptor-1 (NMDAR1) ameliorates homocysteine-posed cardiac threats, and in this study, we aim to explore the role of NMDAR1 in epigenetic mechanisms of heart failure, using cardiomyocytes during hyperhomocysteinemia (HHcy). High homocysteine levels activate NMDAR1, which consequently leads to abnormal DNA methylation vs. histone acetylation through modulation of DNA methyltransferase 1 (DNMT1), HDAC1, miRNAs, and MMP9 in cardiomyocytes. HL-1 cardiomyocytes cultured in Claycomb media were treated with 100 mu M homocysteine in a dose-dependent manner. NMDAR1 antagonist (MK801) was added in the absence and presence of homocysteine at 10 mu M in a dose-dependent manner. The expression of DNMT1, histone deacetylase 1 (HDAC1), NMDAR1, microRNA (miR)- 133a, and miR- 499 was assessed by real-time PCR as well as Western blotting. Methylation and acetylation levels were determined by checking 5=- methylcytosine DNA methylation and chromatin immunoprecipitation. Hyper-homocysteinemic mouse models (CBS +/-) were used to confirm the results in vivo. In HHcy, the expression of NMDAR1, DNMT1, and matrix metalloproteinase 9 increased with increase in H3K9 acetylation, while HDAC1, miR- 133a, and miR- 499 decreased in cardiomyocytes. Similar results were obtained in heart tissue of CBS +/- mouse. High homocysteine levels instigate cardiovascular remodeling through NMDAR1, miR- 133a, miR- 499, and DNMT1. A decrease in HDAC1 and an increase in H3K9 acetylation and DNA methylation are suggestive of chromatin remodeling in HHcy.