Center for Neuroscience and Pain Research, Department of Anesthesiology and Perioperative Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
Center for Neuroscience and Pain Research, Department of Anesthesiology and Perioperative Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
J Biol Chem. 2024 Feb;300(2):105597. doi: 10.1016/j.jbc.2023.105597. Epub 2023 Dec 29.
Increased expression of angiotensin II AT receptor (encoded by Agtr1a) and Na-K-Cl cotransporter-1 (NKCC1, encoded by Slc12a2) in the hypothalamic paraventricular nucleus (PVN) contributes to hypertension development. However, little is known about their transcriptional control in the PVN in hypertension. DNA methylation is a critical epigenetic mechanism that regulates gene expression. Here, we determined whether transcriptional activation of Agtr1a and Slc12a2 results from altered DNA methylation in spontaneously hypertensive rats (SHR). Methylated DNA immunoprecipitation and bisulfite sequencing-PCR showed that CpG methylation at Agtr1a and Slc12a2 promoters in the PVN was progressively diminished in SHR compared with normotensive Wistar-Kyoto rats (WKY). Chromatin immunoprecipitation-quantitative PCR revealed that enrichment of DNA methyltransferases (DNMT1 and DNMT3A) and methyl-CpG binding protein 2, a DNA methylation reader protein, at Agtr1a and Slc12a2 promoters in the PVN was profoundly reduced in SHR compared with WKY. By contrast, the abundance of ten-eleven translocation enzymes (TET1-3) at Agtr1a and Slc12a2 promoters in the PVN was much greater in SHR than in WKY. Furthermore, microinjecting of RG108, a selective DNMT inhibitor, into the PVN of WKY increased arterial blood pressure and correspondingly potentiated Agtr1a and Slc12a2 mRNA levels in the PVN. Conversely, microinjection of C35, a specific TET inhibitor, into the PVN of SHR markedly reduced arterial blood pressure, accompanied by a decrease in Agtr1a and Slc12a2 mRNA levels in the PVN. Collectively, our findings suggest that DNA hypomethylation resulting from the DNMT/TET switch at gene promoters in the PVN promotes transcription of Agtr1a and Slc12a2 and hypertension development.
血管紧张素 II AT 受体（由 Agtr1a 编码）和钠-钾-氯共转运蛋白-1（NKCC1，由 Slc12a2 编码）在下丘脑室旁核（PVN）中的表达增加导致高血压的发展。然而，关于高血压患者 PVN 中它们的转录控制知之甚少。DNA 甲基化是一种关键的表观遗传机制，可调节基因表达。在这里，我们确定血管紧张素 II AT 受体（Agtr1a）和钠-钾-氯共转运蛋白-1（Slc12a2）的转录激活是否是由于自发性高血压大鼠（SHR）中 DNA 甲基化的改变所致。甲基化 DNA 免疫沉淀和亚硫酸氢盐测序-PCR 显示，与正常血压的 Wistar-Kyoto 大鼠（WKY）相比，SHR 中 PVN 中 Agtr1a 和 Slc12a2 启动子的 CpG 甲基化逐渐减少。染色质免疫沉淀-定量 PCR 显示，SHR 中 PVN 中 DNA 甲基转移酶（DNMT1 和 DNMT3A）和 DNA 甲基化阅读器蛋白甲基-CpG 结合蛋白 2 与 Agtr1a 和 Slc12a2 启动子的结合明显减少。相比之下，SHR 中 Agtr1a 和 Slc12a2 启动子处的 ten-eleven 易位酶（TET1-3）的丰度远高于 WKY。此外，将选择性 DNMT 抑制剂 RG108 微注射到 WKY 的 PVN 中会增加动脉血压，并相应地增强 PVN 中 Agtr1a 和 Slc12a2 的 mRNA 水平。相反，将特定的 TET 抑制剂 C35 微注射到 SHR 的 PVN 中会显著降低动脉血压，并伴有 PVN 中 Agtr1a 和 Slc12a2 的 mRNA 水平降低。总之，我们的研究结果表明，PVN 中基因启动子处的 DNMT/TET 开关导致的 DNA 低甲基化促进了 Agtr1a 和 Slc12a2 的转录和高血压的发展。
