DNA demethylation in the hypothalamus promotes transcription of Agtr1a and Slc12a2 and hypertension development.
DNA demethylation in the hypothalamus promotes transcription of Agtr1a and Slc12a2 and hypertension development.
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DOI：
10.1016/j.jbc.2023.105597
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发表时间：
2024-02
影响因子：
4.8
通讯作者：
Pan, Hui-Lin
中科院分区：
文献类型：
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作者：
Ghosh, Krishna;Zhou, Jing-Jing;Shao, Jian-Ying;Chen, Shao-Rui;Pan, Hui-Lin
下丘脑室旁核血管紧张素ⅡAT1a受体(Agtr1a编码)和Na+-K+-ClNKCC1(−共转运体1，编码SLc12a2)表达增加参与高血压的发生发展。然而，关于它们在高血压患者下丘脑室旁核中的转录调控却知之甚少。DNA甲基化是调控基因表达的重要表观遗传机制。在这里，我们确定了Agtr1a和SLc12a2的转录激活是否是自发性高血压大鼠(SHR)DNA甲基化改变的结果。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-京都大鼠(WKY)相比，SHR PVN中Agtr1a和Slc12a2启动子的CpG甲基化逐渐减少。染色质免疫沉淀-定量聚合酶链式反应显示，与WKY相比，SHR的DNA甲基转移酶(DNMT1和DNMT3A)和DNA甲基化阅读器蛋白2在Agtr1a和SLc12a2启动子上的浓集显著减少。相反，在PVN的Agtr1a和SLc12a2启动子上的10-11个易位酶(TET1-3)的丰度在SHR显著高于WKY。此外，向WKY的PVN内微量注射选择性DNMT抑制剂RG108可升高动脉血压，并相应增强PVN内Agtr1a和SLc12a2的mRNA水平。相反，在SHR的PVN内微量注射Tet特异性抑制剂C35可显著降低动脉血压，同时降低PVN内Agtr1a和SLc12a2的mRNA水平。综上所述，我们的发现表明，PVN中基因启动子上的DNMT/Tet开关导致的DNA低甲基化促进了Agtr1a和SLc12a2的转录，并促进了高血压的发生。
Increased expression of angiotensin II AT1A 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.