Kohata Nao, Kurihara Isao, Yokota Kenichi, Kobayashi Sakiko, Murai-Takeda Ayano, Mitsuishi Yuko, Nakamura Toshifumi, Morisaki Mitsuha, Kozuma Takahide, Torimitsu Takuto, Kawai Miki, Itoh Hiroshi
Department of Internal Medicine, Keio University School of Medicine, Tokyo, Japan.
Department of Medical Education, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama, 359-8513, Japan.
Hypertens Res. 2022 Apr;45(4):641-649. doi: 10.1038/s41440-022-00859-7. Epub 2022 Feb 17.
Mineralocorticoid receptor (MR) and its ligand aldosterone play a central role in controlling blood pressure by promoting sodium reabsorption in the kidney. Coregulators are recruited to regulate the activation of steroid hormone receptors. In our previous study, we identified several new candidates for MR coregulators through liquid chromatography-tandem mass spectrometry analysis using a biochemical approach. Lysine-specific demethylase 1 (LSD1) was identified as a candidate. The relationship between LSD1 and salt-sensitive hypertension has been reported; however, the role of MR in this condition is largely unknown. Here, we investigated the functions of LSD1 as a coregulator of MR. First, a coimmunoprecipitation assay using HEK293F cells showed specific interactions between MR and LSD1. A chromatin immunoprecipitation study demonstrated LSD1 recruitment to the gene promoter of epithelial Na channel (ENaC), a target gene of MR. Reduced LSD1 expression by treatment with shRNA potentiated the hormonal activation of ENaC and serum/glucocorticoid-regulated kinase 1, another target gene of MR, indicating that LSD1 is a corepressor of MR. In an animal study, mice with kidney-specific LSD1 knockout (LSD1KSP-Cre mice) developed hypertension after a high-salt diet without elevation of aldosterone levels, which was counteracted by cotreatment with spironolactone, an MR antagonist. In conclusion, our in vitro and in vivo studies demonstrated that LSD1 is a newly identified corepressor of MR.
盐皮质激素受体（MR）及其配体醛固酮通过促进肾脏对钠的重吸收在控制血压方面发挥核心作用。共调节因子被招募来调节类固醇激素受体的激活。在我们之前的研究中，我们使用生化方法通过液相色谱 - 串联质谱分析鉴定了几种新的MR共调节因子候选物。赖氨酸特异性去甲基化酶1（LSD1）被鉴定为候选物。LSD1与盐敏感性高血压之间的关系已有报道；然而，MR在这种情况下的作用在很大程度上尚不清楚。在这里，我们研究了LSD1作为MR共调节因子的功能。首先，使用HEK293F细胞进行的免疫共沉淀试验显示MR和LSD1之间存在特异性相互作用。染色质免疫沉淀研究表明LSD1被招募到上皮钠通道（ENaC）的基因启动子，ENaC是MR的一个靶基因。用shRNA处理降低LSD1表达增强了ENaC和血清/糖皮质激素调节激酶1（MR的另一个靶基因）的激素激活，表明LSD1是MR的一个共抑制因子。在一项动物研究中，肾脏特异性LSD1基因敲除小鼠（LSD1KSP - Cre小鼠）在高盐饮食后出现高血压，而醛固酮水平没有升高，螺内酯（一种MR拮抗剂）联合治疗可抵消这种高血压。总之，我们的体外和体内研究表明LSD1是一种新鉴定的MR共抑制因子。
