Han Jingyi, Li Jiaojiao, Liu Lianlian, Li Kaiyuan, Zhang Chun, Han Yong
Department of Physiology, Zunyi Medical University, Campus No.1 Road, Xinpu New District, Zunyi, 563006, Guizhou, China.
Sci Rep. 2025 Jan 17;15(1):2342. doi: 10.1038/s41598-025-85992-2.
In the vascular system, angiotensin II (Ang II) mediated vasoconstriction by inducing the production of 20-hydroxyeicosatetraenoic acid (20-HETE). However, the role of 20-HETE in Ang II-induced cardiac dysfunction had yet to be fully elucidated. This study investigated the effects of Ang II on CYP4A expression and 20-HETE production in H9c2 cells using RT-qPCR, Western blot, and ELISA. The role of 20-HETE in Ang II-induced cardiac hypertrophy was examined using DHE, MitoSOX, and JC-1 staining to evaluate reactive oxygen species (ROS) generation and mitochondrial membrane potential changes. The ERK/Akt and CaN/NFAT3 signaling pathways were analyzed through Western blot. Ang II was found to promote CYP4A expression and 20-HETE production in H9c2 cells via an AT1 receptor-dependent mechanism. Additionally, the upregulation of AT1 receptor expression by 20-HETE further confirms its facilitatory effect on the Ang II signaling pathway. Inhibition of 20-HETE synthesis or blockade of its receptor, G-protein-coupled receptor 75 (GPR75), significantly reversed Ang II-induced cardiac hypertrophy. This reversal was closely associated with 20-HETE-induced ROS production, oxidative stress, and activation of the Ca/CaN/NFAT3 signaling pathway. This study demonstrated that 20-HETE mediated Ang II-induced cardiac hypertrophy and, for the first time, highlighted the significant role of the GPR75 receptor in this process. These findings suggested that targeting 20-HETE reduction or blocking its receptor action could offer a novel therapeutic approach for cardiovascular diseases associated with Ang II.
在血管系统中，血管紧张素II（Ang II）通过诱导20-羟基二十碳四烯酸（20-HETE）的产生介导血管收缩。然而，20-HETE在Ang II诱导的心脏功能障碍中的作用尚未完全阐明。本研究使用RT-qPCR、蛋白质免疫印迹和酶联免疫吸附测定法研究了Ang II对H9c2细胞中CYP4A表达和20-HETE产生的影响。使用二氢乙锭（DHE）、线粒体超氧化物荧光探针（MitoSOX）和JC-1染色来评估活性氧（ROS）生成和线粒体膜电位变化，从而检测20-HETE在Ang II诱导的心肌肥大中的作用。通过蛋白质免疫印迹分析细胞外信号调节激酶/蛋白激酶B（ERK/Akt）和钙调神经磷酸酶/活化T细胞核因子3（CaN/NFAT3）信号通路。发现Ang II通过AT1受体依赖性机制促进H9c2细胞中CYP4A表达和20-HETE产生。此外，20-HETE对AT1受体表达的上调进一步证实了其对Ang II信号通路的促进作用。抑制20-HETE合成或阻断其受体G蛋白偶联受体75（GPR75）可显著逆转Ang II诱导的心肌肥大。这种逆转与20-HETE诱导的ROS产生、氧化应激以及Ca/CaN/NFAT3信号通路的激活密切相关。本研究表明，20-HETE介导Ang II诱导的心肌肥大，并且首次强调了GPR75受体在此过程中的重要作用。这些发现表明，靶向降低20-HETE或阻断其受体作用可能为与Ang II相关的心血管疾病提供一种新的治疗方法。
