Benzoylaconitine is a natural product in the treatment of cardiovascular disease. However, its pharmacological effect, direct target protein, and molecular mechanisms for the treatment of heart failure are unclear. In this study, benzoylaconitine inhibited Ang II-induced cell hypertrophy and fibrosis in rat primary cardiomyocytes and rat fibroblasts, while attenuating cardiac function and cardiac remodeling in TAC mice. Using the limited proteolysis-mass spectrometry (LiP-MS) method, the angiotensin-converting enzyme 2 (ACE2) was confirmed as a direct binding target of benzoylaconitine for the treatment of heart failure. In ACE2-knockdown cells and ACE2 mice, benzoylaconitine failed to ameliorate cardiomyocyte hypertrophy, fibrosis, and heart failure. Online RNA-sequence analysis indicated p38/ERK-mediated mitochondrial reactive oxygen species (ROS) and nuclear factor kappa B (NF-κB) activation are the possible downstream molecular mechanisms for the effect of BAC-ACE2 interaction. Further studies in ACE2-knockdown cells and ACE2 mice suggested that benzoylaconitine targeted ACE2 to suppress p38/ERK-mediated mitochondrial ROS and NF-κB pathway activation. Our findings suggest that benzoylaconitine is a promising ACE2 agonist in regulating mitochondrial ROS release and inflammation activation to improve cardiac function in the treatment of heart failure.

中文翻译：

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苯甲酰乌头碱：一种有前途的 ACE2 靶向激动剂，可增强心力衰竭患者的心脏功能

苯甲酰乌头碱是治疗心血管疾病的天然产物。然而，其治疗心力衰竭的药理作用、直接靶蛋白和分子机制尚不清楚。在这项研究中，苯甲酰乌头碱抑制 Ang II 诱导的大鼠原代心肌细胞和大鼠成纤维细胞的细胞肥大和纤维化，同时减弱 TAC 小鼠的心脏功能和心脏重塑。使用有限蛋白水解质谱（LiP-MS）方法，证实血管紧张素转换酶2（ACE2）是苯甲酰乌头碱治疗心力衰竭的直接结合靶点。在 ACE2 敲低细胞和 ACE2 小鼠中，苯甲酰乌头碱未能改善心肌细胞肥大、纤维化和心力衰竭。在线 RNA 序列分析表明 p38/ERK 介导的线粒体活性氧 (ROS) 和核因子 kappa B (NF-κB) 激活是 BAC-ACE2 相互作用的可能下游分子机制。对 ACE2 敲低细胞和 ACE2 小鼠的进一步研究表明，苯甲酰乌头碱靶向 ACE2，抑制 p38/ERK 介导的线粒体 ROS 和 NF-κB 通路激活。我们的研究结果表明，苯甲酰乌头碱是一种很有前景的 ACE2 激动剂，可调节线粒体 ROS 释放和炎症激活，从而改善心力衰竭治疗中的心脏功能。