Liu Yan-Dong, Qiao Wei, Pei Zhao-Hui, Song Guo-Liang, Jin Wei, Zhong Wei-Bing, Deng Qin-Qin
the Second Department of Cardiovascular Disease, Nanchang People's Hospital (Nanchang Third Hospital) Nanchang 330009, China.
Health Institute of Nanchang Normal University Nanchang 330009, China.
Zhongguo Zhong Yao Za Zhi. 2025 Aug;50(16):4679-4689. doi: 10.19540/j.cnki.cjcmm.20250512.503.
This study aims to explore the specific mechanism by which puerarin inhibits ferroptosis and improves the myocardial contractile function in myocardial hypertrophy through the nuclear factor erythroid 2-related factor 2(Nrf2)/antioxidant response element(ARE)/heme oxygenase-1(HO-1) signaling pathway. The hypertrophic cardiomyocyte model was established using phenylephrine, and H9c2 cells were divided into control group, model group, puerarin group, and puerarin+ML385 group. Cell viability and surface area were detected by cell counting kit-8(CCK-8) and immunofluorescence experiments. The mitochondrial membrane potential and Ca(2+) concentration were measured. The ferroptosis-related indicators were detected by biochemical and fluorescence staining methods. The expression of proteins related to ferroptosis and the Nrf2/ARE/HO-1 signaling pathway was detected by Western blot. A myocardial hypertrophy model was established, and 40 rats were randomly divided into sham group, model group, puerarin group, and puerarin+Nrf2 inhibitor(ML385) group, with 10 rats in each group. Echocardiogram, hemodynamic parameters, and myocardial hypertrophy parameters were measured. Histopathological changes of myocardial tissues were observed by hematoxylin and eosin(HE) staining and Masson staining. Biochemical methods, enzyme-linked immunosorbent assay(ELISA), and fluorescence staining were used to detect inflammatory factors and ferroptosis-related indicators. Immunohistochemistry was used to detect the expression of proteins related to ferroptosis and the Nrf2/ARE/HO-1 signaling pathway. Cell experiments showed that puerarin intervention significantly enhanced the viability of hypertrophic cardiomyocytes, reduced their surface area, and restored mitochondrial membrane potential and Ca(2+) homeostasis. Mechanism studies revealed that puerarin promoted Nrf2 nuclear translocation, upregulated the expression of HO-1, solute carrier family 7 member 11(SLC7A11), and glutathione peroxidase 4(GPX4), and decreased malondialdehyde(MDA), reactive oxygen species(ROS), and iron levels. These protective effects were reversed by ML385. In animal experiments, puerarin improved cardiac function in rats with myocardial hypertrophy, alleviated myocardial hypertrophy and fibrosis, inhibited inflammatory responses and ferroptosis, and promoted nuclear Nrf2 translocation and HO-1 expression. However, combined intervention with ML385 led to deterioration of hemodynamics and a rebound in ferroptosis marker levels. In conclusion, puerarin may inhibit cardiomyocyte ferroptosis through the Nrf2/ARE/HO-1 signaling pathway, thereby improving myocardial contractile function in myocardial hypertrophy.
本研究旨在探讨葛根素通过核因子红细胞2相关因子2（Nrf2）/抗氧化反应元件（ARE）/血红素加氧酶-1（HO-1）信号通路抑制铁死亡并改善心肌肥厚中心肌收缩功能的具体机制。采用去甲肾上腺素建立肥厚型心肌细胞模型，将H9c2细胞分为对照组、模型组、葛根素组和葛根素+ML385组。通过细胞计数试剂盒-8（CCK-8）和免疫荧光实验检测细胞活力和表面积。测量线粒体膜电位和Ca²⁺浓度。采用生化和荧光染色方法检测铁死亡相关指标。通过蛋白质免疫印迹法检测铁死亡相关蛋白以及Nrf2/ARE/HO-1信号通路相关蛋白的表达。建立心肌肥厚模型，将40只大鼠随机分为假手术组、模型组、葛根素组和葛根素+Nrf2抑制剂（ML385）组，每组10只。测量超声心动图、血流动力学参数和心肌肥厚参数。通过苏木精-伊红（HE）染色和Masson染色观察心肌组织的组织病理学变化。采用生化方法、酶联免疫吸附测定（ELISA）和荧光染色检测炎症因子和铁死亡相关指标。采用免疫组织化学法检测铁死亡相关蛋白以及Nrf2/ARE/HO-1信号通路相关蛋白的表达。细胞实验表明，葛根素干预可显著增强肥厚型心肌细胞的活力，减小其表面积，恢复线粒体膜电位和Ca²⁺稳态。机制研究显示，葛根素促进Nrf2核转位，上调HO-1、溶质载体家族7成员11（SLC7A11）和谷胱甘肽过氧化物酶4（GPX4）的表达，并降低丙二醛（MDA）、活性氧（ROS）和铁水平。ML385可逆转这些保护作用。在动物实验中，葛根素改善了心肌肥厚大鼠的心脏功能，减轻了心肌肥厚和纤维化，抑制了炎症反应和铁死亡，并促进了核Nrf2转位和HO-1表达。然而，与ML385联合干预导致血流动力学恶化和铁死亡标志物水平反弹。综上所述，葛根素可能通过Nrf2/ARE/HO-1信号通路抑制心肌细胞铁死亡，从而改善心肌肥厚中的心肌收缩功能。
